BACKGROUND Procalcitonin(Pct)is a common biomarker in clinical practice,especially in the era of coronavirus disease 2019(COVID-19)infection.Although it is frequently used for the diagnosis and prognostication of bact...BACKGROUND Procalcitonin(Pct)is a common biomarker in clinical practice,especially in the era of coronavirus disease 2019(COVID-19)infection.Although it is frequently used for the diagnosis and prognostication of bacterial infections or sepsis,it is also elevated in a few other conditions,including medullary thyroid carcinoma(MTC).CASE SUMMARY A 43-year-old female presented with moderately severe COVID-19 pneumonia in April 2021.She gradually recovered clinically;however,despite normalization of other inflammatory markers,Pct levels remained persistently elevated.Further workup identified the cause as left lobe MTC with locoregional metastasis.Calcitonin levels were high,and carcinoembryonic antigen levels were normal.The patient underwent total thyroidectomy and neck dissection,which was followed by another radical neck dissection due to residual disease.Currently,she is doing well,nearly having completed her course of external beam radiotherapy with no recurrence.Pct is well documented as a screening tool for MTC,especially because of its stable nature compared to calcitonin in the community settings.It is important to keep in mind the differential diagnosis of MTC in patients with persistently elevated Pct levels despite normal levels of other acute phase reactants.To the best of our knowledge,this is the first report from Asia of such an incidental diagnosis of MTC due to persistently elevated Pct levels in a patient with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection.CONCLUSION Persistently elevated Pct levels can occur in any pro-inflammatory state including infections,sepsis,or acute respiratory distress syndrome.In the current setting,SARS-CoV-2 infection is one such clinical scenario,and in rare situations of persistent elevation,MTC may need to be ruled out.展开更多
Objective The aim of this study was to investigate the effects of rearranged during transfection(RET)mutation on the expressions of calcitonin(CTn)and procalcitonin(PCT)in sporadic medullary thyroid carcinoma(SMTC).Me...Objective The aim of this study was to investigate the effects of rearranged during transfection(RET)mutation on the expressions of calcitonin(CTn)and procalcitonin(PCT)in sporadic medullary thyroid carcinoma(SMTC).Methods RET mutation was detected by polymerase chain reaction direct sequencing in 64 cases of SMTC,and the expression levels of CTn and PCT in SMTC tissues were detected using the immunohistochemical streptavidin-perosidase(SP)method.The effect of RET mutations on the expression of CTn and PCT along with its relationship with clinicopathological parameters were analyzed.Results The expression rates of CTn and PCT in SMTC tissues were 90.6%(58/64)and 67.2%(43/64),respectively.CTn and PCT expression were found to be associated with tumor size and lymph node metastasis(P<0.05)but not with gender,age,or tumor capsule invasion(P>0.05).There was a significant correlation between CTn and PCT expression(r=0.269,P=0.041),and the intensity of positive CTn expression was positively correlated with RET mutation(r=0.507,P=0.000).However,PCT expression was not associated with RET mutation(r=0.188,P=0.136).Conclusion High expression of CTn and PCT was associated with the progression of medullary carcinoma,and the intensity of CTn expression was associated with RET mutation.PCT may provide valuable information for the diagnosis and prognosis of SMTC.展开更多
Most migraine patients suffer from cutaneous allodynia; however, the underlying mechanisms are unclear. Calcitonin gene-related peptide(CGRP) plays an important role in the pathophysiology of migraine, and it is there...Most migraine patients suffer from cutaneous allodynia; however, the underlying mechanisms are unclear. Calcitonin gene-related peptide(CGRP) plays an important role in the pathophysiology of migraine, and it is therefore, a potential therapeutic target for treating the pain. In the present study, a rat model of conscious migraine, induced by repeated electrical stimulation of the superior sagittal sinus, was established and treated with electroacupuncture at Fengchi(GB20)(depth of 2–3 mm, frequency of 2/15 Hz, intensity of 0.5–1.0 m A, 15 minutes/day, for 7 consecutive days). Electroacupuncture at GB20 significantly alleviated the decrease in hind paw and facial withdrawal thresholds and significantly lessened the increase in the levels of CGRP in the trigeminal ganglion, trigeminal nucleus caudalis and ventroposterior medial thalamic nucleus in rats with migraine. No CGRP-positive cells were detected in the trigeminal nucleus caudalis or ventroposterior medial thalamic nucleus by immunofluorescence. Our findings suggest that electroacupuncture treatment ameliorates migraine pain and associated cutaneous allodynia by modulating the trigeminovascular system ascending pathway, at least in part by inhibiting CGRP expression in the trigeminal ganglion.展开更多
To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cultured wi...To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cultured with osteoblasts on ivory slices placed in 24 well plates . 24 h later, conditioned media containing CGRP and/or IL 1β were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL 1β significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL 1β in a dose dependent manner. It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts.展开更多
Dear editor,It is with great interest that we read the article“Relationship of calcitonin gene-related peptide with disease progression and prognosis of patients with severe traumatic brain injury”(Chen et al.,2018)...Dear editor,It is with great interest that we read the article“Relationship of calcitonin gene-related peptide with disease progression and prognosis of patients with severe traumatic brain injury”(Chen et al.,2018).In this study,the authors evaluated 121 patients who were divided into mild/moderate traumatic brain injury(TBI)(n=61),severe TBI(n=35)and control(n=25)groups,and measured serum levels of calcitonin gene-related peptide(CGRP)and serum endothelin-1(ET-1).They found that low levels of CGRP and high levels of ET-1 were associated with high mortality at 6 months.Identification of morphological abnormalities on CT scans is very important for evaluating patients with TBI because different diagnoses are made based on different imaging findings(Maas et al.,2005).展开更多
AIM:To analyze the impact of calcitonin gene-related peptide(CGRP)in mouse keratitis after Aspergillus fumigatus(A.fumigatus)infection.METHODS:C57 BL/6 mice were treated subconjunctivally with different concentrations...AIM:To analyze the impact of calcitonin gene-related peptide(CGRP)in mouse keratitis after Aspergillus fumigatus(A.fumigatus)infection.METHODS:C57 BL/6 mice were treated subconjunctivally with different concentrations of exogenous CGRP,and BALB/c mice were treated with CGRP8-37(a CGRP antagonist)before corneas were infected with A.fumigatus.The cornea was assessed under the slit-lamp and the clinical score was recorded.The mRNA levels of IL-1β,TNF-α,IL-6,and MIP-2 were detected by quantitative real-time polymerase chain reaction(PCR),while the protein level of IL-1βwas determined by Western blotting.In vitro,RAW264.7 cells were used to investigate NLRP3 and IL-1βexpression induced by A.fumigatus after the pretreatment of exogenous CGRP or CGRP8-37.Cytokines expression in RAW264.7 cells was evaluated by real-time PCR and Western blotting.RESULTS:Using exogenous CGRP resulted in downregulated synthesis of IL-1βand MIP-2 stimulated by A.fumigatus in C57 BL/6 mice keratitis,and the synthesis of IL-1β,MIP-2 and IL-6 was up-regulated in BALB/c mice corneas after the pretreatment with CGRP8-37.Pretreatment with exogenous CGRP and CGRP8-37 did not influence TNF-αmRNA levels either in BALB/c or C57 BL/6 mice keratitis.The levels of NLRP3 and IL-1βwere both reduced in A.fumigatus stimulated-macrophages after treatment with exogenous CGRP.And CGRP8-37 pretreatment would increase NLRP3 and IL-1βlevels.CONCLUSION:CGRP may alleviate the inflammatory reaction in mice keratitis after infection with A.fumigatus.The anti-inflammatory effect may be related to the inhibition of NLRP3 expression by CGRP.展开更多
The effect of calcitonin on the blood glucose level in glucose-tolerance test of children with the first degree obesity was studied. Initial blood glucose level was normal and glucose-tolerance test per os didn’t exp...The effect of calcitonin on the blood glucose level in glucose-tolerance test of children with the first degree obesity was studied. Initial blood glucose level was normal and glucose-tolerance test per os didn’t expose glucose tolerance impairment in children under control. Calcitonin didn’t change initial glucose concentration but evoked impairment of glucose tolerance. The girls with the first degree obesity were more sensitive to hyperglycemic effect of calcitonin than the boys. Sex specificities must be taken into consideration in calcitonin treatment. Besides one ought mean that the increasing secretion of calcitonin occurs in stress-reactions. In these conditions endogenous calcitonin can make the same effect on the regulation of carbohydrate metabolism as exogenous injections of hormone.展开更多
AIM: To investigate the expression and role of calcitonin gene-related peptide(CGRP) in the mouse models induced by Aspergillus fumigatus(A. fumigatus). METHODS: C57 BL/6 mice were randomized into a control group and ...AIM: To investigate the expression and role of calcitonin gene-related peptide(CGRP) in the mouse models induced by Aspergillus fumigatus(A. fumigatus). METHODS: C57 BL/6 mice were randomized into a control group and A. fumigatus keratitis group. The cornea photography was assessed under the slit lamp and the clinical score was recorded after infection. Western blot, real-time polymerase chain reaction(PCR) and immunohistofluorescence analysis were applied to detect CGRP expression in cornea of both groups. In vitro, tests were conducted with C57 BL/6 mice macrophages to investigate CGRP expression after interaction with A. fumigatus. Cytokines expression induced by exogenous CGRP and the antagonist CGRP8-37 in A. fumigatus-exposed macrophages was evaluated by real-time PCR and ELISA.RESULTS: The cornea expression of CGRP was significantly elevated in C57 BL/6 mice corneas and macrophages after A. fumigatus infection. After treatment with exogenous CGRP, the levels of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and IL-6 were reduced, and IL-10 level was increased in the A. fumigatus stimulatedmacrophages. However, IL-1β, TNF-α and IL-6 levels were upregulated after pretreatment of CGRP8-37. But the m RNA levels of MIP-2, TGF-β and IL-10 were not changed. CONCLUSION: This study provides evidence that A. fumigatus increased CGRP expression. CGRP may play a protective role against inflammation in A. fumigatus keratitis.展开更多
Objective To study the immunoreactivity of Calcitonin gene-related peptide (CGRP) in the facial nerve when Hemifacial Spasm is occurring. Methods The electrophysiological technique was used to explore abnormal muscle ...Objective To study the immunoreactivity of Calcitonin gene-related peptide (CGRP) in the facial nerve when Hemifacial Spasm is occurring. Methods The electrophysiological technique was used to explore abnormal muscle response (AMR) which was characteristic of Hemifacial Spasm.The animal models of Hemifacial Spasm in New Zealand white rabbits were established by compressing the main trunk of artificial demyelinated facial nerve with the temporal superficial artery. At 6 weeks after surgery, the facial nerves were taken from the experimental group and control one, the immunohistochemistry for CGRP using polyclonal antibody with ABC kit was performed in the facial nerves; at the same time, the observation for the facial nerves of light and transmission electron microscope was performed. Results The facial nerve demyelinated and the axons retrogressively changed, CGRP immunoreactive positive fibers were significantly detected in experimental groups; whereas this phenomenon was not found in control group. Conclusion CGRP can nutrien the injured facial nerve and plays an important role in the pathogenesis of Hemifacial Spasm.展开更多
BACKGROUND In clinical work,85%-90%of malignant thyroid diseases are papillary thyroid cancer(PTC);thus,clinicians neglect other types of thyroid cancer,such as medullary thyroid carcinoma(MTC).CASE SUMMARY We report ...BACKGROUND In clinical work,85%-90%of malignant thyroid diseases are papillary thyroid cancer(PTC);thus,clinicians neglect other types of thyroid cancer,such as medullary thyroid carcinoma(MTC).CASE SUMMARY We report a 53-year-old female patient with a preoperative calcitonin level of 345 pg/mL.There was no definitive diagnosis of MTC by preoperative fine-needle aspiration cytology or intraoperative frozen pathology,but the presence of PTC and MTC was confirmed by postoperative paraffin pathology.The patient underwent total thyroidectomy and bilateral central lymph node dissection.Close follow-up at 1.5 years after surgery revealed no signs of recurrence or metastasis.CONCLUSION The issue in clinical work-up regarding types of thyroid cancer provides a novel and challenging idea for the surgical treatment of MTC.In the absence of central lymph node metastasis,it is worth addressing whether patients with high calcitonin can undergo total thyroidectomy and bilateral central lymph node dissection without bilateral lateral neck lymph node dissection.展开更多
Calcitonin receptor-like receptor(CRLR)is a human protein,that produces a calcitonin gene-related peptide receptor(CGRP)when associates with human receptor activity-modifying protein-1(HRAMP1).CGRP is believed to be i...Calcitonin receptor-like receptor(CRLR)is a human protein,that produces a calcitonin gene-related peptide receptor(CGRP)when associates with human receptor activity-modifying protein-1(HRAMP1).CGRP is believed to be involved in triggering of migraine.Many strategies were employed to design antimigraine drug using various CGRP antagonist/ligands but most of them have failed due to their inability to reach the target“CGRP receptor”as they get metabolized before conferring their pharmacological action and exhibit toxic effect to the liver.The present study,evaluated the binding of 13 phyto-chemical compounds of“Abrus precatorius”identified through GCMS analysis against the protein CGRP using“Discovery Studio software”.The molecular docking study and ADME/T properties prediction were performed with the compounds using C-DOCKER module.The results showed that five lead compounds of A.precatorius may act as good inhibitors for migraine headache and the compounds can be re-designed and synthesized for better antimigraine activity.展开更多
Peptide analogs of salmon calcitonin (sCT) were synthesized by using Fmoc-based chemistry on MBHA resins Salmon calcitonin was modified by 1) cysteines at p0sitions 1 and 7 were replaced by valine and alanine respecti...Peptide analogs of salmon calcitonin (sCT) were synthesized by using Fmoc-based chemistry on MBHA resins Salmon calcitonin was modified by 1) cysteines at p0sitions 1 and 7 were replaced by valine and alanine respectively to result in open chain analogs. 2) the glycine at position 30 was replaced by alanine, D-alanine and sarcosine respectively, and 3) some residues were deleted besides the above two modifications. A modified two-step deprotection / cleavage procedure, in which a solvent of TFA / TMSBr / thioanisole / EDT / m-cresol combines with HF cleavage,was adopted in SPPS.展开更多
Secondary structure of [Val. Ala]sCT- an analog of salmon calcitonin (sCT) not containing an N-terminal disultide bridge. was investigated by circular dichroism (CD) and Fourier-transform intrared spectroscopy (FTIR) ...Secondary structure of [Val. Ala]sCT- an analog of salmon calcitonin (sCT) not containing an N-terminal disultide bridge. was investigated by circular dichroism (CD) and Fourier-transform intrared spectroscopy (FTIR) methods. Both CD and FTIR results show that the main contbrmational structure of [Val Ala’]sCT in aqueous solution is random coil structure. while in trifluorethanol (TFE) it displays a strong α-helical structure. The relationship between the biological activity and the conformational structure of [Val, Ala] sCT is als0 discussed.展开更多
Objective: The current study was designed to investigate the relationship between the pattern of methylation of calcitonin (CT) gene and the activity of DNA methyltranferase (MTase). Methods: The methylation rate of t...Objective: The current study was designed to investigate the relationship between the pattern of methylation of calcitonin (CT) gene and the activity of DNA methyltranferase (MTase). Methods: The methylation rate of the 5’ region of the CT gene (CTMR) and the activity of MTase in 6 solid tumor cell lines and 2 leukemia cell lines were determined. CTMR was detected by polymerase chain reaction (PCR) with inter and external references in combination with restriction endonuclease and laser scan technique. MTase activity was examined by isotope labeled microassay. Results: Both CTMR and MTase activity in all tumor cell lines were significantly higher than that of control cells. The increased MTase activity was relative to elevated CTMR. Conclusion: There is prevalence of hypermethylation of CT gene and elevated activity of MTase in malignant cells. The increased MTase activity is one of the possible reasons for CT gene hypermethylation in tumor cells.展开更多
It was investigated that the localization and distribution of calcitonin gene-relatedpeptide (CGRP) in the rabbit lung by ABC-GDN immunohistochemistry.The results demon-strated that CGRP immunoreactivity was located i...It was investigated that the localization and distribution of calcitonin gene-relatedpeptide (CGRP) in the rabbit lung by ABC-GDN immunohistochemistry.The results demon-strated that CGRP immunoreactivity was located in neuroendocrine cells (NEC_s) and neu-roepthelial bodies (NEB_s),including all airways from bronchi,bronchioles,and alveolar ducts.The nerve fibers in walls of the intrapulmonary bronchi and blood vessels also had CGRP-immunoreactivity.Adjacent sections immunostained with antibodies to CGRP and 5-HT provedthe coexistence of CGRP and 5-HT in NECs and NEBs.This finding suggested the possibilityof functional interaction between the peptide and amine.展开更多
Objective: To observe the inhibitory effect of calcitonin gene--related peptide (CGRP) on adriamycininduced acute cardiotoxicity. Methods: Primarily cultured rat myocardial cells were treated with 10-6 mol/Ladriamycin...Objective: To observe the inhibitory effect of calcitonin gene--related peptide (CGRP) on adriamycininduced acute cardiotoxicity. Methods: Primarily cultured rat myocardial cells were treated with 10-6 mol/Ladriamycin and 10-6mol/L adriamycin + 10 8mol/I. CGRP. Lactate dehydrogenase (LDH ) activity in the mediumand the contents of malondialdehyde (MDA ). calcium. and magnesium in the myocardial cells were assayed.Results: In the adriamycin group, LDH activity in medium and calcium, MDA contents in myocardial cells weresignificantly increased compared with those in control group, and magnesium content in the myocardial cells wassignificantly reduced. In the adriamycin group. there was a positive correlation between LDH activity in themedium and MDA content in the myocardial cells. Meanwhile, in the adriamycin + CGRP group,- CGRP mightsignificantly reduce the leakage of LDH from myocardial cells, lessen the increase in calcium and MDA contentsand prevent the loss of magnesium. Conclusion: CGRP may inhibit adriamycin induced acute cardiotoxicity byinhibiting lipid peroxidation, attenuating calcium overload, magnesium loss, and protecting enzyme activity.展开更多
Chemical synthesis and cloning of human calcitonin (hCT) cDNA. Methods and Results:The cDNA sequence coding for human calcitonin (hCT) was divided into 6 fragments, synthesized by ABI 391 DNA synthesis instrument and ...Chemical synthesis and cloning of human calcitonin (hCT) cDNA. Methods and Results:The cDNA sequence coding for human calcitonin (hCT) was divided into 6 fragments, synthesized by ABI 391 DNA synthesis instrument and cloned into PGEM7Z(+) vector following phosphorylation, annealing and ligation of the 6 fragments. The sequence of cloned hCT cDNA was proved to be the same as designed by double DNA sequencing. Conclusion: A reliable and forthright method of chemical synthesis and cloning of oligo-peplides cDNA (or gene) was provided in this study. This paper is a basic work for expression of hCT cDNA.展开更多
This work aimed to investigate the effects of calcitonin gene-related peptide(CGRP)-modified mesenchymal stem cells(MSCs)on vascular stenosis in carotid balloon-injured rats.The CGRP gene labeled with EGFP was transfe...This work aimed to investigate the effects of calcitonin gene-related peptide(CGRP)-modified mesenchymal stem cells(MSCs)on vascular stenosis in carotid balloon-injured rats.The CGRP gene labeled with EGFP was transfected into bone marrow MSCs,and CGRP protein expression in MSCs was confirmed by immunofluorescence assays.A rat carotid balloon injury model was established using a surgicalmethod.CGRP-modifiedMSCs were orthotopically transplanted into the injured area of the rats.At 28 days after cell transplantation,EGFP and CD31 expression was detected by immunofluorescence staining.Hematoxylin-eosin(H&E)staining was used to detect the intima/media area of the injured carotid artery stenosis site,and the expression of proliferating cell nuclear antigen(PCNA)was detected by immunohistochemistry.MSCs from rat bone marrow positively expressed CD29 and negatively expressed CD45.In vivo immunofluorescence staining showed that EGFP expression was significantly increased at the vascular injury site of rats transplanted with MSC_CGRP compared with the control group on the 28th day after cell transplantation and that the damaged vessels exhibited continuous CD31 expression.H&E staining showed that the vascular intimal proliferation area of rats transplanted with MSC_CGRP was significantly reduced compared with that of other groups.Furthermore,the immunohistochemistry results showed that PCNA expression in the endothelium of the MSC_CGRP group was lower than that of the other groups.Therefore,MSCs transfected with the CGRP gene can express the CGRP protein,and the implantation of CGRPmodified MSCs at the damaged site after carotid balloon-induced injury can reduce the neointimal area.展开更多
BACKGROUND: Activation of N-methyl-D-aspartate receptor(NMDAR)is a key link of exitotoxictiy at the phase of cerebral ischemic injury.Because NMDAR is a main way to mediate internal flow of Ca2+ among glutamic acid re...BACKGROUND: Activation of N-methyl-D-aspartate receptor(NMDAR)is a key link of exitotoxictiy at the phase of cerebral ischemic injury.Because NMDAR is a main way to mediate internal flow of Ca2+ among glutamic acid receptors,over-excitation can cause neuronal apoptosis.Calcitonin gene related peptide has a strongly biological activity.On one hand,it can play the protective effect through inhibiting the expression of NMDAR1 mRNA;on the other hand,it can play the protective effect through down-regulating the expression of NMDAR1 mRNA by exogenous calcitonin gene related peptide.OBJECTIVE: To observe the expression of NMDAR1 and the regulatory effect of calcitonin gene related peptide on the expression of NMDAR1 mRNA and protein in the cerebral cortex of rats with focal cerebral ischemia/reperfusion(I/R).DESIGN: Randomized controlled animal study.SETTING: China Medical University.MATERIALS: A total of 216 healthy male Wistar rats,general grade,weighing 250-280 g,were selected in this study.Twelve rats were randomly selected to regard as control group;meanwhile,other 204 rats were used to establish middle cerebral artery occlusion/reperfusion (MACO)models.The main reagents were detailed as follows:calcitonin gene related peptide (Sigma Company);calcitonin gene related peptide kit (Boster Company);antibodyⅠ,Ⅱand antibody β-actin Ⅰ,Ⅱ of NMDAR1 mRNA and chemiluminescence reagent (Santa Cruz Company,USA)METHODS:The experiment was carried out in the Laboratory of Neurobiology of China Medical University from August 2005 to June 2006. ①Right MCAO models of rats were established to cause focal ischemia and scored based on Zea Longa five-grade scale. If the scores were 1,2 and 3 after wakefulness,the MACO models were established successfully and involved in the experiment. A total of 120 rats with successful modeling were randomly divided into I/R group and administration group with 60 in each group.All rats in the both groups were observed at five time points,including 6,12,24,48 and 72 hours after reperfusion and after 2-hour ischemia,with 12 experimental animals at each time point. Six rats were prepared for detection of hybridization in situ,and the other 6 were used for Western blotting histochemical detection.Rats in the control group were opened their skin to separate common carotid artery and not treated with line and drugs. In addition,rats in the I/R group were treated with 1 mL saline at 2 hours after focal cerebral ischemia,and then,rats in the administration group were treated with 1 mL(1 g/L)calcitonin gene related peptide at 2 hours after focal cerebral ischemia.②The expression of NMDAR1 mRNA was detected with hybridization in situ at various time points; moreover,the expression of NMDAR1 protein was measured with Western blotting method at various time points.The results were analvzed with Metamoph imaging analytical system.MAIN OUTCOME MEASURES:The expression of NMDAR1 mRNA and its protein in cortical neurons of rats at various time points.RESULTS:A total of 84 rats were excluded because of non-symptoms,exanimation or death;and then,132 rats were involved in the final analysis.The expression of NMDAR1 mRNA and its protein in cortical neurons of rafs in the control group was 0.205±0.001 and 0.184±0.001,respectively;after I/R,expression of NMDAR1 mRNA and its protein was up-regulated,especially,expression of mRNA at 6,12,24,48 and 72 hours was 0.245±0.003,0.287 ± 0.004,0.354±0.008,0.284±0.002 and 0.217±0.006,respectively;moreover,expression of protein at 6,12,24,48 and 72 hours was 0.222±0.003,0.261±0.028,0.311±0.004,0.259±0.013 and 0.210±0.008,respectively.There was significant difference between the two groups(0.205±0.001,P＜0.01).The expression was up-related in the former 24 hours, reached peak at 24 hours,down-regulated, and decreased to the level of control group at 72 hours.Except 72 hours.the expression of NMDAR1 mRNA and its protein was lower in administration group than that in I/R group at other four time points. In addition.the expression of mRNA at 6,12,24,48 and 72 hours was 0.223±0.005,0.243±0.001,0.292±0.002,0.250±0.003 and 0.213±0.003,respectively;moreover,the expression of protein at 6,12.24,48 and 72 hours was 0.216±0.006,0.245±0.025,0.276±0.003,0.241±0.045 and 0.202±0.013, respectively.There was significant difference at various time points(P＜0.05).CONCLUSION:The expressions of NMDAR1 mRNA and its protein of peripheral cortical neurons are up-related in ischemic area after focal cerebral I/R.Meanwhile,exogenous calcitonin gene related peptide can protect cortical neurons through inhibiting expression of NMDAR1 mRNA and its protein after focal cerebral I/R.展开更多
Objective: To evaluate the effects of Xiongzhi Dilong decoction(XZDLD) and its wind medicine on calcitonin gene-related peptide(CGRP)-induced migraine and explore the mechanism through the CGRP/inducible nitric oxide ...Objective: To evaluate the effects of Xiongzhi Dilong decoction(XZDLD) and its wind medicine on calcitonin gene-related peptide(CGRP)-induced migraine and explore the mechanism through the CGRP/inducible nitric oxide synthase(iNOS) pathway.Methods: Rats were divided into control, model, XZDLD, XZDLD(external wind), XZDLD(internal wind),and olcegepant groups. CGRP was injected into the dura mater to induce a migraine. The frequency of head scratching, cage climbing, and facial grooming was observed. The pain threshold, the levels of CGRP,pituitary adenylate cyclase activating polypeptide(PACAP), substance P(SP), and the plasma protein extravasation(PPE) ratio were measured. The phosphorylation levels of p38, extracellular signalregulated kinase 1/2(ERK1/2), and expression of iNOS were detected by western blot.Results: Compared with the model group, the three modified XZDLD groups showed reduced frequency of head scratching and cage climbing in the first 30 min(all P <.05). Facial grooming frequency was reduced in XZDLD and XZDLD(external wind) groups(P =.0003 and P =.0131, respectively). External wind medicine played a more important role in increasing mechanical pain threshold than internal wind medicine. Moreover, compared with the model group, the three modified XZDLD groups demonstrated reduced plasma levels of CGRP and PACAP(all P <.05). No difference in the SP level was observed among the six groups. XZDLD reduced PPE ratio. XZDLD and XZDLD(external wind) groups suppressed the CGRP/iNOS pathway by inhibiting the p-p38/p38 ratio and the expression of iNOS. No difference in pERK1/2/ERK1/2 ratio was detected among the six groups.Conclusion: XZDLD increases pain threshold, downregulates the expression of CGRP and PACAP, and reduces PPE ratio by inhibiting the CGRP/iNOS pathway. External wind medicine is more effective than internal one on improving facial grooming and head scratching, increasing the mechanical pain threshold, and inhibiting the expression of iNOS.展开更多
文摘BACKGROUND Procalcitonin(Pct)is a common biomarker in clinical practice,especially in the era of coronavirus disease 2019(COVID-19)infection.Although it is frequently used for the diagnosis and prognostication of bacterial infections or sepsis,it is also elevated in a few other conditions,including medullary thyroid carcinoma(MTC).CASE SUMMARY A 43-year-old female presented with moderately severe COVID-19 pneumonia in April 2021.She gradually recovered clinically;however,despite normalization of other inflammatory markers,Pct levels remained persistently elevated.Further workup identified the cause as left lobe MTC with locoregional metastasis.Calcitonin levels were high,and carcinoembryonic antigen levels were normal.The patient underwent total thyroidectomy and neck dissection,which was followed by another radical neck dissection due to residual disease.Currently,she is doing well,nearly having completed her course of external beam radiotherapy with no recurrence.Pct is well documented as a screening tool for MTC,especially because of its stable nature compared to calcitonin in the community settings.It is important to keep in mind the differential diagnosis of MTC in patients with persistently elevated Pct levels despite normal levels of other acute phase reactants.To the best of our knowledge,this is the first report from Asia of such an incidental diagnosis of MTC due to persistently elevated Pct levels in a patient with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection.CONCLUSION Persistently elevated Pct levels can occur in any pro-inflammatory state including infections,sepsis,or acute respiratory distress syndrome.In the current setting,SARS-CoV-2 infection is one such clinical scenario,and in rare situations of persistent elevation,MTC may need to be ruled out.
基金Supported by a grant from the Gansu Provincial Funding for Health Research(No.GSWSKY2018-13).
文摘Objective The aim of this study was to investigate the effects of rearranged during transfection(RET)mutation on the expressions of calcitonin(CTn)and procalcitonin(PCT)in sporadic medullary thyroid carcinoma(SMTC).Methods RET mutation was detected by polymerase chain reaction direct sequencing in 64 cases of SMTC,and the expression levels of CTn and PCT in SMTC tissues were detected using the immunohistochemical streptavidin-perosidase(SP)method.The effect of RET mutations on the expression of CTn and PCT along with its relationship with clinicopathological parameters were analyzed.Results The expression rates of CTn and PCT in SMTC tissues were 90.6%(58/64)and 67.2%(43/64),respectively.CTn and PCT expression were found to be associated with tumor size and lymph node metastasis(P<0.05)but not with gender,age,or tumor capsule invasion(P>0.05).There was a significant correlation between CTn and PCT expression(r=0.269,P=0.041),and the intensity of positive CTn expression was positively correlated with RET mutation(r=0.507,P=0.000).However,PCT expression was not associated with RET mutation(r=0.188,P=0.136).Conclusion High expression of CTn and PCT was associated with the progression of medullary carcinoma,and the intensity of CTn expression was associated with RET mutation.PCT may provide valuable information for the diagnosis and prognosis of SMTC.
基金supported by the National Natural Science Foundation of China,No.81603683a grant from the National Basic Research Program of China,No.2014CB543203a grant from the Beijing Municipal Science&Technology Commission of China,No.Z171100001017033
文摘Most migraine patients suffer from cutaneous allodynia; however, the underlying mechanisms are unclear. Calcitonin gene-related peptide(CGRP) plays an important role in the pathophysiology of migraine, and it is therefore, a potential therapeutic target for treating the pain. In the present study, a rat model of conscious migraine, induced by repeated electrical stimulation of the superior sagittal sinus, was established and treated with electroacupuncture at Fengchi(GB20)(depth of 2–3 mm, frequency of 2/15 Hz, intensity of 0.5–1.0 m A, 15 minutes/day, for 7 consecutive days). Electroacupuncture at GB20 significantly alleviated the decrease in hind paw and facial withdrawal thresholds and significantly lessened the increase in the levels of CGRP in the trigeminal ganglion, trigeminal nucleus caudalis and ventroposterior medial thalamic nucleus in rats with migraine. No CGRP-positive cells were detected in the trigeminal nucleus caudalis or ventroposterior medial thalamic nucleus by immunofluorescence. Our findings suggest that electroacupuncture treatment ameliorates migraine pain and associated cutaneous allodynia by modulating the trigeminovascular system ascending pathway, at least in part by inhibiting CGRP expression in the trigeminal ganglion.
文摘To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cultured with osteoblasts on ivory slices placed in 24 well plates . 24 h later, conditioned media containing CGRP and/or IL 1β were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL 1β significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL 1β in a dose dependent manner. It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts.
文摘Dear editor,It is with great interest that we read the article“Relationship of calcitonin gene-related peptide with disease progression and prognosis of patients with severe traumatic brain injury”(Chen et al.,2018).In this study,the authors evaluated 121 patients who were divided into mild/moderate traumatic brain injury(TBI)(n=61),severe TBI(n=35)and control(n=25)groups,and measured serum levels of calcitonin gene-related peptide(CGRP)and serum endothelin-1(ET-1).They found that low levels of CGRP and high levels of ET-1 were associated with high mortality at 6 months.Identification of morphological abnormalities on CT scans is very important for evaluating patients with TBI because different diagnoses are made based on different imaging findings(Maas et al.,2005).
基金Supported by National Natural Science Foundation of China(No.81870632)the Youth National Natural Science Foundation of China(No.81800800+1 种基金No.81700800)the Natural Science Foundation of Shandong Province(No.ZR2017BH025)。
文摘AIM:To analyze the impact of calcitonin gene-related peptide(CGRP)in mouse keratitis after Aspergillus fumigatus(A.fumigatus)infection.METHODS:C57 BL/6 mice were treated subconjunctivally with different concentrations of exogenous CGRP,and BALB/c mice were treated with CGRP8-37(a CGRP antagonist)before corneas were infected with A.fumigatus.The cornea was assessed under the slit-lamp and the clinical score was recorded.The mRNA levels of IL-1β,TNF-α,IL-6,and MIP-2 were detected by quantitative real-time polymerase chain reaction(PCR),while the protein level of IL-1βwas determined by Western blotting.In vitro,RAW264.7 cells were used to investigate NLRP3 and IL-1βexpression induced by A.fumigatus after the pretreatment of exogenous CGRP or CGRP8-37.Cytokines expression in RAW264.7 cells was evaluated by real-time PCR and Western blotting.RESULTS:Using exogenous CGRP resulted in downregulated synthesis of IL-1βand MIP-2 stimulated by A.fumigatus in C57 BL/6 mice keratitis,and the synthesis of IL-1β,MIP-2 and IL-6 was up-regulated in BALB/c mice corneas after the pretreatment with CGRP8-37.Pretreatment with exogenous CGRP and CGRP8-37 did not influence TNF-αmRNA levels either in BALB/c or C57 BL/6 mice keratitis.The levels of NLRP3 and IL-1βwere both reduced in A.fumigatus stimulated-macrophages after treatment with exogenous CGRP.And CGRP8-37 pretreatment would increase NLRP3 and IL-1βlevels.CONCLUSION:CGRP may alleviate the inflammatory reaction in mice keratitis after infection with A.fumigatus.The anti-inflammatory effect may be related to the inhibition of NLRP3 expression by CGRP.
文摘The effect of calcitonin on the blood glucose level in glucose-tolerance test of children with the first degree obesity was studied. Initial blood glucose level was normal and glucose-tolerance test per os didn’t expose glucose tolerance impairment in children under control. Calcitonin didn’t change initial glucose concentration but evoked impairment of glucose tolerance. The girls with the first degree obesity were more sensitive to hyperglycemic effect of calcitonin than the boys. Sex specificities must be taken into consideration in calcitonin treatment. Besides one ought mean that the increasing secretion of calcitonin occurs in stress-reactions. In these conditions endogenous calcitonin can make the same effect on the regulation of carbohydrate metabolism as exogenous injections of hormone.
基金Supported by the National Natural Science Foundation of China (No.81470609 No.81700800+5 种基金 No.81870632 No.81800800)the Youth National Natural Science Foundation of China (No.81500695)Natural Science Foundation of Shandong Province (No.ZR2013HQ007 No.ZR2017MH008 No.ZR2017BH025)
文摘AIM: To investigate the expression and role of calcitonin gene-related peptide(CGRP) in the mouse models induced by Aspergillus fumigatus(A. fumigatus). METHODS: C57 BL/6 mice were randomized into a control group and A. fumigatus keratitis group. The cornea photography was assessed under the slit lamp and the clinical score was recorded after infection. Western blot, real-time polymerase chain reaction(PCR) and immunohistofluorescence analysis were applied to detect CGRP expression in cornea of both groups. In vitro, tests were conducted with C57 BL/6 mice macrophages to investigate CGRP expression after interaction with A. fumigatus. Cytokines expression induced by exogenous CGRP and the antagonist CGRP8-37 in A. fumigatus-exposed macrophages was evaluated by real-time PCR and ELISA.RESULTS: The cornea expression of CGRP was significantly elevated in C57 BL/6 mice corneas and macrophages after A. fumigatus infection. After treatment with exogenous CGRP, the levels of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and IL-6 were reduced, and IL-10 level was increased in the A. fumigatus stimulatedmacrophages. However, IL-1β, TNF-α and IL-6 levels were upregulated after pretreatment of CGRP8-37. But the m RNA levels of MIP-2, TGF-β and IL-10 were not changed. CONCLUSION: This study provides evidence that A. fumigatus increased CGRP expression. CGRP may play a protective role against inflammation in A. fumigatus keratitis.
文摘Objective To study the immunoreactivity of Calcitonin gene-related peptide (CGRP) in the facial nerve when Hemifacial Spasm is occurring. Methods The electrophysiological technique was used to explore abnormal muscle response (AMR) which was characteristic of Hemifacial Spasm.The animal models of Hemifacial Spasm in New Zealand white rabbits were established by compressing the main trunk of artificial demyelinated facial nerve with the temporal superficial artery. At 6 weeks after surgery, the facial nerves were taken from the experimental group and control one, the immunohistochemistry for CGRP using polyclonal antibody with ABC kit was performed in the facial nerves; at the same time, the observation for the facial nerves of light and transmission electron microscope was performed. Results The facial nerve demyelinated and the axons retrogressively changed, CGRP immunoreactive positive fibers were significantly detected in experimental groups; whereas this phenomenon was not found in control group. Conclusion CGRP can nutrien the injured facial nerve and plays an important role in the pathogenesis of Hemifacial Spasm.
基金Supported by National Natural Science Foundation of China,No.81860469Zunyi Science and Technology Bureau,China,ZunShi KeHe HZ(2019)No.85Honghuagang District Science and Technology Bureau of Zunyi City,China,ZunHong KeHe Shezi(2018)No.12.
文摘BACKGROUND In clinical work,85%-90%of malignant thyroid diseases are papillary thyroid cancer(PTC);thus,clinicians neglect other types of thyroid cancer,such as medullary thyroid carcinoma(MTC).CASE SUMMARY We report a 53-year-old female patient with a preoperative calcitonin level of 345 pg/mL.There was no definitive diagnosis of MTC by preoperative fine-needle aspiration cytology or intraoperative frozen pathology,but the presence of PTC and MTC was confirmed by postoperative paraffin pathology.The patient underwent total thyroidectomy and bilateral central lymph node dissection.Close follow-up at 1.5 years after surgery revealed no signs of recurrence or metastasis.CONCLUSION The issue in clinical work-up regarding types of thyroid cancer provides a novel and challenging idea for the surgical treatment of MTC.In the absence of central lymph node metastasis,it is worth addressing whether patients with high calcitonin can undergo total thyroidectomy and bilateral central lymph node dissection without bilateral lateral neck lymph node dissection.
文摘Calcitonin receptor-like receptor(CRLR)is a human protein,that produces a calcitonin gene-related peptide receptor(CGRP)when associates with human receptor activity-modifying protein-1(HRAMP1).CGRP is believed to be involved in triggering of migraine.Many strategies were employed to design antimigraine drug using various CGRP antagonist/ligands but most of them have failed due to their inability to reach the target“CGRP receptor”as they get metabolized before conferring their pharmacological action and exhibit toxic effect to the liver.The present study,evaluated the binding of 13 phyto-chemical compounds of“Abrus precatorius”identified through GCMS analysis against the protein CGRP using“Discovery Studio software”.The molecular docking study and ADME/T properties prediction were performed with the compounds using C-DOCKER module.The results showed that five lead compounds of A.precatorius may act as good inhibitors for migraine headache and the compounds can be re-designed and synthesized for better antimigraine activity.
文摘Peptide analogs of salmon calcitonin (sCT) were synthesized by using Fmoc-based chemistry on MBHA resins Salmon calcitonin was modified by 1) cysteines at p0sitions 1 and 7 were replaced by valine and alanine respectively to result in open chain analogs. 2) the glycine at position 30 was replaced by alanine, D-alanine and sarcosine respectively, and 3) some residues were deleted besides the above two modifications. A modified two-step deprotection / cleavage procedure, in which a solvent of TFA / TMSBr / thioanisole / EDT / m-cresol combines with HF cleavage,was adopted in SPPS.
文摘Secondary structure of [Val. Ala]sCT- an analog of salmon calcitonin (sCT) not containing an N-terminal disultide bridge. was investigated by circular dichroism (CD) and Fourier-transform intrared spectroscopy (FTIR) methods. Both CD and FTIR results show that the main contbrmational structure of [Val Ala’]sCT in aqueous solution is random coil structure. while in trifluorethanol (TFE) it displays a strong α-helical structure. The relationship between the biological activity and the conformational structure of [Val, Ala] sCT is als0 discussed.
基金grants from the Ministry of Public Health of China (94-1-250) and CMB.
文摘Objective: The current study was designed to investigate the relationship between the pattern of methylation of calcitonin (CT) gene and the activity of DNA methyltranferase (MTase). Methods: The methylation rate of the 5’ region of the CT gene (CTMR) and the activity of MTase in 6 solid tumor cell lines and 2 leukemia cell lines were determined. CTMR was detected by polymerase chain reaction (PCR) with inter and external references in combination with restriction endonuclease and laser scan technique. MTase activity was examined by isotope labeled microassay. Results: Both CTMR and MTase activity in all tumor cell lines were significantly higher than that of control cells. The increased MTase activity was relative to elevated CTMR. Conclusion: There is prevalence of hypermethylation of CT gene and elevated activity of MTase in malignant cells. The increased MTase activity is one of the possible reasons for CT gene hypermethylation in tumor cells.
文摘It was investigated that the localization and distribution of calcitonin gene-relatedpeptide (CGRP) in the rabbit lung by ABC-GDN immunohistochemistry.The results demon-strated that CGRP immunoreactivity was located in neuroendocrine cells (NEC_s) and neu-roepthelial bodies (NEB_s),including all airways from bronchi,bronchioles,and alveolar ducts.The nerve fibers in walls of the intrapulmonary bronchi and blood vessels also had CGRP-immunoreactivity.Adjacent sections immunostained with antibodies to CGRP and 5-HT provedthe coexistence of CGRP and 5-HT in NECs and NEBs.This finding suggested the possibilityof functional interaction between the peptide and amine.
文摘Objective: To observe the inhibitory effect of calcitonin gene--related peptide (CGRP) on adriamycininduced acute cardiotoxicity. Methods: Primarily cultured rat myocardial cells were treated with 10-6 mol/Ladriamycin and 10-6mol/L adriamycin + 10 8mol/I. CGRP. Lactate dehydrogenase (LDH ) activity in the mediumand the contents of malondialdehyde (MDA ). calcium. and magnesium in the myocardial cells were assayed.Results: In the adriamycin group, LDH activity in medium and calcium, MDA contents in myocardial cells weresignificantly increased compared with those in control group, and magnesium content in the myocardial cells wassignificantly reduced. In the adriamycin group. there was a positive correlation between LDH activity in themedium and MDA content in the myocardial cells. Meanwhile, in the adriamycin + CGRP group,- CGRP mightsignificantly reduce the leakage of LDH from myocardial cells, lessen the increase in calcium and MDA contentsand prevent the loss of magnesium. Conclusion: CGRP may inhibit adriamycin induced acute cardiotoxicity byinhibiting lipid peroxidation, attenuating calcium overload, magnesium loss, and protecting enzyme activity.
文摘Chemical synthesis and cloning of human calcitonin (hCT) cDNA. Methods and Results:The cDNA sequence coding for human calcitonin (hCT) was divided into 6 fragments, synthesized by ABI 391 DNA synthesis instrument and cloned into PGEM7Z(+) vector following phosphorylation, annealing and ligation of the 6 fragments. The sequence of cloned hCT cDNA was proved to be the same as designed by double DNA sequencing. Conclusion: A reliable and forthright method of chemical synthesis and cloning of oligo-peplides cDNA (or gene) was provided in this study. This paper is a basic work for expression of hCT cDNA.
基金This work was supported by the Technology Fund of Guizhou Province(Qian ke he LH zi[2014]7576).
文摘This work aimed to investigate the effects of calcitonin gene-related peptide(CGRP)-modified mesenchymal stem cells(MSCs)on vascular stenosis in carotid balloon-injured rats.The CGRP gene labeled with EGFP was transfected into bone marrow MSCs,and CGRP protein expression in MSCs was confirmed by immunofluorescence assays.A rat carotid balloon injury model was established using a surgicalmethod.CGRP-modifiedMSCs were orthotopically transplanted into the injured area of the rats.At 28 days after cell transplantation,EGFP and CD31 expression was detected by immunofluorescence staining.Hematoxylin-eosin(H&E)staining was used to detect the intima/media area of the injured carotid artery stenosis site,and the expression of proliferating cell nuclear antigen(PCNA)was detected by immunohistochemistry.MSCs from rat bone marrow positively expressed CD29 and negatively expressed CD45.In vivo immunofluorescence staining showed that EGFP expression was significantly increased at the vascular injury site of rats transplanted with MSC_CGRP compared with the control group on the 28th day after cell transplantation and that the damaged vessels exhibited continuous CD31 expression.H&E staining showed that the vascular intimal proliferation area of rats transplanted with MSC_CGRP was significantly reduced compared with that of other groups.Furthermore,the immunohistochemistry results showed that PCNA expression in the endothelium of the MSC_CGRP group was lower than that of the other groups.Therefore,MSCs transfected with the CGRP gene can express the CGRP protein,and the implantation of CGRPmodified MSCs at the damaged site after carotid balloon-induced injury can reduce the neointimal area.
文摘BACKGROUND: Activation of N-methyl-D-aspartate receptor(NMDAR)is a key link of exitotoxictiy at the phase of cerebral ischemic injury.Because NMDAR is a main way to mediate internal flow of Ca2+ among glutamic acid receptors,over-excitation can cause neuronal apoptosis.Calcitonin gene related peptide has a strongly biological activity.On one hand,it can play the protective effect through inhibiting the expression of NMDAR1 mRNA;on the other hand,it can play the protective effect through down-regulating the expression of NMDAR1 mRNA by exogenous calcitonin gene related peptide.OBJECTIVE: To observe the expression of NMDAR1 and the regulatory effect of calcitonin gene related peptide on the expression of NMDAR1 mRNA and protein in the cerebral cortex of rats with focal cerebral ischemia/reperfusion(I/R).DESIGN: Randomized controlled animal study.SETTING: China Medical University.MATERIALS: A total of 216 healthy male Wistar rats,general grade,weighing 250-280 g,were selected in this study.Twelve rats were randomly selected to regard as control group;meanwhile,other 204 rats were used to establish middle cerebral artery occlusion/reperfusion (MACO)models.The main reagents were detailed as follows:calcitonin gene related peptide (Sigma Company);calcitonin gene related peptide kit (Boster Company);antibodyⅠ,Ⅱand antibody β-actin Ⅰ,Ⅱ of NMDAR1 mRNA and chemiluminescence reagent (Santa Cruz Company,USA)METHODS:The experiment was carried out in the Laboratory of Neurobiology of China Medical University from August 2005 to June 2006. ①Right MCAO models of rats were established to cause focal ischemia and scored based on Zea Longa five-grade scale. If the scores were 1,2 and 3 after wakefulness,the MACO models were established successfully and involved in the experiment. A total of 120 rats with successful modeling were randomly divided into I/R group and administration group with 60 in each group.All rats in the both groups were observed at five time points,including 6,12,24,48 and 72 hours after reperfusion and after 2-hour ischemia,with 12 experimental animals at each time point. Six rats were prepared for detection of hybridization in situ,and the other 6 were used for Western blotting histochemical detection.Rats in the control group were opened their skin to separate common carotid artery and not treated with line and drugs. In addition,rats in the I/R group were treated with 1 mL saline at 2 hours after focal cerebral ischemia,and then,rats in the administration group were treated with 1 mL(1 g/L)calcitonin gene related peptide at 2 hours after focal cerebral ischemia.②The expression of NMDAR1 mRNA was detected with hybridization in situ at various time points; moreover,the expression of NMDAR1 protein was measured with Western blotting method at various time points.The results were analvzed with Metamoph imaging analytical system.MAIN OUTCOME MEASURES:The expression of NMDAR1 mRNA and its protein in cortical neurons of rats at various time points.RESULTS:A total of 84 rats were excluded because of non-symptoms,exanimation or death;and then,132 rats were involved in the final analysis.The expression of NMDAR1 mRNA and its protein in cortical neurons of rafs in the control group was 0.205±0.001 and 0.184±0.001,respectively;after I/R,expression of NMDAR1 mRNA and its protein was up-regulated,especially,expression of mRNA at 6,12,24,48 and 72 hours was 0.245±0.003,0.287 ± 0.004,0.354±0.008,0.284±0.002 and 0.217±0.006,respectively;moreover,expression of protein at 6,12,24,48 and 72 hours was 0.222±0.003,0.261±0.028,0.311±0.004,0.259±0.013 and 0.210±0.008,respectively.There was significant difference between the two groups(0.205±0.001,P＜0.01).The expression was up-related in the former 24 hours, reached peak at 24 hours,down-regulated, and decreased to the level of control group at 72 hours.Except 72 hours.the expression of NMDAR1 mRNA and its protein was lower in administration group than that in I/R group at other four time points. In addition.the expression of mRNA at 6,12,24,48 and 72 hours was 0.223±0.005,0.243±0.001,0.292±0.002,0.250±0.003 and 0.213±0.003,respectively;moreover,the expression of protein at 6,12.24,48 and 72 hours was 0.216±0.006,0.245±0.025,0.276±0.003,0.241±0.045 and 0.202±0.013, respectively.There was significant difference at various time points(P＜0.05).CONCLUSION:The expressions of NMDAR1 mRNA and its protein of peripheral cortical neurons are up-related in ischemic area after focal cerebral I/R.Meanwhile,exogenous calcitonin gene related peptide can protect cortical neurons through inhibiting expression of NMDAR1 mRNA and its protein after focal cerebral I/R.
基金funded by the National Natural Science Foundation of China (81873256)the National Key R&D Program of China “Study on Modernization of Traditional Chinese Medicine”(2019YFC1709702)。
文摘Objective: To evaluate the effects of Xiongzhi Dilong decoction(XZDLD) and its wind medicine on calcitonin gene-related peptide(CGRP)-induced migraine and explore the mechanism through the CGRP/inducible nitric oxide synthase(iNOS) pathway.Methods: Rats were divided into control, model, XZDLD, XZDLD(external wind), XZDLD(internal wind),and olcegepant groups. CGRP was injected into the dura mater to induce a migraine. The frequency of head scratching, cage climbing, and facial grooming was observed. The pain threshold, the levels of CGRP,pituitary adenylate cyclase activating polypeptide(PACAP), substance P(SP), and the plasma protein extravasation(PPE) ratio were measured. The phosphorylation levels of p38, extracellular signalregulated kinase 1/2(ERK1/2), and expression of iNOS were detected by western blot.Results: Compared with the model group, the three modified XZDLD groups showed reduced frequency of head scratching and cage climbing in the first 30 min(all P <.05). Facial grooming frequency was reduced in XZDLD and XZDLD(external wind) groups(P =.0003 and P =.0131, respectively). External wind medicine played a more important role in increasing mechanical pain threshold than internal wind medicine. Moreover, compared with the model group, the three modified XZDLD groups demonstrated reduced plasma levels of CGRP and PACAP(all P <.05). No difference in the SP level was observed among the six groups. XZDLD reduced PPE ratio. XZDLD and XZDLD(external wind) groups suppressed the CGRP/iNOS pathway by inhibiting the p-p38/p38 ratio and the expression of iNOS. No difference in pERK1/2/ERK1/2 ratio was detected among the six groups.Conclusion: XZDLD increases pain threshold, downregulates the expression of CGRP and PACAP, and reduces PPE ratio by inhibiting the CGRP/iNOS pathway. External wind medicine is more effective than internal one on improving facial grooming and head scratching, increasing the mechanical pain threshold, and inhibiting the expression of iNOS.
