AIM To investigate the pathogenic effect ofSEB and D-GalN on liver and the protection ofcyclosporin A, the relationship between hepaticapoptosis and necrosis and the possiblemechanism of acute hepatic necrosis.METHODS...AIM To investigate the pathogenic effect ofSEB and D-GalN on liver and the protection ofcyclosporin A, the relationship between hepaticapoptosis and necrosis and the possiblemechanism of acute hepatic necrosis.METHODS After staphylococcal enterotoxin B(SEB ) mixed with D--galactosamine (D-GaiN )were injected intraperitoneally into Balb/c miceand those previously treated with cyclosporin A,blood samples were collected and livers wereisolated at 2, 6, 12 and 24 h. Patterns othepatocellular death were studiedmorphologically and biochemically, circulatingcytokines (TNF-a, IFN--y ) and mice mortalitywithin 24h was assessed.RESU’LTS The SEB could induce the typicalapoptotic changes of hepatocytes, the D-GaiNcould induce hepatocytes apoptosis anddegeneration at the same time, and the micehaving received the SEB + D-GaiN injectionsdeveloped apoptosis at 2 and 6 h, but after 12 hhepatocytes were characterized by severein jury, whereas all the examinations in thecyclosporin A treated mice were normal.CONCLUSION Hepatic cell apoptosis might berelated to necrosis, and massive hepatocyteapoptosis is likely the initiating step of acutehepatic necrosis in mice. The effects induced bySEB and D--GaiN on hepatocytes might bemediated by T cells, and could be prevented bycyclosporin A.展开更多
Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migra...Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.展开更多
BACKGROUND: At present, it has been confirmed that immunological rejection exists in the cell transplantation in brain tissue, the effects of immunosuppressant on the immunological rejection and the survival of grafts...BACKGROUND: At present, it has been confirmed that immunological rejection exists in the cell transplantation in brain tissue, the effects of immunosuppressant on the immunological rejection and the survival of grafts in brain cell transplantation are worthy being investigated further. OBJECTIVE: To observe the immunological rejection after transgeneic cell transplantation in treating cerebral hemorrhage in rats, and investigate the interventional effect of cyclosprin. DESIGN: A randomized controlled study. SETTINGS: Second Affiliated Hospital of Xuzhou Medical College; First Affiliated Hospital of Nanjing Medical University. MATERIALS: Thirty-five healthy clean-degree SD rats of 6-8 weeks old were used, weighing 200-250 g, either male or female; The FACSort flow cytometer (American BD Company) and NYD-1000 image analytical system were used. The rat-anti-rat CD4 monoclonal antibody, rat-anti-rat CD8 monoclonal antibody, and rat-anti-rat MHC Ⅱ antigen monoclonal antibody were purchased from Santa Cruz Company; SP and DAB kits were purchased from Beijing Zhongshan Bio-engineering Company. XSP-8C2 light microscope was the product of Shanghai Zousun Optical Instrument, Co.,Ltd, and KYKY-3800B electron microscope was the product of China KYKY Technology Development Co.,Ltd. METHODS: The experiments were carried out in the animal experimental center of Nanjing Medical University from April to July in 2003. ① Model establishment: The rats were anesthetized, and then the coordinates of left internal capsule were identified, and the needle was withdrawn after 120 μL blood was injected into the internal capsule. Adenoviruses were taken as the carriers, after the astrocytes were successfully transfected by nerve growth factor(NGF) gene, 0.2 mL cell suspension was injected into the sites of cerebral hemorrhage. Thirty successfully established rat models were randomly divided into cyclosporin A group (n=18) and control group, the rats were treated with intraperitoneal injection of cyclosporin A (10 mg/kg per day) intraperitoneal injection of saline of the same dosage from the 1st day after transplantation, once a day for 7 days continuously. ② CD4+ and CD8+ detection: The CD4+ and CD8+ T lymphocytes in caudal vein were counted with flow cytometer at 15 days after treatment. ③ Morphological observation in the transplanted sites: The rats were killed and then brain tissues were taken out, the transplanted sites and the structure of the normal brain tissue around the transplanted sites were observed with light and electron microscopes. ④ Detections of the infiltration of T lymphocyte subsets and expression of major histocompatibility complex (MHC) Ⅱ antigen in the transplanted sites: The image analysis of immunohistochemical sections was performed with the image analytical system, and the integral optical density (IOD) was taken as the statistical value to observe the infiltration of T lymphocyte subsets and expression of MHCⅡ antigen in the transplanted sites, and the normal brain tissue around the transplanted sites were taken as controls. MAIN OUTCOME MEASURES: ① Countings of CD4+ and CD8+ in peripheral blood; ②Results of the morphological observation in the transplanted sites; ③ Infiltration of T lymphocyte subsets and expression of MHC Ⅱ antigen in the transplanted sites. RESULTS: Totally 35 rats were used, and 30 were successfully made into models, 5 died during the treatment, the other 25 were involved in the analysis of results. ① Results of CD4+ and CD8+ T lymphocytes in peripheral blood: The percentages of CD4+ and CD8+ T lymphocytes in the cyclosporin A group were (29.20±3.97)% and (20.65±2.02)%, respectively, which were obviously lower than those in the control group [(47.39±3.01)%, (28.30±2.36)%, t=4.983, 4.012, P < 0.05], and the CD4+/CD8+ ratio was obviously lower than that in the control group (1.41±0.86, 1.68±0.69, t=3. 871, P < 0.05). ② Morphological results in the transplanted sites: Under optical and electron microscopes, the survival region of the transplant was round, and it had an unobvious migration region with the normal brain tissues, the grafts had normal cellular form. Infiltrations of lymphocytes and monocytes were observed in both groups, and mainly located in the transplanted sites, and the expression of lymphocytes in the cyclosporin A group was markedly lower than that in the control group, and no above-mentioned changes were observed in the normal brain tissue around the transplanted sites. ③ Results of CD4+ and CD8+ T lymphocytes and expression of MHC Ⅱ antigen in the transplanted sites: The CD4+ and CD8+ T lymphocytes and expression of MHC Ⅱ antigen in the transplanted sites were observed in both groups. The IOD of CD4+ and CD8+ antigen positive cells in the cyclosporin A group were obviously lower than those in the control group (1.85±0.38, 1.44±0.33; 3.33±0.37, 2.64±0.56, t=4.122, 4.434, P < 0.05), and the IOD of MHC Ⅱ antigen positive cells was markedly lower than that in the control group (0.76±0.22, 0.98±0.24, t=3.885, P < 0.05). CONCLUSION: ① There is immunological rejection in brain tissue after the transplantation of NSC transgeneic glial cells. ② The immunosuppressant of cyclosporin A can reduce the immunological rejection after the cell transplantation.展开更多
Purpose: Drugs that modify the production of cytokines may affect fracture healing.The immunosuppressive drug cyclosporin A is widely used to modify the immune response in transplantations and in treatment of rheumato...Purpose: Drugs that modify the production of cytokines may affect fracture healing.The immunosuppressive drug cyclosporin A is widely used to modify the immune response in transplantations and in treatment of rheumatoid disorders. We wanted to analyze the effect of cyclosporin A on fracture healing and on the development of trauma induced osteopenia.Methods: Experimental tibia fractures were stabilised with intramedullary pins in 26 rabbits. The animals were given 5mg/kg/day of cyclosporin A or placebo for 5 weeks. Bone mineral content, callus volume and biomechanical testing were performed on both tibias and femurs.Results: At 5 weeks cyclosporin A treatment resulted in increased bone mineral content and increased callus volume of the fractured bone. The femora on the operated side had significantly lower bone mineral content compared to the non-operated side. This trauma induced osteopenia was unaffected by cyclosporin A treatment. Failure torque and stiffness of the tibia and femora were similar in both groups.Interpretation: Cyclosporin A stimulates bone formation in fracture repair. The mechanism is unclear, but a direct or cytokine mediated effect on bone forming cells, or enhanced bone induction resulting in increased bone formation, is possible.展开更多
<Abstrat>The proliferation of mesangial cells on cyclosporin (CsA) test mediumwas studied by MTT assay and TNF-Q in cultured supernatant was examined byusing ELISA. The results showed that cyclosporin A signific...<Abstrat>The proliferation of mesangial cells on cyclosporin (CsA) test mediumwas studied by MTT assay and TNF-Q in cultured supernatant was examined byusing ELISA. The results showed that cyclosporin A significantly inhibited theproliferation of mesangial cells at the concentration between 0. 25 - 15 μg/ml(IC50 1μg/ml). This action appeared to be dose-dependent. Release of TNF-αfrom mesangial cells stimulated by LPS was also dose-dependently suppressed. Itis suggested that cyclosporin A play an important role in antiproliferation mecha-nism of mesangial cells in vitro.展开更多
AIM: To investigate the interaction between castanospermine and cyclosporin A(Cs A) and to provide an explanation for it.METHODS: The alkaloid castanospermine was prepared from the seeds of Castanospermum austral cons...AIM: To investigate the interaction between castanospermine and cyclosporin A(Cs A) and to provide an explanation for it.METHODS: The alkaloid castanospermine was prepared from the seeds of Castanospermum austral consistently achieving purity. Rat heterotopic cardiac transplantation and mixed lymphocyte reactivity were done using genetically inbred strains of PVG(donor) and DA(recipient). For the mixed lymphocyte reaction stimulator cells were irradiated with 3000 rads using a linear accelerator. Cyclosporin A was administered by gavage and venous blood collected 2 h later(C_2). The blood levels of Cs A(Neoral) were measured by immunoassay which consisted of a homogeneous enzyme assay(EMIT) on Cobas Mira. Statistical analyses of interactions were done by an acceleratedfailure time model with Weibull distribution for allograft survival and logistic regression for the mixed lymphocyte reactivity.RESULTS: Castanospermine prolonged transplant survival times as a function of dose even at relatively low doses. Cyclosporin A also prolonged transplant survival times as a function of dose particularly at doses above 2 mg/kg. There were synergistic interactions between castanospermine and Cs A in the prolongation of cardiac allograft survival for dose ranges of Cs A by castanospermine of(0 to 2) mg/kg by(0 to 200) mg/kg(HR = 0.986; 95%CI: 0.981-0.992; P < 0.001) and(0 to 3) mg/kg by(0 to 100) mg/kg(HR = 0.986; 95%CI: 0.981-0.992; P < 0.001) respectively. The addition of castanospermine did not significantly increase the levels of cyclosporin A on day 3 or day 6 for all doses of Cs A. On the contrary, cessation of castanospermine in the presence of Cs A at 2 mg/kg significantly increased the Cs A level(P = 0.002). Castanospermine inhibited mixed lymphocyte reactivity in a dose dependent manner but without synergistic interaction. CONCLUSION: There is synergistic interaction between castanospermine and Cs A in rat cardiac transplantation. Neither the mixed lymphocyte reaction nor the metabolism of Cs A provides an explanation.展开更多
Objective: To explore the therapy to further elevate the efficacy of the treatment of chronic aplastic anemia (CAA). Methods: Forty-five patients with CCA were assigned into two groups, the 26 patients in the treated ...Objective: To explore the therapy to further elevate the efficacy of the treatment of chronic aplastic anemia (CAA). Methods: Forty-five patients with CCA were assigned into two groups, the 26 patients in the treated group were treated by Shengxuening (生血宁, a Chinese herbal preparation) and cyclosporin A (CsA), and the 19 patients in the control group were treated with androgen alone, with the therapeutic course lasting for over 3 months. Changes of peripheral blood picture, and the colony productivity of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte macrophage (CFU-GM) in bone marrow were observed before and after 3 months treatment. The amount of erythrocyte and platelet infusion, frequency of infection, condition of hemorrhage and relevant death were also observed. The follow-up study was conducted for over half a year.Results: The total effective rate in the treated group was 84.6%, which was significantly higher than that in the control group (52.6%, P<0.05). Levels of hemoglobin, reticulocyte, neutrophil and platelet increased after treatment in the treated group, as compared with those before treatment, with significant difference (P<0.05), and the colony productivity of BFU-E, CFU-E and CFU-GM in bone marrow also got significantly increased (P<0.01), and showed significant difference from those in the control group (P<0.05). Conclusion: Shengxuening-assisting CsA therapy is an effective measure for treatment of CAA.展开更多
This paper described the therapeutic efficacy of immunosuppressive (IS) treatment agents (ATG and CSA) and androgens in the treatment of childhood aplastic anemia (AA). The results showed that the overall curative rat...This paper described the therapeutic efficacy of immunosuppressive (IS) treatment agents (ATG and CSA) and androgens in the treatment of childhood aplastic anemia (AA). The results showed that the overall curative rate was 52.4% in the ATG therapy group (21 cases) and 58.3% in the CSA therapy group (12 cases) respectively. The effective rate of all patients (SAA and CAA) was 58.1% in the IS group (18/31) and 40.4% in the androgens group (42/104), P】0.05. But, in the childhood patients with SAA, the clinical effective rate was 68.4% in the IS group and 7.9% in the androgens group, P【0.01. The laboratory tests revealed that the majority of the AA patients displalyed abnormal immunological states: inversed CD4/ CD8 ratio and increased IL-2 activity. These abnormal immunological states could be normalized in several patients when clinical response was abtained following IS therapy with ATG and CSA.展开更多
Both calcium ionophore A23187 and endoplasmic reticulum Ca2+- ATPase inhibitor thapsigargin (Tg) could increase intracellular free calcium concentration and induce apoptosis in some cell lines. In the present study, w...Both calcium ionophore A23187 and endoplasmic reticulum Ca2+- ATPase inhibitor thapsigargin (Tg) could increase intracellular free calcium concentration and induce apoptosis in some cell lines. In the present study, we found that HL-60 cells treated with A23187 (1μg/ml) for 4 h or with Tg (0.5μg/ml) for 2 h showed typical characteristics of apoptosis. Pretreatment with nontoxic concentration of cyclosporin A (CsA) (1μg/ml) Could block these effects. Flow cytometric analysis of intracellular Ca2+ after staining with fluo-3 AM showed that CsA did not prevent the increase of intracellular calcium induced by A23187 or Tg, but it could maintain the high level of intracellular Ca2+ for a long time. These results suggest that CsA may prevent calcium- induced apoptosis by blocking the transportation of Ca2+ in HL-60cells.展开更多
Recently there has been a resurgence of interest in cyclic peptides due to their therapeutic advantages in terms of potency, permeability, proteolytic stability, and unique selectivity relative to traditional smaller ...Recently there has been a resurgence of interest in cyclic peptides due to their therapeutic advantages in terms of potency, permeability, proteolytic stability, and unique selectivity relative to traditional smaller drug molecules. Cyclosporin is a family of cyclic peptides widely used as autoimmune suppression agents. Cyclosporin analogs consist of eleven amino acids with the main difference lying at the side chain of its amino acid residues. In this study, a single step separation method was developed utilizing Supercritical Fluid Chromatography (SFC) to resolve five naturally occurring cyclosporin analogs (Cyclosporin A, B, C, D, and H) on a bare silica-packed column. The optimized method involved use of ethanol-modified carbon dioxide as mobile phase on a bare silica column at 80 °C and UV detection at 220 nm. Although column temperature and back pressure generally had insignificant effect on SFC separation, it was found in our study that increasing temperature and pressure greatly improved peak shape and resolution.展开更多
Injury to peripheral nerves during injections of therapeutic agents such as penicillin G potassium is common in developing countries. It has been shown that cyclosporin A, a powerful immunosuppressive agent, can retar...Injury to peripheral nerves during injections of therapeutic agents such as penicillin G potassium is common in developing countries. It has been shown that cyclosporin A, a powerful immunosuppressive agent, can retard Wallerian degeneration after peripheral nerve crush injury. However, few studies are reported on the effects of cyclosporin A on peripheral nerve drug injection injury. This study aimed to assess the time-dependent efficacy of cyclosporine-A as an immunosuppressant therapy in an experimental rat nerve injection injury model established by penicillin G potassium injection. The rats were randomly divided into three groups based on the length of time after nerve injury induced by cyclosporine-A administration(30 minutes, 8 or 24 hours). The compound muscle action potentials were recorded pre-injury, early post-injury(within 1 hour) and 4 weeks after injury and compared statistically. Tissue samples were taken from each animal for histological analysis. Compared to the control group, a significant improvement of the compound muscle action potential amplitude value was observed only when cyclosporine-A was administered within 30 minutes of the injection injury(P < 0.05); at 8 or 24 hours after cyclosporine-A administration, compound muscle action potential amplitude was not changed compared with the control group. Thus, early immunosuppressant drug therapy may be a good alternative neuroprotective therapy option in experimental nerve injection injury induced by penicillin G potassium injection.展开更多
AIM: To elucidate the metabolism and the effect of the cyclosporin A (CyA) as a representative immuno-suppressive drug used in transplantation in a partially hepatectomized rat model. METHODS: CyA was administered to ...AIM: To elucidate the metabolism and the effect of the cyclosporin A (CyA) as a representative immuno-suppressive drug used in transplantation in a partially hepatectomized rat model. METHODS: CyA was administered to rats that under- went a 70% hepatectomy. These rats were randomly assigned into three groups according to the dose of CyA administration as follows; (group 1) water, (group 2) 5 mg/kg CyA, (group 3) 10 mg/kg CyA. On post- operative days-1, 3, 7 and 14, the rats were killed to analyze the serum concentration of CyA, the liver re- generation ratio, biochemical or histological markers, and mRNA expression using reverse transcriptase-poly- merase chain reaction method to determine albumin and cytochrome p450 expression. RESULTS: The serum concentration of CyA in group 3 was significantly higher than group 2 during liver regeneration. CyA enhanced the liver regeneration in a dose dependent manner. The mRNA expression associ- ated with CyA metabolism was signifi cantly decreased on day 14, while preserving the albumin producing activity. CONCLUSION: These data indicate that the p-450 ac-tivity required to metabolize the CyA may be reduced during regeneration of the remnant liver after a hepa- tectomy, which may, therefore, be linked to diffi culty in controlling the optimal dose of CyA during early period of LDLT.展开更多
In this study, the effect of cyclosporin A (CsA) eye drop on keratoconjunctivitis Sicca (KCS) and its mechanism were studied. The KCS models were established by injecting Pertussis vaccine, complete freund’s adjuvant...In this study, the effect of cyclosporin A (CsA) eye drop on keratoconjunctivitis Sicca (KCS) and its mechanism were studied. The KCS models were established by injecting Pertussis vaccine, complete freund’s adjuvant (CFA) and antigen of conjunctiva from isotype mice. Then the KCS models were treated with cyclosporin A eye drop. Changes in breaking-up time (BUT), lacrimal secretion in 30 min and diversion in 24 h were measured. The percentage of beaker cells, the lymphocytic infiltration in conjunctiva were observed. The expression levels of Aquaporin-3 (AQP3) in conjunctiva epithelial cells, beaker cells and accessory lacrimal gland were immunohistochemically detected. The results showed that there were significant differences in BUT, the percentage of beaker cells, lacrimal secretion in 30 min, the lymphocytic infiltration and the expression of AQP3 between the experimental group and an control group. It was concluded that CsA eye drop exerts marked therapeutic effect on KCS by inhibiting T lymph cells, increasing the goblet cells and AQP3 expression in conjunctiva.展开更多
Purpose: To select effective drugs against cellular proliferation in the vitreous. Methods: Cyclosporin A (0. 125mg/l - 4.0mg/l) was added to cultures of human retinal pigment epithelial (RPE) cells with or without ma...Purpose: To select effective drugs against cellular proliferation in the vitreous. Methods: Cyclosporin A (0. 125mg/l - 4.0mg/l) was added to cultures of human retinal pigment epithelial (RPE) cells with or without macrophage - conditioned medium (MCM) . Proliferation rate of the cells was measured with (3H) - thymidine incorporation and liquid scintillation techniques on days 3 and 5.Results: Cyclosporin A at a dosages of 0. 125mg/l had a slight inhibition on human RPE cells proliferation (-3.8%-4.1%, P>0.05).Cyclosporin A at a dose ranging from 0.25mg/l to 4.0mg/l inhibited cellular proliferation effectively and in a dose - dependent manner (8.7% -95.1%, P<0.05 r=0.94, P<0.001) with its ID50 of 1.49mg/l. In the culture of RPE with MCM, the inhibition on day 5 was more effective than that on day 3. Conclusion : Cyclosporin A had an effective inhibition on human RPE cell proliferation and it may be of potential use clinically. Eye science 1998; 14 : 41 - 44.展开更多
To the Editor:Myelosuppression is one of the most common side effects of chemotherapy.Some patients would develop long-lasting aplastic anemia(AA)even after stopping chemotherapy for over three months,due to persisten...To the Editor:Myelosuppression is one of the most common side effects of chemotherapy.Some patients would develop long-lasting aplastic anemia(AA)even after stopping chemotherapy for over three months,due to persistent destruction of the bone marrow.As a thrombopoietin-receptor agonist,eltrombopag(EPAG)has been widely used in patients with thrombocytopenia from various origins,[1]including those with AA secondary to chemotherapy.However,both EPAG and cyclosporin A(CsA)have been reported to possibly induce the expansion of abnormal clones or controversially increase the risk of neoplasms.[2]Thus,the challenge was to treat severe cytopenia while preventing cancer relapse.Our study reported the efficacy and safety of EPAG with or without CsA for patients with AA secondary to chemotherapy,in an attempt to find solutions to these difficulties.展开更多
文摘AIM To investigate the pathogenic effect ofSEB and D-GalN on liver and the protection ofcyclosporin A, the relationship between hepaticapoptosis and necrosis and the possiblemechanism of acute hepatic necrosis.METHODS After staphylococcal enterotoxin B(SEB ) mixed with D--galactosamine (D-GaiN )were injected intraperitoneally into Balb/c miceand those previously treated with cyclosporin A,blood samples were collected and livers wereisolated at 2, 6, 12 and 24 h. Patterns othepatocellular death were studiedmorphologically and biochemically, circulatingcytokines (TNF-a, IFN--y ) and mice mortalitywithin 24h was assessed.RESU’LTS The SEB could induce the typicalapoptotic changes of hepatocytes, the D-GaiNcould induce hepatocytes apoptosis anddegeneration at the same time, and the micehaving received the SEB + D-GaiN injectionsdeveloped apoptosis at 2 and 6 h, but after 12 hhepatocytes were characterized by severein jury, whereas all the examinations in thecyclosporin A treated mice were normal.CONCLUSION Hepatic cell apoptosis might berelated to necrosis, and massive hepatocyteapoptosis is likely the initiating step of acutehepatic necrosis in mice. The effects induced bySEB and D--GaiN on hepatocytes might bemediated by T cells, and could be prevented bycyclosporin A.
文摘Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.
基金a grant from Ap-plied Basic Research Programof Jiangsu Department of Sci-ence and Technology, No.BS99062
文摘BACKGROUND: At present, it has been confirmed that immunological rejection exists in the cell transplantation in brain tissue, the effects of immunosuppressant on the immunological rejection and the survival of grafts in brain cell transplantation are worthy being investigated further. OBJECTIVE: To observe the immunological rejection after transgeneic cell transplantation in treating cerebral hemorrhage in rats, and investigate the interventional effect of cyclosprin. DESIGN: A randomized controlled study. SETTINGS: Second Affiliated Hospital of Xuzhou Medical College; First Affiliated Hospital of Nanjing Medical University. MATERIALS: Thirty-five healthy clean-degree SD rats of 6-8 weeks old were used, weighing 200-250 g, either male or female; The FACSort flow cytometer (American BD Company) and NYD-1000 image analytical system were used. The rat-anti-rat CD4 monoclonal antibody, rat-anti-rat CD8 monoclonal antibody, and rat-anti-rat MHC Ⅱ antigen monoclonal antibody were purchased from Santa Cruz Company; SP and DAB kits were purchased from Beijing Zhongshan Bio-engineering Company. XSP-8C2 light microscope was the product of Shanghai Zousun Optical Instrument, Co.,Ltd, and KYKY-3800B electron microscope was the product of China KYKY Technology Development Co.,Ltd. METHODS: The experiments were carried out in the animal experimental center of Nanjing Medical University from April to July in 2003. ① Model establishment: The rats were anesthetized, and then the coordinates of left internal capsule were identified, and the needle was withdrawn after 120 μL blood was injected into the internal capsule. Adenoviruses were taken as the carriers, after the astrocytes were successfully transfected by nerve growth factor(NGF) gene, 0.2 mL cell suspension was injected into the sites of cerebral hemorrhage. Thirty successfully established rat models were randomly divided into cyclosporin A group (n=18) and control group, the rats were treated with intraperitoneal injection of cyclosporin A (10 mg/kg per day) intraperitoneal injection of saline of the same dosage from the 1st day after transplantation, once a day for 7 days continuously. ② CD4+ and CD8+ detection: The CD4+ and CD8+ T lymphocytes in caudal vein were counted with flow cytometer at 15 days after treatment. ③ Morphological observation in the transplanted sites: The rats were killed and then brain tissues were taken out, the transplanted sites and the structure of the normal brain tissue around the transplanted sites were observed with light and electron microscopes. ④ Detections of the infiltration of T lymphocyte subsets and expression of major histocompatibility complex (MHC) Ⅱ antigen in the transplanted sites: The image analysis of immunohistochemical sections was performed with the image analytical system, and the integral optical density (IOD) was taken as the statistical value to observe the infiltration of T lymphocyte subsets and expression of MHCⅡ antigen in the transplanted sites, and the normal brain tissue around the transplanted sites were taken as controls. MAIN OUTCOME MEASURES: ① Countings of CD4+ and CD8+ in peripheral blood; ②Results of the morphological observation in the transplanted sites; ③ Infiltration of T lymphocyte subsets and expression of MHC Ⅱ antigen in the transplanted sites. RESULTS: Totally 35 rats were used, and 30 were successfully made into models, 5 died during the treatment, the other 25 were involved in the analysis of results. ① Results of CD4+ and CD8+ T lymphocytes in peripheral blood: The percentages of CD4+ and CD8+ T lymphocytes in the cyclosporin A group were (29.20±3.97)% and (20.65±2.02)%, respectively, which were obviously lower than those in the control group [(47.39±3.01)%, (28.30±2.36)%, t=4.983, 4.012, P < 0.05], and the CD4+/CD8+ ratio was obviously lower than that in the control group (1.41±0.86, 1.68±0.69, t=3. 871, P < 0.05). ② Morphological results in the transplanted sites: Under optical and electron microscopes, the survival region of the transplant was round, and it had an unobvious migration region with the normal brain tissues, the grafts had normal cellular form. Infiltrations of lymphocytes and monocytes were observed in both groups, and mainly located in the transplanted sites, and the expression of lymphocytes in the cyclosporin A group was markedly lower than that in the control group, and no above-mentioned changes were observed in the normal brain tissue around the transplanted sites. ③ Results of CD4+ and CD8+ T lymphocytes and expression of MHC Ⅱ antigen in the transplanted sites: The CD4+ and CD8+ T lymphocytes and expression of MHC Ⅱ antigen in the transplanted sites were observed in both groups. The IOD of CD4+ and CD8+ antigen positive cells in the cyclosporin A group were obviously lower than those in the control group (1.85±0.38, 1.44±0.33; 3.33±0.37, 2.64±0.56, t=4.122, 4.434, P < 0.05), and the IOD of MHC Ⅱ antigen positive cells was markedly lower than that in the control group (0.76±0.22, 0.98±0.24, t=3.885, P < 0.05). CONCLUSION: ① There is immunological rejection in brain tissue after the transplantation of NSC transgeneic glial cells. ② The immunosuppressant of cyclosporin A can reduce the immunological rejection after the cell transplantation.
文摘Purpose: Drugs that modify the production of cytokines may affect fracture healing.The immunosuppressive drug cyclosporin A is widely used to modify the immune response in transplantations and in treatment of rheumatoid disorders. We wanted to analyze the effect of cyclosporin A on fracture healing and on the development of trauma induced osteopenia.Methods: Experimental tibia fractures were stabilised with intramedullary pins in 26 rabbits. The animals were given 5mg/kg/day of cyclosporin A or placebo for 5 weeks. Bone mineral content, callus volume and biomechanical testing were performed on both tibias and femurs.Results: At 5 weeks cyclosporin A treatment resulted in increased bone mineral content and increased callus volume of the fractured bone. The femora on the operated side had significantly lower bone mineral content compared to the non-operated side. This trauma induced osteopenia was unaffected by cyclosporin A treatment. Failure torque and stiffness of the tibia and femora were similar in both groups.Interpretation: Cyclosporin A stimulates bone formation in fracture repair. The mechanism is unclear, but a direct or cytokine mediated effect on bone forming cells, or enhanced bone induction resulting in increased bone formation, is possible.
文摘<Abstrat>The proliferation of mesangial cells on cyclosporin (CsA) test mediumwas studied by MTT assay and TNF-Q in cultured supernatant was examined byusing ELISA. The results showed that cyclosporin A significantly inhibited theproliferation of mesangial cells at the concentration between 0. 25 - 15 μg/ml(IC50 1μg/ml). This action appeared to be dose-dependent. Release of TNF-αfrom mesangial cells stimulated by LPS was also dose-dependently suppressed. Itis suggested that cyclosporin A play an important role in antiproliferation mecha-nism of mesangial cells in vitro.
基金Supported by Kiriwina Investments Limited of Australia
文摘AIM: To investigate the interaction between castanospermine and cyclosporin A(Cs A) and to provide an explanation for it.METHODS: The alkaloid castanospermine was prepared from the seeds of Castanospermum austral consistently achieving purity. Rat heterotopic cardiac transplantation and mixed lymphocyte reactivity were done using genetically inbred strains of PVG(donor) and DA(recipient). For the mixed lymphocyte reaction stimulator cells were irradiated with 3000 rads using a linear accelerator. Cyclosporin A was administered by gavage and venous blood collected 2 h later(C_2). The blood levels of Cs A(Neoral) were measured by immunoassay which consisted of a homogeneous enzyme assay(EMIT) on Cobas Mira. Statistical analyses of interactions were done by an acceleratedfailure time model with Weibull distribution for allograft survival and logistic regression for the mixed lymphocyte reactivity.RESULTS: Castanospermine prolonged transplant survival times as a function of dose even at relatively low doses. Cyclosporin A also prolonged transplant survival times as a function of dose particularly at doses above 2 mg/kg. There were synergistic interactions between castanospermine and Cs A in the prolongation of cardiac allograft survival for dose ranges of Cs A by castanospermine of(0 to 2) mg/kg by(0 to 200) mg/kg(HR = 0.986; 95%CI: 0.981-0.992; P < 0.001) and(0 to 3) mg/kg by(0 to 100) mg/kg(HR = 0.986; 95%CI: 0.981-0.992; P < 0.001) respectively. The addition of castanospermine did not significantly increase the levels of cyclosporin A on day 3 or day 6 for all doses of Cs A. On the contrary, cessation of castanospermine in the presence of Cs A at 2 mg/kg significantly increased the Cs A level(P = 0.002). Castanospermine inhibited mixed lymphocyte reactivity in a dose dependent manner but without synergistic interaction. CONCLUSION: There is synergistic interaction between castanospermine and Cs A in rat cardiac transplantation. Neither the mixed lymphocyte reaction nor the metabolism of Cs A provides an explanation.
文摘Objective: To explore the therapy to further elevate the efficacy of the treatment of chronic aplastic anemia (CAA). Methods: Forty-five patients with CCA were assigned into two groups, the 26 patients in the treated group were treated by Shengxuening (生血宁, a Chinese herbal preparation) and cyclosporin A (CsA), and the 19 patients in the control group were treated with androgen alone, with the therapeutic course lasting for over 3 months. Changes of peripheral blood picture, and the colony productivity of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte macrophage (CFU-GM) in bone marrow were observed before and after 3 months treatment. The amount of erythrocyte and platelet infusion, frequency of infection, condition of hemorrhage and relevant death were also observed. The follow-up study was conducted for over half a year.Results: The total effective rate in the treated group was 84.6%, which was significantly higher than that in the control group (52.6%, P<0.05). Levels of hemoglobin, reticulocyte, neutrophil and platelet increased after treatment in the treated group, as compared with those before treatment, with significant difference (P<0.05), and the colony productivity of BFU-E, CFU-E and CFU-GM in bone marrow also got significantly increased (P<0.01), and showed significant difference from those in the control group (P<0.05). Conclusion: Shengxuening-assisting CsA therapy is an effective measure for treatment of CAA.
文摘This paper described the therapeutic efficacy of immunosuppressive (IS) treatment agents (ATG and CSA) and androgens in the treatment of childhood aplastic anemia (AA). The results showed that the overall curative rate was 52.4% in the ATG therapy group (21 cases) and 58.3% in the CSA therapy group (12 cases) respectively. The effective rate of all patients (SAA and CAA) was 58.1% in the IS group (18/31) and 40.4% in the androgens group (42/104), P】0.05. But, in the childhood patients with SAA, the clinical effective rate was 68.4% in the IS group and 7.9% in the androgens group, P【0.01. The laboratory tests revealed that the majority of the AA patients displalyed abnormal immunological states: inversed CD4/ CD8 ratio and increased IL-2 activity. These abnormal immunological states could be normalized in several patients when clinical response was abtained following IS therapy with ATG and CSA.
文摘Both calcium ionophore A23187 and endoplasmic reticulum Ca2+- ATPase inhibitor thapsigargin (Tg) could increase intracellular free calcium concentration and induce apoptosis in some cell lines. In the present study, we found that HL-60 cells treated with A23187 (1μg/ml) for 4 h or with Tg (0.5μg/ml) for 2 h showed typical characteristics of apoptosis. Pretreatment with nontoxic concentration of cyclosporin A (CsA) (1μg/ml) Could block these effects. Flow cytometric analysis of intracellular Ca2+ after staining with fluo-3 AM showed that CsA did not prevent the increase of intracellular calcium induced by A23187 or Tg, but it could maintain the high level of intracellular Ca2+ for a long time. These results suggest that CsA may prevent calcium- induced apoptosis by blocking the transportation of Ca2+ in HL-60cells.
文摘Recently there has been a resurgence of interest in cyclic peptides due to their therapeutic advantages in terms of potency, permeability, proteolytic stability, and unique selectivity relative to traditional smaller drug molecules. Cyclosporin is a family of cyclic peptides widely used as autoimmune suppression agents. Cyclosporin analogs consist of eleven amino acids with the main difference lying at the side chain of its amino acid residues. In this study, a single step separation method was developed utilizing Supercritical Fluid Chromatography (SFC) to resolve five naturally occurring cyclosporin analogs (Cyclosporin A, B, C, D, and H) on a bare silica-packed column. The optimized method involved use of ethanol-modified carbon dioxide as mobile phase on a bare silica column at 80 °C and UV detection at 220 nm. Although column temperature and back pressure generally had insignificant effect on SFC separation, it was found in our study that increasing temperature and pressure greatly improved peak shape and resolution.
文摘Injury to peripheral nerves during injections of therapeutic agents such as penicillin G potassium is common in developing countries. It has been shown that cyclosporin A, a powerful immunosuppressive agent, can retard Wallerian degeneration after peripheral nerve crush injury. However, few studies are reported on the effects of cyclosporin A on peripheral nerve drug injection injury. This study aimed to assess the time-dependent efficacy of cyclosporine-A as an immunosuppressant therapy in an experimental rat nerve injection injury model established by penicillin G potassium injection. The rats were randomly divided into three groups based on the length of time after nerve injury induced by cyclosporine-A administration(30 minutes, 8 or 24 hours). The compound muscle action potentials were recorded pre-injury, early post-injury(within 1 hour) and 4 weeks after injury and compared statistically. Tissue samples were taken from each animal for histological analysis. Compared to the control group, a significant improvement of the compound muscle action potential amplitude value was observed only when cyclosporine-A was administered within 30 minutes of the injection injury(P < 0.05); at 8 or 24 hours after cyclosporine-A administration, compound muscle action potential amplitude was not changed compared with the control group. Thus, early immunosuppressant drug therapy may be a good alternative neuroprotective therapy option in experimental nerve injection injury induced by penicillin G potassium injection.
文摘AIM: To elucidate the metabolism and the effect of the cyclosporin A (CyA) as a representative immuno-suppressive drug used in transplantation in a partially hepatectomized rat model. METHODS: CyA was administered to rats that under- went a 70% hepatectomy. These rats were randomly assigned into three groups according to the dose of CyA administration as follows; (group 1) water, (group 2) 5 mg/kg CyA, (group 3) 10 mg/kg CyA. On post- operative days-1, 3, 7 and 14, the rats were killed to analyze the serum concentration of CyA, the liver re- generation ratio, biochemical or histological markers, and mRNA expression using reverse transcriptase-poly- merase chain reaction method to determine albumin and cytochrome p450 expression. RESULTS: The serum concentration of CyA in group 3 was significantly higher than group 2 during liver regeneration. CyA enhanced the liver regeneration in a dose dependent manner. The mRNA expression associ- ated with CyA metabolism was signifi cantly decreased on day 14, while preserving the albumin producing activity. CONCLUSION: These data indicate that the p-450 ac-tivity required to metabolize the CyA may be reduced during regeneration of the remnant liver after a hepa- tectomy, which may, therefore, be linked to diffi culty in controlling the optimal dose of CyA during early period of LDLT.
文摘In this study, the effect of cyclosporin A (CsA) eye drop on keratoconjunctivitis Sicca (KCS) and its mechanism were studied. The KCS models were established by injecting Pertussis vaccine, complete freund’s adjuvant (CFA) and antigen of conjunctiva from isotype mice. Then the KCS models were treated with cyclosporin A eye drop. Changes in breaking-up time (BUT), lacrimal secretion in 30 min and diversion in 24 h were measured. The percentage of beaker cells, the lymphocytic infiltration in conjunctiva were observed. The expression levels of Aquaporin-3 (AQP3) in conjunctiva epithelial cells, beaker cells and accessory lacrimal gland were immunohistochemically detected. The results showed that there were significant differences in BUT, the percentage of beaker cells, lacrimal secretion in 30 min, the lymphocytic infiltration and the expression of AQP3 between the experimental group and an control group. It was concluded that CsA eye drop exerts marked therapeutic effect on KCS by inhibiting T lymph cells, increasing the goblet cells and AQP3 expression in conjunctiva.
基金The project was supported by the national natural science foundation of China (No.39470742)
文摘Purpose: To select effective drugs against cellular proliferation in the vitreous. Methods: Cyclosporin A (0. 125mg/l - 4.0mg/l) was added to cultures of human retinal pigment epithelial (RPE) cells with or without macrophage - conditioned medium (MCM) . Proliferation rate of the cells was measured with (3H) - thymidine incorporation and liquid scintillation techniques on days 3 and 5.Results: Cyclosporin A at a dosages of 0. 125mg/l had a slight inhibition on human RPE cells proliferation (-3.8%-4.1%, P>0.05).Cyclosporin A at a dose ranging from 0.25mg/l to 4.0mg/l inhibited cellular proliferation effectively and in a dose - dependent manner (8.7% -95.1%, P<0.05 r=0.94, P<0.001) with its ID50 of 1.49mg/l. In the culture of RPE with MCM, the inhibition on day 5 was more effective than that on day 3. Conclusion : Cyclosporin A had an effective inhibition on human RPE cell proliferation and it may be of potential use clinically. Eye science 1998; 14 : 41 - 44.
基金Chinese Acadmy of Medical Sciences Innovation Fund for Medical Sciences(CIFMS 2021-1-I2M-003)PUMCH National High-Level Hospital Clinical Research Funding(Nos.2022-PUMCH-C-026,2022-PUMCH-D-002,2022-PUMCH-B-046)
文摘To the Editor:Myelosuppression is one of the most common side effects of chemotherapy.Some patients would develop long-lasting aplastic anemia(AA)even after stopping chemotherapy for over three months,due to persistent destruction of the bone marrow.As a thrombopoietin-receptor agonist,eltrombopag(EPAG)has been widely used in patients with thrombocytopenia from various origins,[1]including those with AA secondary to chemotherapy.However,both EPAG and cyclosporin A(CsA)have been reported to possibly induce the expansion of abnormal clones or controversially increase the risk of neoplasms.[2]Thus,the challenge was to treat severe cytopenia while preventing cancer relapse.Our study reported the efficacy and safety of EPAG with or without CsA for patients with AA secondary to chemotherapy,in an attempt to find solutions to these difficulties.
