BACKGROUND The importance of the neuronal microenvironment has been recently highlighted in gut region-specific diabetic enteric neuropathy. Regionally distinct thickening of endothelial basement membrane(BM) of intes...BACKGROUND The importance of the neuronal microenvironment has been recently highlighted in gut region-specific diabetic enteric neuropathy. Regionally distinct thickening of endothelial basement membrane(BM) of intestinal capillaries supplying the myenteric ganglia coincide with neuronal damage in different intestinal segments. Accelerated synthesis of matrix molecules and reduced degradation of matrix components may also contribute to the imbalance of extracellular matrix dynamics resulting in BM thickening. Among the matrix degrading proteinases, matrix metalloproteinase 9(MMP9) and its tissue inhibitor(TIMP1) are essential in regulating extracellular matrix remodelling.AIM To evaluate the intestinal segment-specific effects of diabetes and insulin replacement on ganglionic BM thickness, MMP9 and TIMP1 expression.METHODS Ten weeks after the onset of hyperglycaemia gut segments were taken from the duodenum and ileum of streptozotocin-induced diabetic, insulin-treated diabetic and sex-and age-matched control rats. The thickness of BM surrounding myenteric ganglia was measured by electron microscopic morphometry. Wholemount preparations of myenteric plexus were prepared from the different gut regions for MMP9/TIMP1 double-labelling fluorescent immunohistochemistry. Post-embedding immunogold electron microscopy was applied on ultrathin sections to evaluate the MMP9 and TIMP1 expression in myenteric ganglia and their microenvironment from different gut segments and conditions. The MMP9 and TIMP1 messenger ribonucleic acid(m RNA) level was measured by quantitative polymerase chain reaction.RESULTS Ten weeks after the onset of hyperglycaemia, the ganglionic BM was significantly thickened in the diabetic ileum, while it remained intact in the duodenum. The immediate insulin treatment prevented the diabetes-related thickening of the BM surrounding the ileal myenteric ganglia. Quantification of particle density showed an increasing tendency for MMP9 and a decreasing tendency for TIMP1 from the proximal to the distal small intestine under control conditions. In the diabetic ileum, the number of MMP9-indicating gold particles decreased in myenteric ganglia, endothelial cells of capillaries and intestinal smooth muscle cells, however, it remained unchanged in all duodenal compartments. The MMP9/TIMP1 ratio was also decreased in ileal ganglia only. However, a marked segment-specific induction was revealed in MMP9 and TIMP1 at the m RNA levels.CONCLUSION These findings support that the regional decrease in MMP9 expression in myenteric ganglia and their microenvironment may contribute to extracellular matrix accumulation, resulting in a region-specific thickening of ganglionic BM.展开更多
BACKGROUND: The high level of matrix metalloproteinase 9(MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fi broblasts and affect wound heali...BACKGROUND: The high level of matrix metalloproteinase 9(MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fi broblasts and affect wound healing is still unclear. The present study aimed to observe changes in the biological behaviors of rat dermal fi broblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot.METHODS: A cell model of skin f ibroblast with high expression of MMP9 was established by coculture of high glucose(22.0 mmol/L) and homocysteine(100 μmol/L). A control group was incubated with normal glucose(5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwell were used to detect cell proliferation, viability, collagen(hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically signif icant.RESULTS: The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group(7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively, P<0.01). The proportion of S-phase cells, proliferation index, cell viability, collagen(hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group(P<0.01).CONCLUSION: Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fi broblasts.展开更多
Dry eye disease(DED),primarily classified as multifactorial ocular surface disorder,afflicts tens of millions of individuals worldwide,adversely impacting their quality of life.Extensive research has been conducted on...Dry eye disease(DED),primarily classified as multifactorial ocular surface disorder,afflicts tens of millions of individuals worldwide,adversely impacting their quality of life.Extensive research has been conducted on tear film analysis over the past decades,offering a range of tests to evaluate its volume,health,and integrity.Yet,early diagnosis and effective treatment for DED continue to pose significant challenges in clinical settings.Nevertheless,by recognizing key phenomena in DED such as ocular surface inflammation,hyperosmolarity,and tear film instability,this article provides a comprehensive overview of both traditional and recently developed methods for diagnosing and monitoring DED.The information serves as a valuable resource not only for clinical diagnosis but also for further research into DED.展开更多
OBJECTIVE: To explore the effects of Taohong Siwu decoction(桃红四物汤, THSWD) on the extracellular matrix of endometrium in rats following drug-induced abortion.METHODS: Thirty-six pregnant female rats were administe...OBJECTIVE: To explore the effects of Taohong Siwu decoction(桃红四物汤, THSWD) on the extracellular matrix of endometrium in rats following drug-induced abortion.METHODS: Thirty-six pregnant female rats were administered mifepristone and misoprostol to induce abortion, and amounts of uterine bleeding were recorded. Pathological damage and collagen accumulation were detected by hematoxylin-eosin staining and Masson’s trichrome staining in uterus, respectively. Myeloperoxidase was evaluated by immunohistochemistry.The expression levels of fibronectin, laminin, matrix metalloproteinase 9(MMP-9), and tissue inhibitor of metalloproteinase 1(TIMP-1) were quantified using western blotting.RESULTS: THSWD could promote endometrial protection in rats following drug-induced abortion.The contents of cellulose and myeloperoxidase were significantly decreased in uterine tissue of THSWD-treated groups. Moreover, THSWD significantly decreased the expression levels of fibronectin, laminin, and TIMP-1. THSWD also significantly increased MMP-9 expression and the MMP-9/TIMP-1 ratio.CONCLUSION: THSWD plays a critical role in endometrial protection by reducing extracellular matrix deposition and uterine fibrosis. These effects may have been achieved by increasing MMP-9, reducing TIMP-1, and/or altering the ratio of MMP-9/TIMP-1.展开更多
Metastasis is the main cause of cancer-specific death in patients with prostate cancer(PCa).Acyl-coenzyme A synthetase long-chain family member 3(ACSL3)is involved in the metabolic reprogramming of multiple types of c...Metastasis is the main cause of cancer-specific death in patients with prostate cancer(PCa).Acyl-coenzyme A synthetase long-chain family member 3(ACSL3)is involved in the metabolic reprogramming of multiple types of cancer cells,but its role in PCa metastasis remains largely unknown.Here,we determined the effect of overexpression or small interfering RNA-mediated depletion of ACSL3 on the migratory and invasive abilities of human PCa cell lines.We also conducted phospho-protein microarray analysis to identify signaling pathway components affected by ACSL3 modulation.Overexpression of ACSL3 promoted the migration and invasion of PCa cells,whereas ACSL3 downregulation had the opposite effects.Mechanistically,phospho-protein analysis showed that ACSL3 regulated the phosphorylation of AKT and the expression of matrix metalloproteinase9.Our results support a potential role for ACSL3 in promoting the metastatic behavior of PCa,possibly via AKT/matrix metalloproteinase9 pathways.Thus,ACSL3 could be a novel target for the development of treatments for PCa.展开更多
基金European Union and the Hungarian Government in the framework,No.EFOP-3.6.1-16-2016-00008Hungarian NKFIH fund project,No.FK131789(to Bódi N)+1 种基金János Bolyai Research Scholarship of the Hungarian Academy of Sciences(to Bódi N)and New National Excellence Program of the Ministry for Innovation and Technology from the source of the National Research,Development and Innovation Fund,No.ÚNKP-20-5(to Bódi N).
文摘BACKGROUND The importance of the neuronal microenvironment has been recently highlighted in gut region-specific diabetic enteric neuropathy. Regionally distinct thickening of endothelial basement membrane(BM) of intestinal capillaries supplying the myenteric ganglia coincide with neuronal damage in different intestinal segments. Accelerated synthesis of matrix molecules and reduced degradation of matrix components may also contribute to the imbalance of extracellular matrix dynamics resulting in BM thickening. Among the matrix degrading proteinases, matrix metalloproteinase 9(MMP9) and its tissue inhibitor(TIMP1) are essential in regulating extracellular matrix remodelling.AIM To evaluate the intestinal segment-specific effects of diabetes and insulin replacement on ganglionic BM thickness, MMP9 and TIMP1 expression.METHODS Ten weeks after the onset of hyperglycaemia gut segments were taken from the duodenum and ileum of streptozotocin-induced diabetic, insulin-treated diabetic and sex-and age-matched control rats. The thickness of BM surrounding myenteric ganglia was measured by electron microscopic morphometry. Wholemount preparations of myenteric plexus were prepared from the different gut regions for MMP9/TIMP1 double-labelling fluorescent immunohistochemistry. Post-embedding immunogold electron microscopy was applied on ultrathin sections to evaluate the MMP9 and TIMP1 expression in myenteric ganglia and their microenvironment from different gut segments and conditions. The MMP9 and TIMP1 messenger ribonucleic acid(m RNA) level was measured by quantitative polymerase chain reaction.RESULTS Ten weeks after the onset of hyperglycaemia, the ganglionic BM was significantly thickened in the diabetic ileum, while it remained intact in the duodenum. The immediate insulin treatment prevented the diabetes-related thickening of the BM surrounding the ileal myenteric ganglia. Quantification of particle density showed an increasing tendency for MMP9 and a decreasing tendency for TIMP1 from the proximal to the distal small intestine under control conditions. In the diabetic ileum, the number of MMP9-indicating gold particles decreased in myenteric ganglia, endothelial cells of capillaries and intestinal smooth muscle cells, however, it remained unchanged in all duodenal compartments. The MMP9/TIMP1 ratio was also decreased in ileal ganglia only. However, a marked segment-specific induction was revealed in MMP9 and TIMP1 at the m RNA levels.CONCLUSION These findings support that the regional decrease in MMP9 expression in myenteric ganglia and their microenvironment may contribute to extracellular matrix accumulation, resulting in a region-specific thickening of ganglionic BM.
基金supported by grants from the National Natural Science Foundation of China(81070660)the Science and Technology Project Foundation of Guangdong Province(2008A030201012)+1 种基金Medical Science and Technology Research Foundation of Guangdong Province(A2012183)the Science and Technology Project Foundation of Guangdong Province(2009B091300128)
文摘BACKGROUND: The high level of matrix metalloproteinase 9(MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fi broblasts and affect wound healing is still unclear. The present study aimed to observe changes in the biological behaviors of rat dermal fi broblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot.METHODS: A cell model of skin f ibroblast with high expression of MMP9 was established by coculture of high glucose(22.0 mmol/L) and homocysteine(100 μmol/L). A control group was incubated with normal glucose(5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwell were used to detect cell proliferation, viability, collagen(hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically signif icant.RESULTS: The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group(7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively, P<0.01). The proportion of S-phase cells, proliferation index, cell viability, collagen(hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group(P<0.01).CONCLUSION: Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fi broblasts.
文摘Dry eye disease(DED),primarily classified as multifactorial ocular surface disorder,afflicts tens of millions of individuals worldwide,adversely impacting their quality of life.Extensive research has been conducted on tear film analysis over the past decades,offering a range of tests to evaluate its volume,health,and integrity.Yet,early diagnosis and effective treatment for DED continue to pose significant challenges in clinical settings.Nevertheless,by recognizing key phenomena in DED such as ocular surface inflammation,hyperosmolarity,and tear film instability,this article provides a comprehensive overview of both traditional and recently developed methods for diagnosing and monitoring DED.The information serves as a valuable resource not only for clinical diagnosis but also for further research into DED.
基金Supported by Natural Science Foundation-funded Projects: Regulation Effect of Taohong-Siwu Decoction on Alpha-granule Release of Platelet in Postpartum Blood Stasis, its Active Compounds and Action Pathway (No. 81473387)Mechanism Research of Taohong-Siwu Decoction in Regulating Brain Microvascular Angiogenesis of MACO Rats Based on Platelet Microparticles as a Plaletet-Brain Microvascular Cell Messenger (No. 81503291)Natural Science Foundation Projects of Anhui Colleges and Universities: Based on the Crosstalk of NLRP3-Caspase-1 and VEGF-Notch to Explore the Effect of THSWD Regulation of "Dispelling Stasis to Promote Regeneration" by the Interaction Between Pyroptosis and Angiogenesis After Cerebral Ischemia and Reperfusion (No. KJ2019A0467)。
文摘OBJECTIVE: To explore the effects of Taohong Siwu decoction(桃红四物汤, THSWD) on the extracellular matrix of endometrium in rats following drug-induced abortion.METHODS: Thirty-six pregnant female rats were administered mifepristone and misoprostol to induce abortion, and amounts of uterine bleeding were recorded. Pathological damage and collagen accumulation were detected by hematoxylin-eosin staining and Masson’s trichrome staining in uterus, respectively. Myeloperoxidase was evaluated by immunohistochemistry.The expression levels of fibronectin, laminin, matrix metalloproteinase 9(MMP-9), and tissue inhibitor of metalloproteinase 1(TIMP-1) were quantified using western blotting.RESULTS: THSWD could promote endometrial protection in rats following drug-induced abortion.The contents of cellulose and myeloperoxidase were significantly decreased in uterine tissue of THSWD-treated groups. Moreover, THSWD significantly decreased the expression levels of fibronectin, laminin, and TIMP-1. THSWD also significantly increased MMP-9 expression and the MMP-9/TIMP-1 ratio.CONCLUSION: THSWD plays a critical role in endometrial protection by reducing extracellular matrix deposition and uterine fibrosis. These effects may have been achieved by increasing MMP-9, reducing TIMP-1, and/or altering the ratio of MMP-9/TIMP-1.
基金Financial support This work was supported by the Pearl River S&T Nova Program of Guangzhou,China(no.201710010039)the Natural Science Foundation of Guangdong Province,China(no.2015A030313031 and 2017A030313898)+2 种基金the Science and Technology Program of Guangdong Province,China(no.2017A020215028)the Science and Technology Program of Guangzhou(no.201707010113)and the Basic Service Charge Young Teachers Cultivation Project of Sun Yat-sen University,China(no.17ykpy48).
文摘Metastasis is the main cause of cancer-specific death in patients with prostate cancer(PCa).Acyl-coenzyme A synthetase long-chain family member 3(ACSL3)is involved in the metabolic reprogramming of multiple types of cancer cells,but its role in PCa metastasis remains largely unknown.Here,we determined the effect of overexpression or small interfering RNA-mediated depletion of ACSL3 on the migratory and invasive abilities of human PCa cell lines.We also conducted phospho-protein microarray analysis to identify signaling pathway components affected by ACSL3 modulation.Overexpression of ACSL3 promoted the migration and invasion of PCa cells,whereas ACSL3 downregulation had the opposite effects.Mechanistically,phospho-protein analysis showed that ACSL3 regulated the phosphorylation of AKT and the expression of matrix metalloproteinase9.Our results support a potential role for ACSL3 in promoting the metastatic behavior of PCa,possibly via AKT/matrix metalloproteinase9 pathways.Thus,ACSL3 could be a novel target for the development of treatments for PCa.
