Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis...Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family.In the present study,we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.Methods HK-2 cells were cultured,transfected with microRNA-181a inhibitor for 48 hours,and stimulated with 50 μmol/L cisplatin for 24 hours.MicroRNA-181a expression was analyzed by real time PCR,and cell apoptosis was detected by flow cytometry.Moreover,Bcl-2 and Bcl-2-associated X protein （Bax） expression were measured by Western blotting.Results MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor,compared with that in untransfectd cells （21.19±2.01 vs.38.87±1.97,P 〈0.05）.Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor.Compared with DDP treated cells alone,Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.Conclusion One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.展开更多
Objective:To explore the potential of metanephric mesenchymal cells(MMCs)for osteogenesis and naringin’s ability to enhance this process and its molecular mechanism.Methods:Porcine MMCs at 70 days of gestation were u...Objective:To explore the potential of metanephric mesenchymal cells(MMCs)for osteogenesis and naringin’s ability to enhance this process and its molecular mechanism.Methods:Porcine MMCs at 70 days of gestation were used as tool cells,cultured in osteogenic induction medium,identified by immunocytochemistry staining.Osteogenic potential of porcine MMCs and naringin’s ability to enhance this process was tested by detecting changes in cell viability,alkaline phosphatase(ALP)activity,the expression of runt-related transcription factor 2(Runx2),osteopontin(OPN)and osteocalcin(OCN),and the formation of mineralized nodules,and the application of the p38 signaling pathway inhibitor SB203580 vitiated the osteogenesis-promoting effect of naringin.Results:Immunocytochemical staining showed that the cells were Vimentin and Six2(+),E-cadherin and CK-18(−).Naringin can activate the p38 signaling pathway to enhance the osteogenesis of porcine MMCs by increasing cell viability,ALP activity,the expressions of Runx2,OPN and OCN,and the formation of mineralized nodules(P<0.05).The application of p38 signaling pathway inhibitor SB203580 vitiated the osteogenesis-promoting effect of naringin,manifested by decreased ALP activity,the expressions of Runx2,OPN and OCN,and the formation of mineralized nodules(P<0.05).Conclusion:Naringin,the active ingredient of Chinese herbal medicine Rhizoma Drynariae for nourishing Shen(Kidney)and strengthening bone,enhances the osteogenic differentiation of renal MMCs through the p38 signaling pathway.展开更多
Background Glucolipotoxicity might play an important role in the β cell decompensation stage during the development of obesity-associated type 2 diabetes.Tissue inhibitor of metalloproteinase-1 （TIMP-1） inhibits ma...Background Glucolipotoxicity might play an important role in the β cell decompensation stage during the development of obesity-associated type 2 diabetes.Tissue inhibitor of metalloproteinase-1 （TIMP-1） inhibits matrix metalloproteinase （MMP） activity and regulates proliferation and apoptosis of a variety of cell types,including pancreatic β-cells.In the present study,we investigated whether TIMP-1 counteracts glucolipotoxicity in the pancreatic β-cell line INS-1.Methods INS-1 cells were incubated in normal or high glucose,with or without palmitate （0.4 mmol/L）,in the presence of TIMP-1 or MMP inhibitor GM60001.In some experiments,cells were pretreated with phosphatidylinositol-3 （Pl-3） kinase inhibitor,LY294002 or wortmannin.The amount of dead INS-1 cells was determined by HO342 and propidium iodide staining.Akt phosphorylation was evaluated by Western blotting analysis to investigate a possible mechanism of TIMP-1＇s action.Results TIMP-1 protected INS-1 cells from glucolipotoxicity independent of MMP inhibition.TIMP-1 stimulated Akt phosphorylation.Inhibition of the PI-3 kinase pathway abolished the survival effect of TIMP-1.Conclusion TIMP-1 may counteract glucolipotoxicity induced β-cell death via a PI-3 kinase pathway.展开更多
基金This work was supported by the National Natural Science Foundation of China （No.61101218 and No.81102673）,the New Star Program of Beijing Science and Technology Commission （No.2011111） and the National Basic Research Program of China （No.2011 CB944004）.
文摘Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family.In the present study,we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.Methods HK-2 cells were cultured,transfected with microRNA-181a inhibitor for 48 hours,and stimulated with 50 μmol/L cisplatin for 24 hours.MicroRNA-181a expression was analyzed by real time PCR,and cell apoptosis was detected by flow cytometry.Moreover,Bcl-2 and Bcl-2-associated X protein （Bax） expression were measured by Western blotting.Results MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor,compared with that in untransfectd cells （21.19±2.01 vs.38.87±1.97,P 〈0.05）.Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor.Compared with DDP treated cells alone,Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.Conclusion One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.
基金Supported by the National Natural Science Foundation of China(Nos.82174115 and 81774027)。
文摘Objective:To explore the potential of metanephric mesenchymal cells(MMCs)for osteogenesis and naringin’s ability to enhance this process and its molecular mechanism.Methods:Porcine MMCs at 70 days of gestation were used as tool cells,cultured in osteogenic induction medium,identified by immunocytochemistry staining.Osteogenic potential of porcine MMCs and naringin’s ability to enhance this process was tested by detecting changes in cell viability,alkaline phosphatase(ALP)activity,the expression of runt-related transcription factor 2(Runx2),osteopontin(OPN)and osteocalcin(OCN),and the formation of mineralized nodules,and the application of the p38 signaling pathway inhibitor SB203580 vitiated the osteogenesis-promoting effect of naringin.Results:Immunocytochemical staining showed that the cells were Vimentin and Six2(+),E-cadherin and CK-18(−).Naringin can activate the p38 signaling pathway to enhance the osteogenesis of porcine MMCs by increasing cell viability,ALP activity,the expressions of Runx2,OPN and OCN,and the formation of mineralized nodules(P<0.05).The application of p38 signaling pathway inhibitor SB203580 vitiated the osteogenesis-promoting effect of naringin,manifested by decreased ALP activity,the expressions of Runx2,OPN and OCN,and the formation of mineralized nodules(P<0.05).Conclusion:Naringin,the active ingredient of Chinese herbal medicine Rhizoma Drynariae for nourishing Shen(Kidney)and strengthening bone,enhances the osteogenic differentiation of renal MMCs through the p38 signaling pathway.
文摘Background Glucolipotoxicity might play an important role in the β cell decompensation stage during the development of obesity-associated type 2 diabetes.Tissue inhibitor of metalloproteinase-1 （TIMP-1） inhibits matrix metalloproteinase （MMP） activity and regulates proliferation and apoptosis of a variety of cell types,including pancreatic β-cells.In the present study,we investigated whether TIMP-1 counteracts glucolipotoxicity in the pancreatic β-cell line INS-1.Methods INS-1 cells were incubated in normal or high glucose,with or without palmitate （0.4 mmol/L）,in the presence of TIMP-1 or MMP inhibitor GM60001.In some experiments,cells were pretreated with phosphatidylinositol-3 （Pl-3） kinase inhibitor,LY294002 or wortmannin.The amount of dead INS-1 cells was determined by HO342 and propidium iodide staining.Akt phosphorylation was evaluated by Western blotting analysis to investigate a possible mechanism of TIMP-1＇s action.Results TIMP-1 protected INS-1 cells from glucolipotoxicity independent of MMP inhibition.TIMP-1 stimulated Akt phosphorylation.Inhibition of the PI-3 kinase pathway abolished the survival effect of TIMP-1.Conclusion TIMP-1 may counteract glucolipotoxicity induced β-cell death via a PI-3 kinase pathway.
