Objective:To determine whether salvianolic acid B(Sal B)exerts protective effects on diabetic peripheral neuropathy by attenuating apoptosis and pyroptosis.Methods:RSC96 cells were primarily cultured with DMEM(5.6 mmo...Objective:To determine whether salvianolic acid B(Sal B)exerts protective effects on diabetic peripheral neuropathy by attenuating apoptosis and pyroptosis.Methods:RSC96 cells were primarily cultured with DMEM(5.6 mmol/L glucose),hyperglycemia(HG,125 mmol/L glucose)and Sal B(0.1,1,and 10μmol/L).Cells proliferation was measured by 3-(4,5-cimethylthiazol-2-yl)-2,5-dilphenyltetrazolium bromide assay.Reactive oxygen species(ROS)generation and apoptosis rate were detected by flow cytometry analysis.Western blot was performed to analyze the expressions of poly ADP-ribose polymerase(PARR),cleaved-caspase 3,cleavedcaspase 9,Bcl-2,Bax,NLRP3,ASC,and interleukin(IL)-1β.Results:Treatment with HG at a concentration of125 mmol/L attenuated cellular proliferation,while Sal B alleviated this injury(P<0.05).In addition,Sal B inhibited HG-induced ROS production and apoptosis rate(P<0.05).Furthermore,treatment with Sal B down-regulated HG-induced PARP,cleaved-caspase 3,cleaved-caspase 9,Bax,NLRP3,ASC,and IL-1βexpression,but mitigated HG-mediated down-regulation of Bcl-2 expression(P<0.05).Conclusion:Sal B may protect RSC96 cells against HG-induced cellular injury via the inhibition of apoptosis and pyroptosis activated by ROS.展开更多
Background Pancreatic β cells are susceptible to fatty acid-induced apoptosis. The 17β-estradiol （E2） protects pancreatic βcells from apoptosis, mediated by the estrogen receptor-a （ERa）. The mRNA level and pro...Background Pancreatic β cells are susceptible to fatty acid-induced apoptosis. The 17β-estradiol （E2） protects pancreatic βcells from apoptosis, mediated by the estrogen receptor-a （ERa）. The mRNA level and promoter activity of leukemia-related protein （LRP） 16 were significantly increased by E2 in E R-a and LRP 16 was a co-activator of ER-a. The aim of the study was to assess the effects of LRP16 on fatty acid-induced apoptosis in MIN6 cells. Methods Cells with over-expressing LRP16 were obtained by lipidosome transfection. Insulin content and glucose-stimulated insulin secretion （GSIS） were examined by radioimmunoassay. Western blotting was applied to detect protein expression. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） and flow cytometry. The forkhead boxO1 （FoxO1） subcellular localization was determined by immunocytochemical analysis. Results MIN6-LRP16 cells with overexpression of LRP16 were successfully established, and protein expression of LRP16 was 2.29-fold of that of control cells （MIN6-3.1, P 〈0.05）. Insulin content and GSIS in MIN6-LRP16 were substantially increased compared with those in control cells. When cells were stimulated with glucose, increased phosphorylation of extracellular signal-regulated kinase （ERK） 1/2 and serine-threonine kinase （Akt） were observed in MIN6-LRP16. When cells were under palmitate pressure, the TUNEL-positive rate in MIN6-LRP16 was （17.0±0.5）%, while it in MIN6-3.1 was （22.0±0.4）%. In palmitate-treated cells, attenuated Akt phosphorylation was observed, but the attenuation in Akt activity was partially restored in MIN6-LRP16 cells. Meanwhile, nuclear localization of FoxO1 in MIN6-LRP16 was apparently reduced compared with that in control cells. Conclusions LRP16 regulated insulin content and GSIS in MIN6 cells by ERK1/2 and Akt activated way. Meanwhile, LRP16 overexpression protected MIN6 cells from fatty acid-induced apoptosis by partially restoring Akt phosphorylation and inhibiting FoxO1 nuclear redistribution. Therefore, LRP16 played important roles not only in insulin content and GSIS but also in the antilipotoxic effect mediated by Akt/FoxO1 signaling.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.81673785 and 81874447)Natural Science Basic Research Program of Shaanxi(Nos.2020JQ-515 and 2020JZ-34)+1 种基金Shaanxi Province Administration of Traditional Chinese Medicine(No.2019-ZZ-JC048)Hospital Topic of the First Affiliated Hospital of Xi'an Jiaotong University(No.2018MS-32)。
文摘Objective:To determine whether salvianolic acid B(Sal B)exerts protective effects on diabetic peripheral neuropathy by attenuating apoptosis and pyroptosis.Methods:RSC96 cells were primarily cultured with DMEM(5.6 mmol/L glucose),hyperglycemia(HG,125 mmol/L glucose)and Sal B(0.1,1,and 10μmol/L).Cells proliferation was measured by 3-(4,5-cimethylthiazol-2-yl)-2,5-dilphenyltetrazolium bromide assay.Reactive oxygen species(ROS)generation and apoptosis rate were detected by flow cytometry analysis.Western blot was performed to analyze the expressions of poly ADP-ribose polymerase(PARR),cleaved-caspase 3,cleavedcaspase 9,Bcl-2,Bax,NLRP3,ASC,and interleukin(IL)-1β.Results:Treatment with HG at a concentration of125 mmol/L attenuated cellular proliferation,while Sal B alleviated this injury(P<0.05).In addition,Sal B inhibited HG-induced ROS production and apoptosis rate(P<0.05).Furthermore,treatment with Sal B down-regulated HG-induced PARP,cleaved-caspase 3,cleaved-caspase 9,Bax,NLRP3,ASC,and IL-1βexpression,but mitigated HG-mediated down-regulation of Bcl-2 expression(P<0.05).Conclusion:Sal B may protect RSC96 cells against HG-induced cellular injury via the inhibition of apoptosis and pyroptosis activated by ROS.
基金This work was supported by a grant from the National Natural Science Foundation of China （No. 30771042）.
文摘Background Pancreatic β cells are susceptible to fatty acid-induced apoptosis. The 17β-estradiol （E2） protects pancreatic βcells from apoptosis, mediated by the estrogen receptor-a （ERa）. The mRNA level and promoter activity of leukemia-related protein （LRP） 16 were significantly increased by E2 in E R-a and LRP 16 was a co-activator of ER-a. The aim of the study was to assess the effects of LRP16 on fatty acid-induced apoptosis in MIN6 cells. Methods Cells with over-expressing LRP16 were obtained by lipidosome transfection. Insulin content and glucose-stimulated insulin secretion （GSIS） were examined by radioimmunoassay. Western blotting was applied to detect protein expression. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） and flow cytometry. The forkhead boxO1 （FoxO1） subcellular localization was determined by immunocytochemical analysis. Results MIN6-LRP16 cells with overexpression of LRP16 were successfully established, and protein expression of LRP16 was 2.29-fold of that of control cells （MIN6-3.1, P 〈0.05）. Insulin content and GSIS in MIN6-LRP16 were substantially increased compared with those in control cells. When cells were stimulated with glucose, increased phosphorylation of extracellular signal-regulated kinase （ERK） 1/2 and serine-threonine kinase （Akt） were observed in MIN6-LRP16. When cells were under palmitate pressure, the TUNEL-positive rate in MIN6-LRP16 was （17.0±0.5）%, while it in MIN6-3.1 was （22.0±0.4）%. In palmitate-treated cells, attenuated Akt phosphorylation was observed, but the attenuation in Akt activity was partially restored in MIN6-LRP16 cells. Meanwhile, nuclear localization of FoxO1 in MIN6-LRP16 was apparently reduced compared with that in control cells. Conclusions LRP16 regulated insulin content and GSIS in MIN6 cells by ERK1/2 and Akt activated way. Meanwhile, LRP16 overexpression protected MIN6 cells from fatty acid-induced apoptosis by partially restoring Akt phosphorylation and inhibiting FoxO1 nuclear redistribution. Therefore, LRP16 played important roles not only in insulin content and GSIS but also in the antilipotoxic effect mediated by Akt/FoxO1 signaling.
