J147对高糖诱导人神经母细胞瘤细胞损伤保护作用的研究  

Protective effect of J147 on high glucose-induced SH-SY5Y cell injury

作　　者：黄晓雪 葛文贤 张亚茹 庄耀 丁淑敏[1] 吕金鹏 柳丽[1] UANG Xiaorue;GE Wencian;ZHANG Yaru(School of Pharmacy,Changzhou University,Changzhou 213100,China)

机构地区：[1]常州大学药学院,213100 [2]常州市武进中医医院检验科

出　　处：《中国糖尿病杂志》2024年第11期839-848,共10页Chinese Journal of Diabetes

基　　金：国家自然科学基金(82103752)。

摘　　要：目的 探讨J147对高糖(HG)诱导人神经母细胞瘤(SH-SY5Y)细胞损伤的保护作用及机制。方法 建立HG诱导SH-SY5Y细胞损伤模型,并将SH-SY5Y细胞分为正常对照(Con)组、HG组、HG+J147 0.5μmol/L(HG+J147 0.5)组、HG+J147 1μmol/L(HG+J147 1)组、HG+J147 2μmol/L(HG+J147 2)组、HG+J147 2μmol/L+磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)抑制剂LY294002(LY)10μmol/L(HG+J147 2+LY)组、HG+J147 2μmol/L+细胞外信号调节激酶1/2(ERK1/2)抑制剂U0126(U0)5μmol/L(HG+J147 2+U0)组、HG+LY 10μmol/L(HG+LY)组、HG+U0 5μmol/L(HG+U0)组。MTS细胞增殖与毒性检测试剂盒检测细胞活力,乳酸脱氢酶(LDH)试剂盒检测LDH活性,显微镜观察SH-SY5Y细胞形态学变化,流式细胞术检测细胞凋亡,Western blot法检测凋亡相关蛋白[凋亡B淋巴细胞瘤2(Bcl-2)、Bcl-2相关蛋白X(Bax)]和信号通路相关蛋白[AKT、p-AKT、ERK1/2、p-ERK1/2、环腺苷酸反应结合蛋白(CREB)、p-CREB、脑源性神经营养因子(BDNF)]蛋白表达。结果 与Con组比较,HG组细胞活力、Bcl-2/Bax比值、p-AKT/AKT、p-ERK/ERK、p-CREB/CREB、BDNF蛋白表达降低(P<0.01),LDH活性、细胞凋亡率升高(P<0.01)。与HG组比较,HG+J147 2组细胞活力、Bcl-2/Bax比值、p-AKT/AKT、p-ERK/ERK、p-CREB/CREB、BDNF蛋白表达升高(P<0.01),LDH活性、细胞凋亡率降低(P<0.01)。与HG+J147 2组比较,HG+J147 2+LY、HG+J147 2+U0组细胞活力、Bcl-2/Bax比值、p-AKT/AKT、BDNF蛋白表达降低(P<0.05或P<0.01),LDH活性、细胞凋亡率升高(P<0.05或P<0.01),HG+J1472+LY组p-ERK/ERK蛋白表达降低(P<0.05),HG+J1472+U0组p-CREB/CREB蛋白表达降低(P<0.05)。结论 J147能减轻HG诱导的SH-SY5Y细胞损伤,其机制可能与激活PI3K/AKT和ERK1/2信号通路,减少细胞凋亡相关。Objective To investigate the protective effect and mechanism of J147 on the injury of human neuroblastoma cells(SH-SY5Y) induced by high glucose(HG). Methods We established HG-induced SH-SY5Y cell injury model. Then SH-SY5Y cells were divided into blank control(Con) group HG group,HG+J147 0. 5 μmol/L(HG+J147 0. 5)group,HG+J147 1 μmol/L(HG+J147 1)group,HG+J147 2 μmol/L(HG+J147 2)group,HG+ PI3K/AKT inhibitor LY294002(LY) 10 μmol/L(HG+LY)group,HG+ERK1/2 inhibitor U0126(U0) 5 μmol/L(HG +U0)group,HG+J147 2 μmol/L+LY 10 μmol/L(HG+J147 2+LY)group,HG+J147 2 μmol/L+ U0 5 μmol/L(HG+ J147 2+U0)group. Cell viability was detected by MTS cell proliferation and toxicity detection kit;LDH activity was tested by lactate dehydrogenase kit;morphological changes of SH-SY5Y cells were evaluated by microscope;cell apoptosis was detected by flow cytometry;and apoptosis-related proteins(Bcl-2,Bax) and signaling pathway-related proteins(p-AKT,AKT,p-ERK1/2,ERK1/2,p-CREB,CREB,BDNF) were detected by Western blot. Results Compared with Con group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT,p-ERK/ERK,p-CREB/CREB and BDNF protein expressions decreased(P<0. 01),while LDH activity and apoptosis rate increased in HG group(P<0. 01). Compared with HG group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT,p-ERK/ERK,p-CREB/CREB and BDNF protein expressions increased(P<0. 01),while LDH activity and apoptosis rate decreased in HG+J147 2 group(P<0. 01). Compared with HG+J147 2 group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT and BDNF protein expression decreased(P<0. 05 or P<0. 01),while LDH activity and apoptosis rate increased(P<0. 05 or P<0. 01),the expression of p-ERK/ERK protein in HG+J147 2+LY group decreased(P<0. 05),and the expression of p-CREB/CREB protein in HG+J147 2+U0 group decreased in HG+J147 2+LY and HG+J147 2+U0groups(P<0. 05). Conclusions J147 can alleviate HG-induced SH-SY5Y cell damage,and the mechanism may be related to the activation of PI3K/AKT and ERK1/2 signaling and the reduction of apoptosis.

关 键 词：J147 SH-SY5Y细胞 高糖 细胞凋亡 

分 类 号：R587.2[医药卫生—内分泌] R745[医药卫生—内科学]