摘要
目的 研究余甘子提取物对高糖诱导的PC12细胞损伤的保护作用和作用机制.方法 用高糖诱导PC12细胞损伤,并用余甘子提取物处理PC12细胞或通过细胞转染使miR-34a过表达.采用四甲基偶氮唑盐(MTT)检测细胞存活率,采用流式细胞术检测细胞凋亡率,采用蛋白质免疫印迹(Western blot)检测B细胞淋巴瘤-2相关X蛋白(Bax)和B细胞淋巴瘤-2(Bcl-2)蛋白表达.采用荧光法检测活性氧(ROS)水平.采用试剂盒检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)水平.采用实时荧光定量PCR(qRT-PCR)检测miR-34a的表达.结果 余甘子提取物明显提高高糖诱导的PC12细胞存活率、SOD和GSH-Px水平(LSD-t 值分别为-9.62、-9.64 和-12.04,P＜0.05),明显降低凋亡率,Bax/Bcl-2 比值,ROS、IL-6、IL-1β 和 miR-34a 水平(LSD-t 值分别为 24.64、25.63、20.63、11.21、13.47 和 13.88,P＜0.05).miR-34a 过表达明显降低高糖诱导的 PC12 细胞存活率、SOD和GSH-Px水平(LSD-t值分别为5.95、6.87和7.21,P＜0.05),明显提高凋亡率;Bax/Bcl-2比值,ROS、IL-6和IL-1β水平(LSD-t值分别为-9.64、-12.47、-6.69、-8.21和-8.99,P＜0.05).甘子提取物可逆转miR-34a过表达对高糖诱导的PC12细胞的影响.结论 余甘子提取物对高糖诱导的PC12细胞损伤具有保护作用,其机制可能与余甘子提取物抑制miR-34a表达进而降低氧化应激与炎症反应有关.
Abstract
Objective To study the protective effect of extracts of Phyllanthus emblica L.on damage of PC 12 cells induced by high glucose and its related mechanisms.Methods PC 12 cells were damaged by high glucose induction,and then treated with extracts of Phyllanthus emblica L.or miR-34a was overexpressed by cell transfection.Methyl thiazolyl tetrazolium assay(MTT)was used to detect cell survival rate.Flow cytometry was used to detect cell apoptosis rate.Western blot was used to detect the protein expression levels of B-lymphocytoma-2-associated X protein(Bax)and B-lymphocytoma-2(Bcl-2).Fluorescence method was used to detect reactive oxygen species(ROS)level.Relevant kits were used to detect the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),interleukin-6(IL-6)and interleukin-1β(IL-1β).Real-time quantitative PCR(qRT-PCR)was used to detect the expression levels of miR-34a.Results The survival rate,SOD and GSH-Px levels of PC 12 cells induced by high glucose were significantly increased by extracts of Phyllanthus emblica L.(LSD-t=-9.62,-9.64,-12.04,respectively,P＜0.05),while the apoptosis rate,Bax/Bcl-2 ratio,ROS,IL-6,IL-1β and miR-34a levels were significantly decreased(LSD-t=24.64,25.63,20.63,11.21,13.47,13.88,respectively,P＜0.05).Overexpression of miR-34a significantly reduced the survival rate,SOD and GSH-Px levels of PC 12 cells induced by high glucose(LSD-t=5.95,6.87,7.21,respectively,P＜0.05).While significantly increased the apoptosis rate,Bax/Bcl-2,ROS,IL-6 and IL-1βlevels(LSD-t=-9.64,-12.47,-6.69,-8.21,-8.99,respectively,P＜0.05).The extracts of Phyllanthus emblica L.could reverse the effect of miR-34a overexpression on high glucose-induced PC 12 cells.Conclusion The extracts of Phyllanthus emblica L.showed protective effect on damage of PC 12 cells induced by high glucose,and the mechanism may be related to the inhibition of miR-34a expression by extracts of Phyllanthus emblica L.and the reduction of oxidative stress and inflammation.
基金项目
江苏省人民医院浦口分院科技发展基金(2022)(KJ2022-5)
NETL
NSTL
万方数据