摘要
目的探讨四味地黄醇提取物对高糖作用的心肌细胞凋亡的影响及作用机制。方法实验设置对照组、模型组、模型+四味地黄醇提取物-L组、模型+四味地黄醇提取物-M组、模型+四味地黄醇提取物-H组、模型+miR-NC组、模型+miR-26b组、模型+四味地黄醇提取物-M+anti-miR-NC组、模型+四味地黄醇提取物-M+anti-miR-26b组。流式细胞术检测细胞凋亡;乳酸脱氢酶(lactate dehydrogenase,LDH)试剂盒、超氧化物歧化酶(superoxide dismutase,SOD)试剂盒、丙二醛(malondialdehyde,MDA)试剂盒分别检测LDH、SOD活性及MDA含量;实时荧光定量PCR(RT-qPCR)检测miR-26b和MAPK mRNA表达水平;荧光素酶报告实验检测miR-26b和MAPK的靶向关系。结果与对照组相比,模型组心肌细胞凋亡率显著升高,MDA含量、LDH活性显著升高,SOD活性显著降低,miR-26b表达水平显著降低,MAPK mRNA表达水平显著升高(P<0.05)。低、中、高浓度四味地黄醇提取物处理可降低细胞凋亡率和MDA含量、LDH活性,提高SOD活性,提高miR-26b表达水平,降低MAPK mRNA表达水平(P<0.05)。过表达miR-26b抑制高糖诱导的心肌细胞凋亡和氧化应激的产生。抑制miR-26b逆转了四味地黄醇提取物对高糖作用的细胞H9C2凋亡和氧化应激的抑制作用。miR-26b靶向调控MAPK。结论四味地黄醇提取物可抑制高糖作用的细胞H9C2凋亡和氧化应激,其机制可能与miR-26b及MAPK表达有关。
Objective To investigate the effect of ethanol extracts of Siwei Dihuang on the apoptosis of cardiomyocytes and its mechanism. Methods The experiment set the control group,model group,model group + ethanol extracts of Siwei Dihuang-L group, model group + ethanol extracts of Siwei Dihuang-M group, model group + ethanol extracts of Siwei Dihuang-H group,model group + miR-NC group,model group + miR-26b group model group + ethanol extracts of Siwei Dihuang-M + anti-miR-NC group, and model group + ethanol extracts of Siwei Dihuang-M + anti-miR-26b group. Flow cytometry was used to detect apoptosis;lactate dehydrogenase(LDH)kit,superoxide dismutase(SOD)kit,and malondialdehyde(MDA)kit were used to detect LDH,SOD activity and MDA content,respectively. Real-time quantitative PCR(RT-qPCR)was used to detect miR-26b and MAPK mRNA levels;luciferase report experiment was conducted to detect the targeting relationship between miR-26b and MAPK. Results Compared with those in control group, the apoptosis rate of the cardiomyocytes of the model group was significantly increased, MDA content and LDH activity were significantly increased, the activity of SOD was significantly decreased, miR-26b expression was significantly decreased, but MAPK mRNA expression was significantly increased(P<0. 05). Treatment with low,medium and high concentrations of ethanol extracts of Siwei Dihuang could reduce apoptosis rate,MDA content and LDH activity, increase SOD activity and increase miR-26b expression, and decrease MAPK mRNA expression(P<0. 05).Overexpression of miR-26b inhibited high glucose-induced cardiomyocyte apoptosis and oxidative stress. Inhibition of miR-26b reversed the inhibitory effect of ethanol extracts of Siwei Dihuang on H9C2 apoptosis and oxidative stress in high glucose-induced cells. miR-26b targeted and regulated MAPK. Conclusion Ethanol extracts of Siwei Dihuang can inhibit the apoptosis and oxidative stress of H9C2 cells in high glucose, and its mechanism may be related to the upregulation of miR-26b and MAPK expressions.
作者
刘彬
郭春棉
廉坤
屈曚
王小梅
韦程程
LIU Bin;GUO Chunmian;LIAN Kun;QU Meng;WANG Xiaomei;WEI Chengcheng(Department of Cardiovascular Medicine,The First Affiliated Hospital of Air Force Medical University,Xi’an 710032,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2021年第6期941-946,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
白求恩-默克糖尿病研究基金资助(G201744)
苏州工业园区心馨心血管健康基金会“进阶基金”资助(2019-CCAACCESS065)。
