摘要
该研究探索了辛酸钠对骨骼肌细胞缺糖缺氧/再灌注(OGD/Rep)损伤的保护作用。利用锥虫蓝染色法测定6种不同浓度的辛酸钠培养液对正常培养骨骼肌细胞24 h存活率的影响。随后采用缺糖缺氧后复糖复氧的方法构建骨骼肌细胞OGD/Rep损伤模型,将细胞随机分为对照组、OGD/Rep组、0.25 mmol/L辛酸钠组和0.50 mmol/L辛酸钠组。CCK8法测定各组细胞活性,检测各组细胞中乳酸脱氢酶(LDH)、超氧化物(·O_(2)^(-))以及超氧化物歧化酶(SOD)水平;JC-1荧光探针测定各组细胞线粒体膜电位水平;TUNEL染色法检测各组细胞凋亡数目,Western blot测定各组细胞Bax、Bcl-2、Mfn-2、Drp-1蛋白表达水平。结果表明,6种浓度辛酸钠处理组中,0.25 mmol/L辛酸钠组、0.50 mmol/L辛酸钠组与对照组细胞存活率差异无统计学意义(P>0.05)。与对照组相比,OGD/Rep组细胞活性降低,细胞LDH水平升高,细胞中·O_(2)^(-)产生量增加,SOD活性降低,线粒体膜电位降低以及细胞凋亡数目增多(P<0.05)。与OGD/Rep组相比,0.25 mmol/L辛酸钠组、0.50 mmol/L辛酸钠组细胞活性均增强,细胞LDH水平下降(P<0.05);0.25 mmol/L辛酸钠组·O_(2)^(-)水平有下降趋势,但差异无统计学意义(P>0.05);0.25 mmol/L辛酸钠组SOD活性增强(P<0.05)。0.25 mmol/L和0.50 mmol/L辛酸钠组与OGD/Rep组相比线粒体膜电位增加,细胞凋亡数目减少。与对照组相比,OGD/Rep组Bcl-2/Bax蛋白值降低,Drp-1和Mfn-2蛋白表达量降低;而与OGD/Rep组相比,0.25 mmol/L、0.50 mmol/L辛酸钠组Bcl-2/Bax蛋白值升高,Drp-1和Mfn-2蛋白表达量增加。研究表明,早期应用辛酸钠可减轻OGD/Rep引起的骨骼肌细胞损伤,抑制细胞的过氧化状态并减少细胞凋亡,其机制可能与辛酸钠调节线粒体结构动态平衡相关。
This work was to investigate the protective effect of sodium octanoate on OGD/Rep(oxygen and glucose deprivation/reperfusion)injury of skeletal muscle cells.A trypan-blue assay was used to determine the effects of six different concentrations of sodium octanoate on the 24 h survival rate of skeletal muscle cells.The OGD/Rep model of skeletal muscle cells was constructed by the compound sugar and reoxygenation method after glucose and oxygen deprivation.The cells were randomly divided into the control group,the OGD/Rep group,the 0.25 mmol/L sodium octanoate group and the 0.50 mmol/L sodium octanoate group.Cell activity was determined by CCK8 assay,and the levels of LDH(lactate dehydrogenase),·O_(2)^(-)(superoxide)and SOD(superoxide dismutase)in each group were detected.The mitochondrial membrane potential was measured using the fluorescent probe JC-1.Apoptotic cells were detected by TUNEL assay.Western blot was used to detect the expression of Bax,Bcl-2,Mfn-2 and Drp-1 proteins.Among the groups treated with six concentrations of sodium octanoate,only the 0.25 mmol/L sodium octanoate group and 0.50 mmol/L sodium octanoate group had no statistically significant difference in cell survival rate with the control group(P>0.05).Compared with the control group,OGD/Rep group showed lower cell viability,higher LDH level,higher·O_(2)^(-)production,lower SOD activity,lower mitochondrial membrane potential and more apoptotic cells(P<0.05).Compared with the OGD/Rep group,the cell activity of the 0.25 mmol/L and 0.50 mmol/L sodium octanoate groups were increased,and the LDH release decreased(P<0.05);the·O_(2)^(-)of the 0.25 mmol/L sodium octanoate group showed a trend towards decrease but the difference was not significant statistically(P>0.05);SOD activity increased in 0.25 mmol/L sodium octanoate group(P<0.05).Compared with OGD/Rep group,the mitochondrial membrane potential increased and the number of apoptotic cells decreased in 0.25 mmol/L and 0.50 mmol/L sodium octanoate groups.Compared with the control group,the ratio of Bcl-2/Bax protein and the expression of Drp-1 and Mfn-2 proteins decreased in OGD/Rep group,while the ratio of Bcl-2/Bax protein and the expression of Drp-1 and Mfn-2 proteins increased in 0.25 mmol/L and 0.50 mmol/L sodium octanoate groups compared with OGD/Rep group.This study suggests that the early application of sodium octanoate can reduce the OGD/Rep-induced injury of skeletal muscle cells,reduce the peroxidation damage and reduce the apoptosis of skeletal muscle cells.The mechanism may be related to the regulation of sodium octanoate on the dynamic balance of mitochondrial structure.
作者
刘少云
蒋相康
徐杰丰
何露
张茂
LIU Shaoyun;JIANG Xiangkang;XU Jiefeng;HE Lu;ZHANG Mao(Key Laboratory of the Diagnosis and Treatment of Severe Trauma and Burn of Zhejiang Province,Zhejiang Province Clinical Research Center for Emergency and Critical Care Medicine,Department of Emergency Medicine,the Second Affiliated Hospital School of Medicine,Hangzhou 310009,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2021年第12期2353-2360,共8页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:82072126)资助的课题。
关键词
辛酸钠
骨骼肌细胞
缺糖缺氧/再灌注
sodium octanoate
skeletal muscle cells
oxygen and glucose deprivation/reperfusion
