摘要
目的:探究微小RNA(miRNA)-370沉默对酒精性肝病(ALD)大鼠的作用及机制。方法:32只Sprague-Dawley(SD)大鼠随机分为4组:正常组(普通饲料)、模型组(40%酒精灌胃+隔日高脂饲料饲养)、空载组(40%酒精灌胃+隔日高脂饲料饲养+尾静脉注射含空载质粒的腺病毒)和沉默组(40%酒精灌胃+隔日高脂饲料饲养+尾静脉注射含si-miRNA-370质粒的腺病毒),每组各8只。用40%酒精灌胃联合隔日高脂饲料饲养法构建ALD大鼠模型。酶联免疫吸附法检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)表达水平;实时荧光定量PCR法检测miRNA-370、沉默信息调节因子6(SIRT6)mRNA和Nrf2 mRNA表达水平;蛋白质免疫印迹法检测SIRT6和Nrf2表达水平。结果:与正常组比较,模型组SOD、GSH-Px、SIRT6 mRNA、Nrf2 mRNA、SIRT6和Nrf2水平降低,丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、甘油三酯(TG)、MDA和miRNA-370水平升高,差异均有统计学意义(q=10.306、14.839、9.040、10.645、5.836、8.775、11.098、10.066、8.569、4.976、12.435,均P<0.05)。与空载组比较,沉默组SOD、GSH-Px、SIRT6 mRNA、Nrf2 mRNA、SIRT6和Nrf2水平升高,ALT、AST、TG和miRNA-370水平降低,差异均有统计学意义(q=5.731、9.537、14.524、29.569、8.888、16.233、8.144、6.818、5.329、21.317,均P<0.05)。结论:MiRNA-370沉默可改善ALD大鼠肝脏损伤,升高SOD和GSH-Px水平,其机制可能与调控SIRT6和Nrf2表达有关。
Objective:To explore the effect and mechanism of microRNA(miRNA)-370 silencing on rats with alcoholic liver disease(ALD).Methods:Thirty-two Sprague-Dawley(SD)rats were randomly divided into 4 groups:normal group(common feed),model group(40%alcohol gavage+high-fat feed feeding every other day),no-load group(40%alcohol gavage+high-fat feed feeding every other day+tail vein injection of adenovirus containing empty plasmid)and silent group(40%alcohol gavage+high-fat feed feeding every other day+tail vein injection of adenovirus containing si-miRNA-370 plasmid),8 rats in each group.An ALD rat model was constructed using 40%alcohol gavage combined with high-fat feed every other day.Enzyme-linked immunosorbent assay was used to detect the expression levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA).Real-time fluorescence quantitative PCR method was used to detect miRNA-370 and SIRT6 mRNA and Nrf2 mRNA expression levels.Western blotting was used to detect the expression levels of SIRT6 and Nrf2.Results:Compared with the normal group,the levels of SOD,GSH-Px,SIRT6 mRNA,Nrf2 mRNA,SIRT6 and Nrf2 in the model group were decreased,and the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG),MDA and miRNA-370 were increased,the differences were statistically significant(q=10.306,14.839,9.040,10.645,5.836,8.775,11.098,10.066,8.569,4.976,12.435,all P<0.05).Compared with the no-load group,the levels of SOD,GSH-Px,SIRT6 mRNA,Nrf2 mRNA,SIRT6,and Nrf2 in the silent group increased,while the levels of ALT,AST,TG and miRNA-370 decreased,the differences were statistically significant(q=5.731,9.537,14.524,29.569,8.888,16.233,8.144,6.818,5.329,21.317,all P<0.05).Conclusion:MiRNA-370 silencing can improve liver injury in rats with ALD,increase the levels of SOD and GSH-Px.The mechanism may be related to the regulation of SIRT6 and Nrf2 expression.
作者
辛小敏
王秀敏
黄宏春
XIN Xiao-min;WANG Xiu-min;HUANG Hong-chun(Department of Gastroenterology,Anyang People′s Hospital,Anyang455000,China)
出处
《天津医科大学学报》
2021年第2期108-111,共4页
Journal of Tianjin Medical University
