葛根芩连汤对2型糖尿病大鼠脂肪组织胆固醇及合成关键酶基因表达和甲基化水平影响的研究  

Effects of Gegen Qinlian Decoction(葛根芩连汤)on Cholesterol,Expressions and Methylation Levels of Genes Encoding Key Enzymes of Cholesterol Synthesis in Adipose Tissue of Type 2 Diabetic Rats

作　　者：李紫薇 刘欣宜 杨灵坤 张蒙雨 李昊天 陈嫦娥 章常华[1] 彭淑红[1,2] LI Ziwei;LIU Xinyi;YANG Lingkun;ZHANG Mengyu;LI Haotian;CHEN Chang'e;ZHANG Chanhua;PENG Shuhong(Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi,China;Jiangxi Province Key Laboratory of Traditional Chinese Medicine Etiopathogenisis,Nanchang 330004,Jiangxi,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]江西省中医病因生物学重点实验室,江西南昌330004

出　　处：《中华中医药学刊》2025年第2期173-181,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82060741);江西省主要学科学术和技术带头人培养项目(领军人才-学术类)(20232BCJ22022);江西中医药大学校级研究生创新专项(JZYC22S13);江西中医药大学中西医结合一级学科(江西省双一流学科)项目(zxyylxk20220103);江西中医药大学校级科技创新团队发展计划项目。

摘　　要：目的探讨葛根芩连汤(Gegen qinlian decoction,GQD)对2型糖尿病(Type 2 diabetes mellitus,T2DM)大鼠腹股沟脂肪组织(Inguinal white adipose tissue,iWAT)及附睾脂肪组织(Epididymal white adipose tissue,eWAT)胆固醇水平的影响,并从胆固醇合成关键酶的DNA甲基化角度探讨可能的机制。方法实验分为5组:对照组(CON)、2型糖尿病组(T2DM)、二甲双胍组(MET)、GQD低剂量组(GQDL)、GQD高剂量组(GQDH),灌胃给药14周。分光光度计法测定组织胆固醇水平,蛋白免疫印迹法(Western blotting,WB)、荧光定量PCR法(Quantitative Real-time PCR,QPCR)和亚硫酸盐测序PCR法(Bisulfite sequencing PCR,BSP)分别用于测定基因蛋白表达、mRNA表达和甲基化水平。结果MET和GQDH均能显著逆转T2DM中iWAT和eWAT胆固醇水平的降低;GQDL能有效逆转eWAT胆固醇水平的降低;GQDL和GQDH能够有效升高iWAT和eWAT中胆固醇合成关键酶3-羟基-3-甲基戊二酸单酰辅酶A还原酶(3-Hydroxy-3-methylglutaryl coenzyme A reductase,HMGCR)蛋白表达水平,但未有效调节角鲨烯环氧化酶(Squalene epoxidase,SQLE)蛋白表达水平;给药组均未有效升高胆固醇合成关键酶基因mRNA水平;Hmgcr基因在iWAT和eWAT中的CAs、eWAT中的CG3甲基化与血清空腹血糖(Fasting blood glucose,FBG)、糖化血清蛋白(Glycosylated serum protein,GSP)或胰岛素有相关性,而模型组eWAT的CG3甲基化显著降低,MET组eWAT的CAs甲基化显著升高。结论GQD能有效逆转T2DM大鼠脂肪组织胆固醇水平,可能不是通过调节胆固醇合成来实现。但GQD可能通过对HMGCR的转录后调控促进甲羟戊酸途径的非甾醇分支代谢。脂肪组织Hmgcr基因的CAs和CG3位点低甲基化可能与糖尿病发生密切相关。Objective To investigate the effects of Gegen Qinlian Decoction(葛根芩连汤,GQD)on cholesterol levels in inguinal adipose tissue(epididymal white adipose tissue,iWAT)and epididymal adipose tissue(epididymal white adipose tissue,eWAT)of type 2 diabetic rats and to explore the possible mechanism from the viewpoint of DNA methylation of genes encoding key enzymes of cholesterol synthesis.Methods Rats were divided into 5 groups:control group(CON),type 2 diabetes mellitus model group(T2DM),metformin group(MET),GQD low-dose and high-dose groups(GQDL,GQDH)which were given the corresponding medicines via gavage for 14 weeks.Choelsterol levels were determined by spectrophotometer.Western blot(WB),Quantitative Real-time PCR(QPCR)and Bisulfite sequencing PCR(BSP)were used to detect protein,mRNA expression and DNA methylation respectively.Results Both MET and GQDH could effectively reverse cholesterol levels of iWAT and eWAT in diabeteic rats.Moreover,GQDL could effectively reverse cholesterol levels of eWAT.GQDL and GQDH effectively elevated the protein expression levels of Hmgcr inWAT and eWAT,however,treatment groups didn't exhibit the increase of the mRNA expression of genes encoding key enzymes of cholesterol synthesis.CAs methylation of Hmgcr in iWAT and eWAT and CG3 methylation of Hmgcr in eWAT were correlated with FBG,GSP or insulin of serum.Furthermore,CG3 methylation of eWAT in T2DM group was significantly reduced,while CAs methylation of eWAT in MET group was significantly increased.Conclusion The effect of GQD or MET on reversing cholesterol level in adipose tissue of diabetic rats may not be achieved by regulating the biosynthesis of the cholesterol.However,GQD may promote the nonsterol branch of mevalonate pathway by post-transcriptional regulation of Hmgcr.The hypomethylation of CAs and CG3 in adipose tissue may be closely related to the progress of diabetes mellitus.

关 键 词：胆固醇合成 DNA甲基化 2型糖尿病 脂肪组织 

分 类 号：R289.5[医药卫生—方剂学]