左归降糖清肝方对糖尿病性脂肪肝小鼠PGC-1α和DNMT3A表达的影响  

Effects of Zuogui Jiangtang Qinggan Formula on PGC-1αand DNMT3A expressions of liver in diabetic fatty liver mice

作　　者：彭泉 钟雁城 杨霄旭 成细华[1] PENG Quan;ZHONG Yancheng;YANG Xiaoxu;CHENG Xihua(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;College of Acupuncture and Massage,Changchun University of Chinese Medicine,Changchun,Jilin 130500,China)

机构地区：[1]湖南中医药大学,湖南长沙4102081 [2]长春中医药大学针灸推拿学院,吉林长春130500

出　　处：《湖南中医药大学学报》2023年第12期2171-2176,共6页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金区域联合创新研究重点支持项目(U21A20411);湖南省自然科学基金项目(2021JJ30491);湖南省教育厅重点项目(20A376);湖南省卫健委重点项目(A202303066904);湖南省教育厅一般项目(19C1385);湖南中医药大学大学生创新创业训练计划(2021-128,2022-153);湖南中医药大学一流学科《基础医学》项目;湖南省国内一流培育学科中西医结合学科项目;中西医结合病原生物学湖南省重点学科。

摘　　要：目的探讨左归降糖清肝方对高脂饮食2型糖尿病转基因MKR鼠脂肪肝发生中过氧化物酶体增殖活化受体γ共激活因子-1α(peroxisome proliferation-activated receptor-γ-coactivator 1α,PGC-1α)和DNA甲基转移酶3A(DNA methyltransferase 3A,DNMT3A)表达的影响。方法24只MKR鼠随机分为对照组、高脂模型组和左归降糖清肝方(中药方)组;高脂模型组和中药方组均以高脂饲料喂养8周,对照组以基础饲料喂养;中药方组高脂饲料喂养4周后,以中药汤剂灌胃干预治疗4周,其他组灌胃等量的蒸馏水。采用电化学法检测空腹血糖;试剂盒测定肝组织甘油三酯(triglyceride,TG)和游离脂肪酸(free fatty acid,FFA)含量;HE染色观察肝组织形态结构及病理变化;qRT-PCR检测小鼠肝组织PGC-1α、线粒体转录因子A(mitochondrial transcriptionfactor A,TFAM)、核呼吸因子-1(nuclear respiratory factor 1,NRF1)和DNMT3A mRNA表达;Western blot和免疫组织化学检测PGC-1α和DNMT3A蛋白表达。结果高脂饮食诱发MKR鼠形成糖尿病性脂肪肝,肝细胞有大小不一的脂滴,呈脂肪性变;与对照组比较,高脂模型组小鼠血糖、肝指数和肝组织TG和FFA含量显著上升(P<0.01),肝脏PGC-1α、NRF1和TFAM的mRNA及PGC-1α蛋白表达显著下降(P<0.01),而DNMT3A mRNA和蛋白表达均显著上升(P<0.01)。与高脂模型组比较,左归降糖清肝方降低了高脂饮食MKR鼠血糖、肝指数和肝组织TG和FFA含量(P<0.01),上调了肝脏PGC-1α、NRF1和TFAM的mRNA表达,以及PGC-1α蛋白表达(P<0.01),下调了DNMT3A的mRNA和蛋白表达(P<0.01)。结论左归降糖清肝方通过调节肝脏PGC-1α及其通路相关基因表达,缓解糖尿病性脂肪肝小鼠肝脏脂肪性变,其机制可能通过抑制DNA甲基转移酶3A表达,减少PGC-1αDNA的甲基化相关。Objective To investigate the effects of Zuogui Jiangtang Qinggan Formula(ZGJTQGF)on the expression of peroxisome proliferation-activated receptor-γ-coactivator 1α(PGC-1α)and DNA methyltransferase 3A(DNMT3A)in fatty liver development of transgenic MKR mice with type 2 diabetes mellitus following a high-fat diet.Methods A total of 24 MKR mice were randomly divided into control group,high fat model group and ZGJTQGF group.High fat model group and ZGJTQGF group were fed with high fat diet for eight weeks,and control group was fed with basic diet.After feeding high-fat diet for four weeks,the Chinese medicine group was treated with Chinese medicine decoction for four weeks,and the other groups were given the same amount of distilled water.Fasting blood glucose was determined by electrochemical method.The content of triglyceride(TG)and free fatty acid(FFA)in liver tissue was determined by the kit.HE staining was used to observe the morphological structure and pathological changes of liver tissue.The mRNA expressions of PGC-1α,mitochondrial transcription factor A(TFAM),nuclear respiratory factor-1(NRF1),and DNMT3A in mice liver were tested by qRT-PCR,and the protein expressions of PGC-1αand DNMT3A were checked by Western blot and immunohistochemical analysis.Results High-fat diet induced the formation of nonalcoholic fatty liver in MKR mice.The liver cells had fat droplets of different sizes,which showed fatty changes.Compared with the control group,the blood glucose,liver index,TG and FFA content of liver tissue in the high-fat model group significantly increased(P<0.01),the PGC-1α,NRF1,and protein expression of mRNA and PGC-1αsignificantly decreased(P<0.01;the mRNA and protein expressions of DNMT3A significantly increased(P<0.01).Compared with high-fat model group,ZGJTQGF group had lower blood glucose,liver index,and TG and FFA content in liver tissues of high-fat diet MKR mice(P<0.01),up-regulated mRNA expressions of PGC-1α,NRF1,and TFAM in liver and the PGC-1αprotein expression(P<0.01),down-regulated mRNA a

关 键 词：左归降糖清肝方 糖尿病性脂肪肝 PGC-1Α DNMT3A 甲基化 

分 类 号：R285.5[医药卫生—中药学]