参芪复方对糖尿病GK大鼠肝脏组织mRNA、lncRNA表达谱的影响  

Effect of ShenqiCompound(参芪复方)on mRNA and IncRNA Expression Profiles in Liver of Diabetic GK Rats

作　　者：刘桠[1] 张翕宇 谢春光[1,2] 徐刚 彭思涵 LIU Ya;ZHANG Xiyu;XIE Chunguang;XU Gang;PENG Sihan(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,Sichuan,China;TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Chengdu 610072,Sichuan,China)

机构地区：[1]成都中医药大学附属医院,四川成都610072 [2]代谢性疾病中医药调控四川省重点实验室,四川成都610072

出　　处：《中华中医药学刊》2024年第1期46-51,I0008-I0012,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82104813);四川省博士后创新人才支持项目(BX202205);四川省科技创新苗子工程项目(2022036);成都中医药大学附属医院基金项目(22HL02)。

摘　　要：目的应用全转录组测序与实时聚合酶链锁反应(Real-time polymerase chain reaction,RT-qPCR)技术,探索参芪复方调控糖尿病GK大鼠肝脏信使RNA(messenger RNA,mRNA)、长链非编码RNA(long non-coding RNA,lncRNA)表达谱的机制研究。方法采用高脂高糖饲料建立2型糖尿病模型,将GK大鼠随机分为模型组、参芪复方组和西药组。另将10只Wistar大鼠设为空白组,予普通饲料喂养。参芪复方组大鼠予参芪复方浸膏灌胃,西药组予西格列汀混悬液灌胃,模型组及空白组灌服生理盐水。干预12周,每周检测空腹血糖(fasting blood glucose,FBG)。运用HE染色法观察大鼠肝脏组织病理形态,在空白组、模型组和参芪复方组中每组随机选取4只大鼠进行全转录组测序,构建mRNA、lncRNA差异表达谱,分析差异基因生物学功能,并进行RT-qPCR验证。结果与空白组相比,糖尿病GK大鼠FBG显著升高(P<0.05),模型组大鼠肝脏可见肝细胞排列紊乱,边界模糊,出现弥漫空泡状改变,呈中度脂肪变性,伴见炎性浸润、弥漫水肿;与模型组相比,参芪复方组大鼠FBG在12周时明显降低(P<0.05),肝脏组织排列较整齐,脂质沉积、细胞水肿及炎细胞浸润明显减轻。全转录组结果显示,与空白组相比,模型组大鼠mRNA、lncRNA表达谱存在显著差异,决定了糖尿病大鼠生理病理上的差异表现。参芪复方可广泛调控mRNA、lncRNA的差异表达,富集分析显示参芪复方通过调控多条内分泌系统及代谢过程相关信号通路改善糖尿病,包括胰岛素分泌、非酒精性脂肪性肝病、胆固醇代谢、鞘脂代谢、胰岛素抵抗等信号通路。RT-qPCR结果显示,基因Irf1、Trim30、Tmcc3、Insig1与转录组结果一致,转录组准确性较高。结论参芪复方发挥调节血糖及改善肝脏脂质沉积、水肿的作用,可能与广泛调控mRNA、lncRNA的差异表达有关。Objective To explore the mechanism of Shenqi Compound(参芪复方)regulating the expression profiles of mRNA and lncRNA in the liver of diabetic GK rats by whole transcriptome sequencing and RT-qPCR.Methods Type 2 diabetes model was prepared by high fat and high sugar diet,and the GK rats were randomly divided into model group,Shenqi Compound group and western medicine group,and fed with high-fat and high-sugar diet.In addition,another 10 Wistar rats were set as the blank group and fed with the ordinary diet.The rats in Shenqi Compound group were given Shenqi Compound extract by gavage,those in the western medicine group were given sitagliptin suspension by gavage,and those in model group and blank group were given normal saline by gavage.After 12 weeks of intervention,fasting blood glucose(FBG)was measured weekly.HE staining was used to observe the pathological morphology of liver tissue of the rats.Four rats were randomly selected from each group of the blank group,model group and Shenqi Compound group for whole transcriptome sequencing.The differential expression profiles of mRNA and lncRNA were constructed,and the biological functions of the differential genes were analyzed and verified by RT-qPCR.Results Compared with that of the blank group,the FBG level of diabetic GK rats was significantly increased(P<0.05).The liver of the model group showed disordered hepatocyte arrangement,blurred boundary,diffuse vacuolar changes,moderate steatosis,inflammatory infiltration and diffuse edema.Compared with that in the model group,the FBG level of rats in the Shenqi Compound group was significantly decreased at 12 weeks(P<0.05),the liver tissue was arranged neatly,and steatosis,cell edema and inflammatory cell infiltration were significantly reduced.The whole transcriptome results showed that compared with those of the blank group,the mRNA and lncRNA expression profiles of the model group were significantly different,which determined the physiological and pathological differences of the diabetic rats.Shenqi Compound ca

关 键 词：参芪复方 2型糖尿病 长链非编码RNA(lncRNA) 信使RNA(mRNA) 

分 类 号：R289.5[医药卫生—方剂学]