中药新药与临床药理2025，Vol.36Issue(5)：718-728,11.DOI:10.19378/j.issn.1003-9783.2025.05.007
补阳还五汤合参芪地黄汤化裁通过m6A RNA甲基化酶METTL3调节Snail改善小鼠糖尿病肾病的肾脏损伤
Modified Buyang Huanwu Decoction Combined with Shenqi Dihuang Decoction Ameliorates Renal Injury in Diabetic Kidney Disease Mice by Regulating Snail via m6A RNA Methyltransferase METTL3
摘要
Abstract
Objective To investigate whether the modified Buyang Huanwu Decoction combined with Shenqi Dihuang Decoction ameliorates renal injury in diabetic kidney disease(DKD)mice by regulating Snail through the m6A RNA methyltransferase METTL3.Methods Twenty-four db/db mice aged 11-12 weeks were adaptively fed for 1 week,and those with positive urinary protein were randomly divided into model,traditional Chinese medicine(TCM),and western medicine groups.Db/m mice served as the normal group,with 8 mice in each group.The TCM group and western medicine group were administered the modified Buyang Huanwu Decoction combined with Shenqi Dihuang Decoction and Irbesartan,respectively,by gavage.The normal and model groups received an equal volume of distilled water by gavage once daily for 12 weeks.Afterward,the mice were euthanized,and kidneys were harvested.Renal histopathological changes were observed using HE and Masson staining.Fasting blood glucose(FBG),total cholesterol(TC),triglycerides(TG),blood urea nitrogen(BUN),serum creatinine(Scr),and urinary microalbumin(mALB)levels were measured using an automatic biochemical analyzer.The protein and mRNA expression of METTL3,METTL14,WTAP,ALKBH5,FTO,and Snail was detected by qRT-PCR and Western Blot.Western Blot was also used to assess the expressiond of epithelial-mesenchymal transition(EMT)-related proteins,including fibronectin(FN),Vimentin,and α-smooth muscle actin(α-SMA).Immunofluorescence was employed to detect E-cadherin and α-SMA expression,while co-immunolocalization was used to examine the co-expression of METTL3 and Snail.Renal histomorphological changes were observed under light microscopy.Results Compared with the normal group,the model group exhibited glomerular hypertrophy,increased mesangial matrix,thickened basement membrane,narrowed capsular space,interstitial inflammatory cell infiltration,and fibrosis(P＜0.05).FBG,TC,TG,BUN,Scr,and mALB levels were significantly elevated(P＜0.01).The protein and mRNA expression of METTL3,METTL14,and WTAP in renal tissues was decreased(P＜0.05),while Snail mRNA and ALKBH5 mRNA and protein expression was increased(P＜0.05).FTO expression showed no significant change(P＞0.05).EMT-related protein E-cadherin was decreased(P＜0.05),while Vimentin,α-SMA,and FN were increased(P＜0.05).Compared with the model group,the TCM intervention significantly ameliorated renal pathological damage in DKD mice(P＜0.05).TC,TG,BUN,Scr,and mALB levels were significantly reduced(P＜0.01).Renal m6A levels were restored(P＜0.05),protein and mRNA expression of METTL3 was increased(P＜0.05),and protein and mRNA level of Snail was decreased(P＜0.01).EMT in renal tissues was also inhibited(P＜0.05).However,the protein and mRNA expression of METTL14,ALKBH5,FTO,and WTAP showed no significant changes(P＞0.05),and FBG levels were unaffected(P＞0.05).Conclusion The modified Buyang Huanwu Decoction combined with Shenqi Dihuang Decoction may ameliorate renal injury in DKD mice by regulating Snail through the m6A RNA methyltransferase METTL3,reducing urinary protein excretion,and mitigating the progression of renal interstitial fibrosis in diabetic kidney disease.关键词
糖尿病肾脏疾病/补阳还五汤合参芪地黄汤化裁/RNA甲基化/上皮-间充质转化/肾间质纤维化/甲基转移酶蛋白3/Snail/小鼠Key words
diabetic kidney disease/modified Buyang Huanwu Decoction combined with Shenqi Dihuang Decoction/RNA methylation/epithelial-mesenchymal transition/renal interstitial fibrosis/methyltransferase-like protein 3/Snail/mice分类
医药卫生引用本文复制引用
刘妍,杨帆,黄家安,张晓云,王月华..补阳还五汤合参芪地黄汤化裁通过m6A RNA甲基化酶METTL3调节Snail改善小鼠糖尿病肾病的肾脏损伤[J].中药新药与临床药理,2025,36(5):718-728,11.基金项目
河北省自然科学基金资助项目(H2022423342) （H2022423342）
河北省教育厅重点项目(ZD2021081) （ZD2021081）
河北省重点研发计划中医药创新专项(22377715OD). （22377715OD）
