目的 探讨补阳还五汤合参芪地黄汤化裁是否通过 m6A RNA 甲基化酶——甲基转移酶蛋白 3（METTL3） 调节 Snail 改善糖尿病肾病（DKD）小鼠肾组织损伤。方法 将 11～12 周龄 db/db 小鼠 24 只适应性喂养 1 周并 监测尿蛋白均阳性后随机分为模型组、中药组、西药组。db/m 组小鼠作为正常组，每组 8 只。中药组和西药 组分别予补阳还五汤合参芪地黄汤化裁和厄贝沙坦灌胃；正常组及模型组灌胃等体积蒸馏水，每日 1 次，连 续 12 周。之后处死小鼠，摘取肾脏，HE、Masson 染色观测肾组织病理改变。全自动生化分析仪检测小鼠血 糖（FBG）、总胆固醇（TC）、甘油三脂（TG）、尿素氮（BUN）、肌酐（Scr）、尿微量白蛋白（mALB）水平； qRT-PCR 和 Western Blot 法检测甲基转移酶蛋白 3（METTL3）、甲基转移酶蛋白 14（METTL14）、肾母细胞瘤 1 关联蛋白（WTAP）、ALKB 同源物 5（ALKBH5）、脂肪与肥胖相关蛋白（FTO）和 Snail 的蛋白及 mRNA 表达； Western Blot 法检测小鼠肾组织上皮-间充质转化（EMT）、纤连蛋白（FN）、波形蛋白（Vimentin）、α-平滑 肌肌动蛋白（α-SMA）表达；免疫荧光检测 E-cadherin、α-SMA 的表达；免疫共定位检测 METTL3 与 Snail 的 共表达。同时光镜观察肾脏病理形态学变化。结果 与正常组比较，模型组小鼠肾小球肥大，系膜基质增加， 基底膜增厚，囊腔变窄，间质炎性细胞浸润及纤维化（P＜0.05）；FBG、TC、TG、BUN、Scr 及 mALB 含量明 显升高（P＜0.01）；肾组织 METTL3、METTL14 和 WTAP 的蛋白和 mRNA 降低（P＜0.05），Snail mRNA 和 ALKBH5 的蛋白和 mRNA 升高（P＜0.05），而 FTO 的蛋白和 mRNA 无明显变化（P＞0.05）；同时 EMT 相关蛋 白 E-cadherin 降低（P＜0.05），Vimentin、α-SMA、FN 升高（P＜0.05）。与模型组比较，经补阳还五汤合参芪 地黄汤化裁中药干预后，糖尿病肾病小鼠肾脏病理损伤程度明显改善（P＜0.05）；TC、TG、BUN、Scr 及 mALB 明显降低（P＜0.01）；肾组织 m6A 水平恢复（P＜0.05），METTL3 的蛋白和 mRNA 升高（P＜0.05），Snail mRNA 水平降低（P＜0.01），同时抑制了肾组织 EMT 的发生（P＜0.05）；而 METTL14、ALKBH5、FTO、WTAP 的蛋白 和 mRNA 无明显变化（P＞0.05），对 FBG 水平也无明显影响（P＞0.05）。结论 补阳还五汤合参芪地黄汤化裁 可能通过 m6A RNA 甲基化酶 METTL3 调节 Snail 改善糖尿病肾病小鼠肾脏损伤，降低尿蛋白排泄，改善糖尿 病肾病肾间质纤维化的进展。

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.

R285.5

河北省自然科学基金资助项目（H2022423342）；河北省教育厅重点项目（ZD2021081）；河北省重点研发计划中医药创新专项（22377715OD）。