三七颗粒调控miR-23b/ASK1/p38MAPK信号通路抑制NLRP3炎症小体合成改善肾纤维化的机制研究  

Study on the Mechanism of Sanqi Granules Regulating the miR-23b/ASK1/p38MAPK Signal Pathway to Inhibit NLRP3 Inflammatory Body Synthesis and Antagonise Renal Fibrosis

作　　者：黄雪霞[1] 吴金玉[1] 莫超 王小连 陆良喜 彭泓杰 李永聪 肖国居 HUANG Xuexia;WU Jinyu;MO Chao(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023)

机构地区：[1]广西中医药大学第一附属医院,南宁530023 [2]广西中医药大学,南宁530023

出　　处：《中国中西医结合肾病杂志》2025年第3期201-207,I0002,共8页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：广西自然科学基金面上项目(No.2020GXNSFAA297246);广西自然基金青年科学基金项目(No.2023GXNSFBA026185)。

摘　　要：目的:研究三七颗粒调控miR-23b/ASK1/p38MAPK信号通路抑制NLRP3合成改善肾纤维化的机制。方法:按随机数字表法将36只SD大鼠随机分为6组,即:正常对照组(A组)、模型组(B组)、miR-23b阴性腺病毒组(C组)、miR-23b过表达腺病毒组(D组)、三七颗粒组(E组)、厄贝沙坦组(F组),每组各6只。A组不造模,其余5组均采用尾静脉注射顺铂(3 mg/kg)诱导肾纤维化大鼠模型,C组造模成功后尾静脉注射miR-23b阴性腺病毒,D组造模成功后尾静脉注射miR-23b过表达腺病毒,其余4组尾静脉注射等量生理盐水。E组给予三七颗粒500 mg/kg灌胃,F组给予厄贝沙坦分散片150 mg/kg灌胃,其余4组均予等量生理盐水灌胃,每日给药1次,连续给药12周。第12周末处死各组大鼠,采集血液和肾组织。HE染色观察肾组织病理变化,全自动生化分析仪测定肾功能(Scr、BUN)水平,ELISA法检测血清炎症因子IL-1β、IL-18水平,Western blot检测肾组织miR23/ASK1/p38 MAPK信号通路关键因子(ASK1、p-p38MAPK)、NLRP3炎症小体(NLRP3、ASC、Caspase-1)、肾纤维化相关蛋白(FN、CollagenⅣ)表达的水平的变化,RT-qPCR检测肾组织中miR-23b的表达量。结果:与A组比较,B组大鼠肾间质大量炎性细胞浸润及间质纤维化,Scr、BUN水平显著升高(P<0.01),miR-23的表达明显降低(P<0.05),FN、CollagenⅣ、p-p38MAPK、NLRP3、ASC、Caspase-1蛋白的表达显著上升(P<0.01),IL-1β、IL-18水平显著升高(P<0.01)。与B组比较,E组大鼠肾间质炎性细胞和间质纤维化显著减少,肾功能(Scr、BUN)显著降低(P<0.01),miR-23b的表达明显升高(P<0.05),FN、CollagenⅣ、ASK1、p-p38MAPK、NLRP3、ASC、Caspase-1蛋白的表达均显著下调(P<0.01),IL-1β、IL-18水平显著降低(P<0.01),E组的疗效较D组的效果佳,与F组的疗效相当。结论:中药三七颗粒可能通过增加miR-23b过表达,阻断ASK1/p38MAPK信号通路的激活,抑制NLRP3炎症小体释放炎症因子,最终改善肾间质纤维化。Objective:To investigate the mechanism of Sanqi granules regulating the miR-23b/ASK1/p38MAPK signaling pathway,inhibiting NLRP3 synthesis,and antagonizing renal fibrosis.Methods:36 SD rats were randomly divided into 6 groups using a random number table method.Group A:Normal control group;Group B:Model group;Group C:miR-23b negative adenovirus group;Group D:miR-23b overexpression adenovirus group;Group E:Group 37;Group F:Irbesartan group,with 6 rats in each group.Group A did not create a model,while the other 5 groups were induced by tail vein injection of cisplatin(3 mg/kg)to induce renal fibrosis in rats.Group C received tail vein injection of miR-23b negative adenovirus after successful modeling,Group D received tail vein injection of miR-23b overexpressing adenovirus after successful modeling,and the remaining 4 groups received tail vein injection of an equal amount of physiological saline.Group E was given 500 mg/kg Sanqi granules by gavage,Group F was given 150 mg/kg Erbesartan dispersible tablets by gavage,and the other four groups were given equal amounts of physiological saline by gavage once a day for 12 consecutive weeks.On the 12th weekend,all groups of rats were euthanized and blood and kidney tissue were collected.HE staining and Masson staining were used to observe pathological changes in renal tissue,and the levels of Scr and BUN in renal function were measured using a fully automated biochemical analyzer,and ELISA was used to detect serum inflammatory factor IL-1β、IL-18.Western blot was used to detect the protein expression levels of miR-23/ASK1/p38MAPK signaling pathway related indicators(ASK1,p-p38MAPK),NLRP3 inflammasomes(NLRP3,ASC,Caspase-1),and renal fibrosis indicators(FN,CollagenⅣ)in renal tissue.RT qPCR was used to detect the expression level of miR-23b mRNA in renal tissue.Results:Compared with Group A,Group B rats had a large amount of inflammatory cell infiltration and interstitial fibrosis in the renal interstitium.The levels of Scr and BUN were significantly increased(P<0.01),whi

关 键 词：肾纤维化 三七颗粒 miR-23b/ASK1/p38MAPK信号通路 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]