摘要
目的探讨雷公藤甲素(triptolide,TPL)对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾间质纤维化的作用及机制。方法随机选取6只雄性C57BL/6J小鼠作为Sham组,12只小鼠UUO建模成功后随机均分为模型组(UUO组)和TPL组。比较各组小鼠血肌酐(serum creatinine,SCr)、尿素氮(blood urea nitrogen,BUN)、体质量水平变化;术后14 d采集肾组织行HE和Masson染色观察各组肾脏病理损伤程度;免疫组化染色观察肾组织α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)及纤维连接蛋白(fibronectin,Fn)的表达。蛋白免疫印迹(Western blotting,Wb)方法检测NOD样受体热蛋白结构域蛋白3(nucleotide combined with structure of oligomerization domain receptor protein 3,NLRP3)、GSDMD、cGSDMD、IL-1β以及IL-18的蛋白表达。结果在1周时,UUO组和TPL组小鼠体质量与Sham组相比均明显下降,差异有统计学意义(P<0.05);在2周时,TPL组小鼠体质量与Sham组相比明显下降,差异有统计学意义(P<0.01),但与UUO组小鼠体质量之间无明显差异。BUN水平在3组间无差异;与Sham组比较,UUO组SCr水平明显升高[(15.680±1.508)μmol/L],差异具有统计学意义(P<0.01),给予TPL干预后,SCr水平明显下降[(12.550±3.004)μmol/L](P<0.05),差异具有统计学意义。HE染色结果显示,与Sham组比,UUO组小鼠肾小管明显扩张,小管萎缩,小管间质水肿,肾间质炎性细胞浸润增加,给予TPL后,肾组织病理损伤程度明显缓解。Masson染色结果显示,UUO组肾小管间质胶原沉积明显增加(36.350±5.183)%,给予TPL治疗后小管间质胶原沉积减少(20.820±3.290)%(P<0.01)。免疫组化染色结果显示,与Sham组比,UUO组小鼠α-SMA及Fn的表达升高,IOD值分别为(1.233±0.045)和(1.337±0.045);而给予TPL治疗后,其蛋白表达较UUO组下降,α-SMA的IOD值为(1.047±0.025),Fn的IOD值为(1.113±0.021)(P<0.05)。Wb结果显示,与Sham组比,UUO组小鼠NLRP3、cGSDMD、IL-1β以及IL-18的蛋白表达升高;而给予TPL治疗后,上述指标的蛋白表达较UUO组明显下降(P<0.05)。结论TPL可能通过抑制NLRP3介导的焦亡改善UUO小鼠肾间质纤维化。
Objective To investigate the effect of triptolide(TPL)on renal interstitial fibrosis in mice with unilateral ureteral obstruction(UUO)and its mechanism.Methods Six male C57BL/6J mice were randomly selected as the sham group,and 12 mice with UUO modeling were randomly divided into the model group(UUO)and the triptolide group(TPL).Changes in serum creatinine(SCr),blood urea nitrogen(BUN),and body weight were compared among the groups.Renal tissue specimens were collected at 14 d after UUO for HE and Masson staining.Immunohistochemistry staining was performed to observe the expression of α-smooth muscle actin(α-SMA)and fibronectin(Fn)in kidney tissues.Western blotting analysis was used to detect the protein expression levels of nucleotide combined with structure of oligomerization domain receptor protein 3(NLRP3),GSDMD,cGSDMD,IL-1β and IL-18.Results At week 1,the body weight of mice in the UUO and TPL groups significantly decreased compared with that in the sham group(P<0.05).Body weight reduced in the TPL group compared with that in the Sham group at week 2(P<0.01).There was no significant difference in body weight between the TPL and UUO groups.BUN levels did not differ significantly between the three groups.Compared with the sham group,the SCr level in the UUO group significantly increased[(15.680±1.508)μmol/L](P<0.01).A reduction in SCr level was observed following TPL administration[(12.550±3.004)μmol/L](P<0.05).HE staining showed that the renal tubules of mice in the UUO group were significantly dilated and atrophic,with interstitial edema and increased inflammatory cell infiltration,while the pathological damage of renal tissues was significantly alleviated after TPL treatment.Masson staining revealed that interstitial collagen deposition significantly increased in the UUO group(36.350±5.183)%(P<0.01)and reduced after TPL administration(20.820±3.290)%(P<0.01).Immunohistochemistry results demonstrated that the IOD levels ofα-SMA(1.233±0.045)and Fn(1.337±0.045)were higher in UUO group mice than in the sham group,while the IOD levels ofα-SMA(1.047±0.025)and Fn(1.113±0.021)were lower in the TPL group than in the UUO group(P<0.05).Western blotting analysis indicated that the expression levels of NLRP3,cGSDMD,IL-1βand IL-18 in the UUO group mice were higher than those in the sham group,while the protein expression levels of the above-mentioned indicators significantly decreased after TPL treatment(P<0.05).Conclusion TPL ameliorates renal interstitial fibrosis in mice with UUO by inhibiting NLRP3-mediated pyroptosis.
作者
魏琳婷
葛蓬勃
李艳
李柯
王引红
赵纬昊
崔晨凯
付荣国
WEI Linting;GE Pengbo;LI Yan;LI Ke;WANG Yinhong;ZHAO Weihao;CUI Chenkai;FU Rongguo(Department of Nephrology,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004;Department of General Surgery,The First Affiliated Hospital of Xi’an Medical College,Xi’an 710077,China)
出处
《西安交通大学学报(医学版)》
北大核心
2025年第6期947-952,共6页
Journal of Xi’an Jiaotong University(Medical Sciences)
基金
国家自然科学基金资助项目(No.82170697,No.82405169)
陕西省重点产业创新链(群)项目(No.2024SF-ZDCYL-03-05)
陕西省自然科学基金资助项目(No.2022JM-472)
中国医师协会医学循证专委会肾科学组PRO·润基金资助项目(No.KYJ202206-0003-3)。
