摘要
目的 观察萆薢分清颗粒对单侧输尿管梗阻大鼠肾纤维化相关蛋白雷帕霉素靶蛋白(mTOR)、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(Col-Ⅰ)的影响,探讨萆薢分清颗粒的肾脏保护作用.方法 取60 只SPF级雄性SD大鼠,从中随机取10 只作为空白组,其余50 只大鼠采用单侧输尿管梗阻术构建肾纤维化模型.将造模成功大鼠随机分为模型组、氯沙坦组及萆薢分清高、中、低剂量组,每组10只,萆薢分清高、中、低剂量组分别给予13.12 mL/(kg·d)、6.56 mL/(kg·d)、3.28 mL/(kg·d)萆薢分清颗粒溶液灌胃,氯沙坦组给予氯沙坦2.62 mL/(kg·d)灌胃,空白组和模型组给予等量生理盐水灌胃,均1 次/d,连续灌胃4 周.末次灌胃结束后检测尿白蛋白与尿肌酐的比值(ACR)、24 h尿蛋白量及血肌酐(SCr)、血尿素氮(BUN)水平,HE染色、Masson染色观察肾脏病理形态,免疫组化法及Western blot法分别检测大鼠肾脏组织中mTOR、α-SMA、Col-Ⅰ表达情况.结果 与空白组比较,模型组大鼠ACR、24 h尿蛋白量及SCr、BUN水平均明显升高(P均＜0.05);肾脏发生明显纤维化病变;肾组织中mTOR、α-SMA、Col-Ⅰ阳性表达平均光密度值和蛋白相对表达量均明显升高(P均＜0.05).与模型组比较,氯沙坦组与萆薢分清高、中剂量组大鼠ACR、24 h尿蛋白量及SCr、BUN 水平均明显降低(P 均＜0.05);肾纤维化病变较轻;肾组织中mTOR、α-SMA、Col-Ⅰ阳性表达平均光密度值和蛋白相对表达量均明显降低(P均＜0.05).结论 萆薢分清颗粒可减轻单侧输尿管梗阻大鼠肾纤维化,保护残存肾功能,机制可能与下调mTOR、α-SMA、Col-Ⅰ表达有关.
Abstract
Objective It is to observe the effect of Bixie Fenqing granules on renal fibrosis related proteins mammalian target of rapamycin(mTOR),α-smooth muscle actin(α-SMA)and collagen type Ⅰ(Col-Ⅰ)in unilateral ureteral ob-struction rats,and to explore the protective effect of this medicine on renal fibrosis.Methods Sixty SPF male SD rats were selected,in which 10 rats were randomly selected as blank group,and the remaining 50 rats were treated with unilateral u-reteral obstruction to establish models of renal fibrosis.The successfully modeled rats were randomly divided into model group,losartan group,and high dose,medium dose and low dose groups of Bixie Fenqing granule,with 10 rats in each group.The high dose,medium dose and low dose groups of Bixie Fenqing granule were respectively given Bixie Fenqing granule 13.12 mL/(kg·d),6.56 mL/(kg·d),3.28 mL/(kg·d)by gavage,the losartan group was given losartan 2.62 mL/(kg·d)by gavage,and the blank group and model group were given the same volume of normal saline by ga-vage,all once daily,continuously treated for 4 weeks.At the end of the last gavage,the albumin/urine creatinine ratio(ACR),24-hour urinary protein quantification(24 h-UPQ)and the levels of serum creatinine(SCr)and blood urea nitro-gen(BUN)were detected,the pathological morphology of the kidneys was observed by HE staining and Masson staining,and the expressions of mTOR,α-SMA,and Col-Ⅰ in renal tissues of the rats were detected by immunohistochemistry and Western blot methods.Results Compared with the blank group,the ACR,24 h-UPQ,SCr and BUN of rats in the model group were significantly increased(all P＜0.05),the renal fibrosis was obvious,and the mean optical density values and relative protein expression of mTOR,α-SMA,and Col-Ⅰ in renal tissues were significantly increased(all P＜0.05).Compared with the model group,the ACR,24 h-UPQ,SCr and BUN of rats in the losartan group,and high dose,medium dose groups of Bixie Fenqing granule were significantly decreased(all P＜0.05),the renal fibrosis was improved,and the mean optical density values and relative protein expression of mTOR,α-SMA,and Col-Ⅰin renal tissues were significantly decreased(all P＜0.05).Conclusion Bixie Fenqing granules can attenuate renal fibrosis and protect residual renal func-tion in rats with unilateral ureteral obstruction,and the mechanism may be related to down-regulating the expressions of mTOR,α-SMA,and Col-Ⅰ.
基金项目
甘肃省自然科学基金项目(21JR7RA577)
甘肃省名中医传承工作室建设项目(马鸿斌)(甘财社[2022]105号)
维普
万方数据