清热消瘕方延缓糖尿病肾脏病大鼠肾损伤作用机制

Potential mechanism of decoction for clearing heat and resolving mass in delaying kidney injury in diabetic kidney disease rats

目的探讨“肾络癥瘕”病机指导下的清热消瘕方对糖尿病肾脏病(DKD)大鼠肾损伤的保护作用及相关潜在机制。方法将24只雄性SD大鼠随机分为假手术组、模型组、清热消瘕方组和缬沙坦组,每组6只。除假手术组外,其余组大鼠以单侧肾切除联合链脲佐菌素二联法构建DKD模型。造模成功后,清热消瘕方组给予清热消瘕方颗粒剂10.2 g/(kg·d)灌胃,缬沙坦组给予缬沙坦8 mg/(kg·d)灌胃,假手术组和模型组给予等量蒸馏水灌胃,均连续灌胃16周。比较各组大鼠一般情况、体重、肾重体重比、肾功能相关指标、血糖及血脂水平,HE、PAS、Masson染色观察各组大鼠肾脏病理形态,免疫组化染色观察各组大鼠肾脏组织中谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(xCT)表达情况。结果与假手术组比较,模型组大鼠体重和肾脏组织中GPX4、xCT阳性表达面积百分比均明显降低(P均<0.05),肾重体重比、24 h尿蛋白总量、血尿素氮、血糖、血清胆固醇和三酰甘油水平均明显升高(P均<0.05);肾脏肾小球体积增大,肾小管上皮细胞广泛空泡变性坏死,肾小球硬化、纤维化改变明显。与模型组比较,清热消瘕方组大鼠肾脏组织中GPX4、xCT阳性表达面积百分比均明显升高(P均<0.05),肾重体重比、24 h尿蛋白总量、血尿素氮、血清胆固醇水平均明显降低(P均<0.05);肾小球系膜基质增生减少,纤维化程度有所缓解。结论清热消瘕方可有效减轻DKD肾损伤和上调肾脏GPX4、xCT表达,该方可能通过调控铁死亡延缓DKD的进展。

Objective It is to explore the protecting effect and the relevant potential mechanisms of decoction for clearing heat and resolving mass(DCHRM)under the guide of’pathogenesis of kidney collateral and abdominal mass’on renal injury in diabetic kidney disease(DKD)rats.Methods Twenty-four male SD rats were randomly divided into sham operation group,model group,DCHRM group and valsartan group,with 6 rats in each group.Except for the sham operation group,the rats in the other groups were given unilateral nephrectomy combined with streptozotocin by intraperitoneal injection to establish DKD models.After successful modeling,the DCHRM group was given DCHRM granules 10.2 g/(kg·d)by gavage,the valsartan group was given valsartan 8 mg/(kg·d)by gavage,and the sham operation group and model group were given an equal amount of distilled water by gavage,all continuously treated for 16 weeks.The general condition,body weight,kidney weight-to-weight ratio,renal function-related indexes,blood glucose and blood fat indexes of the rats in each group were compared,and the pathological morphology of the kidneys of the rats in each group were observed by HE,PAS,and Masson staining,and the expression of glutathione peroxidase 4(GPX4),solution carrier family 7 member 11(xCT)in the renal tissues of the rats in each group were observed by immunohistochemical staining.Results Compared with the sham operation group,the body weight and the percentage of positively expressed area of GPX4 and xCT in the renal tissues of the rats in the model group were significantly decreased(all P<0.05),and the kidney weight-to-weight ratio,the levels of total 24-h urinary protein,blood urea nitrogen,blood glucose,serum cholesterol and triacylglycerol were all significantly increased(all P<0.05),with increased volume of renal glomerular,obvious extensive vacuolar degeneration and necrosis of renal tubular epithelial cells,obvious glomerulosclerosis and fibrosis changes.Compared with the model group,the percentage of positively expressed area of GPX4 and xCT in renal tissues of the rats in the DCHRM group were significantly increased(P<0.05),and the renal weight-to-weight ratio,total 24-h urinary protein,blood urea nitrogen,and serum cholesterol level were all significantly decreased(all P<0.05),the proliferation of glomerular mesangial stroma was reduced,and the fibrosis was alleviated.Conclusion DCHRM could effectively alleviate renal injury in DKD,and it could up-regulate the expression of GPX4 and xCT,it suggesting that DCHRM might delay the progression of DKD by regulating ferroptosis.