经肾动脉移植骨髓间充质干细胞对慢性肾脏病大鼠细胞程序性坏死的影响

Effects of transplantation of bone marrow mesenchymal stem cells via renal artery on necroptosis of rats with chronic kidney disease

目的探讨经肾动脉移植骨髓间充质干细胞(BMSCs)对慢性肾脏病(CKD)大鼠程序性坏死的影响。方法选取16只SD大鼠建立CKD模型并随机分为慢性肾脏病(B)组、BMSCs治疗(C)组,另随机选16只健康SD大鼠分为正常对照(N)组、培养基对照(A)组。各组接受干预后第1周末收集血、尿检测各项生化指标,处死大鼠收集肾脏,采用苏木素-伊红(HE)及马松(Masson)染色观察肾脏病理改变;采用免疫组化法观察肾脏程序性坏死相关蛋白表达程度;采用Western印迹分析程序性坏死相关蛋白的表达水平。结果与N、A组相比,B、C组肾脏损伤程度、纤维化程度、RIP1与RIP3表达量明显升高,经BMSCs治疗后,C组较B组表达量明显下降(均P<0.05);B组半胱胺酸天冬氨酸蛋白酶(caspase)8表达量明显下降,经BMSCs治疗后明显升高(均P<0.05);B组RIP1、RIP3、p-MLKL表达明显高于其余各组,经BMSCs治疗后C组各项指标较B组明显下降(均P<0.05)。结论经肾动脉移植BMSCs可抑制程序性坏死进程从而对慢性肾脏病起到较好的治疗作用。

Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells(BMSCs)via renal artery on programmed necrosis in chronic kidney disease(CKD)rats.Methods 16 SD rats were selected to establish CKD models and randomly divided into chronic kidney disease(B)group and BMSCs treatment(C)group.Another 16 healthy SD rats were randomly divided into normal control(N)group and medium control(A)group.Blood and urine were collected to test biochemical indicators at the first weekend after each group received the intervention.Rats were sacrificed in each group to collect kidneys.Hematoxylin-eosin(HE)and Masson staining were used to observe renal pathological changes.Immunohistochemical method was used to observe the expression of renal programmed necrosis related protein.Western blot was used to analyze the expression level of programmed necrosis-related proteins.Results Compared with group N and group A,the degree of kidney damage and fibrosis in group B and C were significantly increased(P<0.05);the expressions of RIP1 and RIP3 in group B and C were significantly increased(P<0.05),but after treated by BMSCs,those in group C were significantly lower than those in group B(P<0.05);the expression of caspase8 in group B was significantly decreased and it was significantly increased after treated by BMSCs.The expressions of RIP1,RIP3,and p-MLKL in group B were significantly higher than those in the other groups(P<0.05),and the expressions of those in group C were significantly lower than those in group B after treated by BMSCs(P<0.05).Conclusions Transplantation of BMSCs via renal artery could inhibit the process of programmed necrosis and play an important therapeutic effect in chronic kidney disease.