1,25-二羟维生素D3通过调节性T细胞延缓肾间质纤维化的作用研究

Effect of 1,25-dihydroxyvitamin D3 delaying renal interstitial fibrosis by T regulatory cell

目的研究1,25-二羟维生素D3[1,25(OH)2D3]对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响。方法采用随机数字表法将54只wistar大鼠分为假手术组(A组)、UUO组(B组)和1,25(OH)2D3治疗组(C组),每组18只。C组灌胃1,25(OH)2D33 ng/(d·100 g),A组和B组灌胃等量花生油。B组和C组左侧输尿管结扎,A组输尿管只游离不结扎,每组术后第1、3、7天分批处死6只大鼠,并取留取左侧肾脏。常规检测血肌酐水平,苏木精-伊红(HE)染色观察肾脏病理形态,免疫组化法检测大鼠肾脏平滑肌动蛋白(a-SMA)、叉头P3蛋白(FOXP3),Western blot法测磷酸化腺苷酸活化蛋白激酶(p-AMPK)。结果术后第1、3、7天B组血肌酐、a-SMA水平均高于A组,C组血清肌酐、a-SMA水平均低于B组(均P<0.05)。A组大鼠肾脏组织形态正常,B组大鼠肾脏组织随梗阻时间延长,可见肾小管扩张及间质水肿,而C组较B组肾小管变性减轻。术后第1、3、7天B组p-AMPK、FOXP3水平均低于A组,C组p-AMPK、FOXP3水平均高于B组(均P<0.05)。结论1,25(OH)2D3可延缓肾间质纤维化,其机制可能与激活AMPK进而促进调节性T细胞的活化有关。

Objective To study the effect of 1,25-dihydroxyvitamin D3[1,25(OH)2D3]on renal interstitial fibrosis in rats with unilateral ureteral obstruction(UUO).Methods According to random number table method,54 wistar rats were divided into sham operation group(group A),UUO group(group B)and 1,25(OH)2D3 treatment group(group C),with 18 rats in each group.Group C was gavaged 3 ng/(d·100 g)of 1,25(OH)2D3 every day.Group A and group B were gavaged the same amount of peanut oil.The left ureter ligation was performed in group B and group C.Group A was treated with ureteral free,but not ligated.On the 1st,3rd and 7th day after operation,6 rats were killed in batches,and the left kidney was retained.Serum creatinine were measured in each group.Renal pathological changes were observed by hematoxylin-eosin(HE)staining.The expression of a-smooth muscle actin(a-SMA)and forkhead P3 protein(FOXP3)in the kidney was detected by immunohistochemistry.Western blot was used to detect the protein expression of phosphorylated adenosine monophosphate activated protein kinase(p-AMPK).Results On the 1st,3rd and 7th day after operation,the serum creatinine,a-SMA levels of group B was higher than those of group A,and the serum creatinine,a-SMA levels of group C was lower than those of group B(all P<0.05).The renal tissue morphology of group A was normal,while the renal tissue of group B was prolonged with the obstruction time,showing renal tubules dilatation and interstitial edema,while the tubular degeneration of group C was reduced compared with that of group B.On the 1st,3rd and 7th day after operation,p-AMPK and FOXP3 levels in group B were lower than those in group A,p-AMPK and FOXP3 levels in group C were higher than those in group B(all P<0.05).Conclusion 1,25(OH)2D3 can delay renal interstitial fibrosis,and its mechanism may be related to the activation of AMPK to promote the activation of regulatory T cells.