摘要
目的探讨匹伐他汀(pitavastatin,PIT)对糖尿病肾脏疾病大鼠刺猬信号通路及肾脏纤维化的影响。方法20只斯泼累格-多雷大鼠按照完全随机法分为空白组、模型组、PIT-L组(0.1 mg/kg PIT)、PIT-M组(0.2 mg/kg)、PIT-H组(0.4 mg/kg),每组4只,除空白组外,其余各组大鼠均建立糖尿病肾脏疾病模型,模型构建成功后根据各组大鼠PIT给药剂量行灌胃处理,模型组和空白组正常喂养(灌胃生理盐水),持续灌胃12周后,采集各组大鼠的24 h尿液、血清及肾脏组织;测定各组大鼠血糖值、肾脏指数、尿蛋白、血肌酐、血尿素氮含量;以苏木精-伊红染色(hematoxylin-eosin staining,HE)检测各组大鼠肾脏组织病理变化:以马松染色(Masson's trichrome staining,Masson)观察大鼠肾脏纤维化程度;以蛋白免疫印迹法检测各组大鼠肾脏组织中肾脏纤维化相关蛋白及Hedgehog信号通路相关蛋白的表达。结果与空白组相比,模型组大鼠肾小球体积增大,肾小管间质局部炎症细胞浸润,肾小管上皮细胞脱落,小管扩张,系膜区增宽,肾脏纤维化面积占比较多(t=12.754,P=0.000,P<0.05);与模型组相比,PIT-L、PIT-M、PIT-H组肾脏组织形态学逐渐改善,且剂量增加,肾组织病变缓解更加明显,肾脏纤维化面积比逐渐降低(t=2.505、2.983、3.232;P=0.046、0.025、0.018)(P<0.05)。与空白组相比,模型组大鼠血糖值、肾脏指数、尿蛋白、血肌酐、血尿素氮水平、音猬因子(sonic hedgehog,Shh)、补缀同源物1(protein patched homolog 1,Ptch-1)、平滑蛋白(smoothened protein,Smo)、胶质瘤相关癌基因同源蛋白1(glioma associated oncogene homolog 1,Gli1)、转化生长因子β(transforming growth factor-β,TGF-β)1、α-平滑肌动蛋白(α-smooth muscle actin,α-SMA)、胶原蛋白1(collagen-1,Col-1)蛋白表达水平显著升高(t分别为10.816、10.568、10.655、6.573、6.572、12.691、8.943、9.481、10.236、11.449、9.911、11.359;均P<0.05);与模型组相比,PIT-L、PIT-M、PIT-H组血糖值、肾脏指数、尿蛋白、血肌酐、血尿素氮水平、Shh、Ptch-1、Smo、Gli1、TGF-β1、α-SMA、Col-1蛋白表达水平均显著降低(t分别为3.826、5.781、7.532;2.723、5.176、8.601;2.803、5.225、7.106;2.624、4.418、5.687;2.688、4.029、5.508;2.603、5.340、8.709;2.517、4.041、6.665;2.458、4.469、7.242;2.475、4.835、9.113;2.505、5.427、9.443;2.867、4.774、8.663;2.684、4.618、8.836;均P<0.05),且PIT-H组以上指标变化最明显。各组之间以上指标对比差异具有统计学意义,其中血糖值、肾脏指数、尿蛋白、血肌酐、血尿素氮水平F分别为47.493、48.917、42.917、24.676、24.174,均P<0.0001;肾脏纤维化面积F为22.907,P<0.0001;Shh、Ptch-1、Smo、Gli1蛋白表达水平F分别为54.365、29.635、36.004、45.068,均P<0.0001;TGF-β1、α-SMA、Col-1蛋白表达水平F分别为55.060、42.340、54.532,均P<0.0001。结论PIT可缓解糖尿病肾脏疾病大鼠肾脏纤维化,可能是通过抑制刺猬信号通路实现的。
Objective To investigate the effects of pitavastatin(PIT)on the Hedgehog signal⁃ing pathway and renal fibrosis in diabetic kidney disease rats.Methods Twenty Sprague-Dawley(SD)rats were randomly divided into blank group,model group,low-dose PIT group(0.1 mg/kg PIT,PITL),medium-dose PIT group(0.2 mg/kg PIT,PIT-M),and high-dose PIT group(0.4 mg/kg PIT,PIT-H),with four rats in each group.Except for the blank group,an in vivo diabetic kidney disease model was established in the remaining groups,followed by intragastric administration of PIT(PIT groups),or normal saline(blank and model groups)for 12 weeks.Rat 24-h urine,serum and kidney tis⁃sue were then collected to examine random blood glucose(RBG),renal index,urinary protein,serum creatinine(Scr)and blood urea nitrogen(BUN).Hematoxylin and eosin(H&E)staining was used to detect the pathological changes in rat kidney tissues.Masson staining was used to observe the degree of renal fibrosis.Western blot was used to detect protein expressions of fibrosis-related and the Hedgehog signaling pathway-related proteins in rat kidney tissues.Results Compared with the blank control,rats in the model group exhibited significantly increased glomerular volume,focal inflammatory cell infiltra⁃tion in the renal tubulointerstitium,tubular epithelial cell desquamation,tubular dilatation,widened me⁃sangial areas,and a larger area of renal fibrosis(t=12.754,P<0.001).Compared with the model group,the renal histology gradually improved in the PIT-L,PIT-M and PIT-H groups,showing a dose-dependent improvement in the renal pathology and renal fibrotic area(t=2.505,2.983,and 3.232;P=0.046,0.025,and 0.018;respectively).Compared with the blank control,rats in the model group showed significantly elevated RBG,renal index,urinary protein,Scr,BUN,and protein expressions of Shh,Ptch-1,Smo,Gli1,TGF-β1,α-SMA,and Col-1(t=10.816,10.568,10.655,6.573,6.572,12.691,8.943,9.481,10.236,11.449,9.911,and 11.359).Compared with the model group,rats in the PIT-L,PIT-M and PIT-H groups showed significantly reduced RBG,renal index,urinary protein,serum Cr,BUN,and protein expressions of Shh,Ptch-1,Smo,Gli1,TGF-β1,α-SMA,and Col-1(PIT-L:t=3.826,5.781,7.532,2.723,5.176,8.601,2.803,5.225,7.106,2.624,4.418 and 5.687;PIT-M:t=2.688,4.029,5.508,2.603,5.340,8.709,2.517,4.041,6.665,2.458,4.469 and 7.242;PIT-H:t=2.475,4.835,9.113,2.505,5.427,9.443,2.867,4.774,8.663,2.684,4.618 and 8.836).Notably,the most pronounced changes in these indicators were seen in the PIT-H group.Differences among these groups were statistically significant,with F values of RBG,renal index,urinary protein,serum Cr,BUN,renal fibrotic area,and protein expressions of Shh,Ptch-1,Smo,Gli1,TGF-β1,α-SMA,and Col-1 of 47.493,48.917,42.917,24.676,24.174,22.907,54.365,29.635,36.004,45.068,55.060,42.340 and 54.532,respectively(all P<0.0001).Conclusion PIT can alleviate renal fibrosis in diabetic nephropathy rats by inhibiting the Hedgehog signaling pathway.
作者
李晓璐
许焱
Li Xiao-lu;Xu Yan(Department of Nephrology,Emergency General Hospital,Beijing 102699,China)
出处
《临床肾脏病杂志》
2025年第9期790-797,共8页
Journal of Clinical Nephrology
关键词
糖尿病肾脏疾病
刺猬信号通路
肾纤维化
匹伐他汀
Diabetic kidney disease
Hedgehog signaling pathway
Renal fibrosis
Pitavastatin
