CAJ | 학술논문

目的：探讨帕立骨化醇（ paricalcitol，P）对糖尿病肾病（ DN）肾小管间质纤维化的干预作用及可能机制。方法：大鼠禁食后，采用单次无菌腹腔注射链脲佐菌素建立DN模型。将DN大鼠随机分为：（1）帕立骨化醇干预组（P组）：帕立骨化醇溶于丙二醇中，于造模成功后第2天以0.4μg／kg的剂量腹腔注射，每周3次；（2）糖尿病肾病组（ D组）：给予等体积的丙二醇腹腔注射。设置正常对照组（ C组）。帕立骨化醇连续干预12周后，测血、尿生化指标；进行肾脏病理学检查；利用免疫组化及Western blotting检测肾组织TGF-β1、Wnt-4、β-catenin及Klotho蛋白的表达；并进行指标间的相关分析。结果：（1）与C组比较，D组大鼠SCr、BUN及24 h尿蛋白水平均升高，而P组均较D组降低（P＜0.05）。（2）与C组比较，D组大鼠肾小管间质纤维化面积增加，而P组较D组减小（P＜0.05）。（3） D组大鼠肾组织Klotho蛋白表达低于C组，而P组高于D组（P＜0.05）；与C组比较，D组大鼠肾组织TGF-β1、Wnt-4及β-catenin蛋白表达增加，而P组表达均较D组减少（P＜0.05）。（4） Klotho与纤维化面积、TGF-β1、Wnt-4及β-catenin均呈负相关（ P＜0.05）。结论：帕立骨化醇可抑制DN大鼠肾小管间质纤维化，其作用可能与增加肾组织Klotho表达，抑制Wnt／β-catenin信号通路激活，同时减少TGF-β1合成相关。
목적：탐토파립골화순（ paricalcitol，P）대당뇨병신병（ DN）신소관간질섬유화적간예작용급가능궤제。방법：대서금식후，채용단차무균복강주사련뇨좌균소건립DN모형。장DN대서수궤분위：（1）파립골화순간예조（P조）：파립골화순용우병이순중，우조모성공후제2천이0.4μg／kg적제량복강주사，매주3차；（2）당뇨병신병조（ D조）：급여등체적적병이순복강주사。설치정상대조조（ C조）。파립골화순련속간예12주후，측혈、뇨생화지표；진행신장병이학검사；이용면역조화급Western blotting검측신조직TGF-β1、Wnt-4、β-catenin급Klotho단백적표체；병진행지표간적상관분석。결과：（1）여C조비교，D조대서SCr、BUN급24 h뇨단백수평균승고，이P조균교D조강저（P＜0.05）。（2）여C조비교，D조대서신소관간질섬유화면적증가，이P조교D조감소（P＜0.05）。（3） D조대서신조직Klotho단백표체저우C조，이P조고우D조（P＜0.05）；여C조비교，D조대서신조직TGF-β1、Wnt-4급β-catenin단백표체증가，이P조표체균교D조감소（P＜0.05）。（4） Klotho여섬유화면적、TGF-β1、Wnt-4급β-catenin균정부상관（ P＜0.05）。결론：파립골화순가억제DN대서신소관간질섬유화，기작용가능여증가신조직Klotho표체，억제Wnt／β-catenin신호통로격활，동시감소TGF-β1합성상관。
[ ABSTRACT] AIM:To investigate the effect of paricalcitol ( P) on renal tubulointerstitial fibrosis and the under-lying mechanisms in diabetic nephropathy ( DN) .METHODS:DN rat model was induced by a single intraperitoneal in-jection of streptozotocin after fasting.The animals were randomly divided into 2 groups: the DN rats in paricalcitol-inter-vened group ( group P) were injected intraperitoneally with paricalcitol dissolved in propylene glycol after the day when the model was induced successfully at a dose of 0.4μg/kg (3 times a week);the DN rats in DN group ( group D) were given isopyknic propylene glycol.Normal control group ( group C) was also set up.The samples of blood, urine and renal tissue were collected after intervention of paricalcitol for 12 weeks.The biochemical indexes were measured.The renal tissues were used for pathologic observation and determining the expression of transforming growth factor-β1 (TGF-β1), Wnt-4,β-catenin and Klotho by immunohistochemistry and Western blotting.In addition, the correlation among the above indexes was analyzed.RESULTS:(1) Scr, BUN and 24 h urine protein increased significantly in group D compared with group C, while decreased in group P compared with group D ( P<0.05 ) .( 2 ) The area of renal tubulointerstitial fibrosis in-creased in group D compared with group C, while decreased in group P compared with group D (P<0.05).(3) The ex-pression of Klotho decreased, while the expression of TGF-β1, Wnt-4 and β-catenin increased in group D compared with group C (P<0.05).Compared with group D, the expression of Klotho increased, while the expression of TGF-β1, Wnt-4 andβ-catenin decreased in group P (P<0.05).(4) The expression of Klotho was negatively correlated with the fibrosis area, TGF-β1, Wnt-4 andβ-catenin (P<0.05).CONCLUSION:Paricalcitol inhibits renal tubulointerstitial fibrosis in DN by promoting the expression of renal Klotho, and inhibiting Wnt/β-catenin signaling pathway activation and TGF-β1 synthesis.