CAJ | 학술논문

目的：观察芪蛭降糖胶囊对糖尿病肾病(diabetic nephropathy,DN)大鼠肾组织骨形成蛋白-7(bone morpho-genetic protein-7,BMP-7)及转化生长因子(transforming growth factor,TGF)-β1/Smads信号传导通路的影响。方法：48只清洁级雄性Wistar大鼠按体重随机抽取40只,采用切除右肾加腹腔注射链脲菌素( streptozotocin,STZ)的方法制备DN模型,另外8只大鼠行右肾假切术。造模成功后按尿微量白蛋白( microAlbumin,mAlb)高低,两头随机抽取分为模型组、缬沙坦对照组、芪蛭降糖胶囊低剂量组、芪蛭降糖胶囊高剂量组。成模2 d起各组给予相应浓度和剂量的药物,给药12周时观察DN大鼠尿mAlb、α1微球蛋白(α1-MG)和血清肌酐( Scr)、尿素氮( BUN)。然后,处死所有动物,肾组织行HE染色、PAS染色和Masson染色,观察肾组织病理变化并半定量计算肾小管损伤指数(tubulointerstitial injury index,TII),免疫组化法检测BMP-7、TGF-β1、Smad2、Smad7在肾组织的表达。结果：给药12周后,模型组大鼠尿 mAlb、α1-MG较假手术组显著增多( P <0.01),3个治疗组DN大鼠尿mAlb、α1-MG较模型组明显减少(P<0.01),2个中药治疗组的上述4项指标亦较对照组降低(P<0.05~0.01),且呈剂量依赖性。模型组大鼠Scr、BUN亦较假手术组显著升高(P<0.01),对照组与模型组相比无明显变化(P>0.05),但2个中药治疗组较模型组明显降低(P<0.05~0.01)。 HE染色显示,3个治疗组大鼠肾组织病理损害明显减轻,2个中药治疗组大鼠肾组织病理改善比对照组更明显；PAS染色进行TII评分发现,3个治疗组大鼠TII显著低于模型组(P<0.01),2个中药治疗组TII明显低于对照组(P<0.05~0.01)；Masson染色观察发现,3个实验组大鼠肾小管间质病变明显轻于模型组,2个中药治疗组大鼠肾小管间质病变轻于对照组；免疫组化法染色并对其灰度值测定发现,3个治疗组大鼠TGF-β1、Smad2在肾组织的表达低于模型组(P<0.01),2个中药治疗组大鼠TGF-β1、Smad2在肾组织的表达低于对照组(P<0.05~0.01)；3个治疗组大鼠BMP-7、Smad7在肾脏组织的表达高于模型组(P<0.01),2个中药治疗组大鼠BMP-7、Smad7在肾组织的表达高于对照组(P<0.05~0.01)。结论：芪蛭降糖胶囊可能通过干预BMP-7/TGF-β1/Smads信号转导通路抑制了TGF-β1信号的细胞内转导,而对DN肾间质纤维化起到治疗作用。目的：觀察芪蛭降糖膠囊對糖尿病腎病(diabetic nephropathy,DN)大鼠腎組織骨形成蛋白-7(bone morpho-genetic protein-7,BMP-7)及轉化生長因子(transforming growth factor,TGF)-β1/Smads信號傳導通路的影響。方法：48隻清潔級雄性Wistar大鼠按體重隨機抽取40隻,採用切除右腎加腹腔註射鏈脲菌素( streptozotocin,STZ)的方法製備DN模型,另外8隻大鼠行右腎假切術。造模成功後按尿微量白蛋白( microAlbumin,mAlb)高低,兩頭隨機抽取分為模型組、纈沙坦對照組、芪蛭降糖膠囊低劑量組、芪蛭降糖膠囊高劑量組。成模2 d起各組給予相應濃度和劑量的藥物,給藥12週時觀察DN大鼠尿mAlb、α1微毬蛋白(α1-MG)和血清肌酐( Scr)、尿素氮( BUN)。然後,處死所有動物,腎組織行HE染色、PAS染色和Masson染色,觀察腎組織病理變化併半定量計算腎小管損傷指數(tubulointerstitial injury index,TII),免疫組化法檢測BMP-7、TGF-β1、Smad2、Smad7在腎組織的錶達。結果：給藥12週後,模型組大鼠尿 mAlb、α1-MG較假手術組顯著增多( P <0.01),3箇治療組DN大鼠尿mAlb、α1-MG較模型組明顯減少(P<0.01),2箇中藥治療組的上述4項指標亦較對照組降低(P<0.05~0.01),且呈劑量依賴性。模型組大鼠Scr、BUN亦較假手術組顯著升高(P<0.01),對照組與模型組相比無明顯變化(P>0.05),但2箇中藥治療組較模型組明顯降低(P<0.05~0.01)。 HE染色顯示,3箇治療組大鼠腎組織病理損害明顯減輕,2箇中藥治療組大鼠腎組織病理改善比對照組更明顯；PAS染色進行TII評分髮現,3箇治療組大鼠TII顯著低于模型組(P<0.01),2箇中藥治療組TII明顯低于對照組(P<0.05~0.01)；Masson染色觀察髮現,3箇實驗組大鼠腎小管間質病變明顯輕于模型組,2箇中藥治療組大鼠腎小管間質病變輕于對照組；免疫組化法染色併對其灰度值測定髮現,3箇治療組大鼠TGF-β1、Smad2在腎組織的錶達低于模型組(P<0.01),2箇中藥治療組大鼠TGF-β1、Smad2在腎組織的錶達低于對照組(P<0.05~0.01)；3箇治療組大鼠BMP-7、Smad7在腎髒組織的錶達高于模型組(P<0.01),2箇中藥治療組大鼠BMP-7、Smad7在腎組織的錶達高于對照組(P<0.05~0.01)。結論：芪蛭降糖膠囊可能通過榦預BMP-7/TGF-β1/Smads信號轉導通路抑製瞭TGF-β1信號的細胞內轉導,而對DN腎間質纖維化起到治療作用。
목적：관찰기질강당효낭대당뇨병신병(diabetic nephropathy,DN)대서신조직골형성단백-7(bone morpho-genetic protein-7,BMP-7)급전화생장인자(transforming growth factor,TGF)-β1/Smads신호전도통로적영향。방법：48지청길급웅성Wistar대서안체중수궤추취40지,채용절제우신가복강주사련뇨균소( streptozotocin,STZ)적방법제비DN모형,령외8지대서행우신가절술。조모성공후안뇨미량백단백( microAlbumin,mAlb)고저,량두수궤추취분위모형조、힐사탄대조조、기질강당효낭저제량조、기질강당효낭고제량조。성모2 d기각조급여상응농도화제량적약물,급약12주시관찰DN대서뇨mAlb、α1미구단백(α1-MG)화혈청기항( Scr)、뇨소담( BUN)。연후,처사소유동물,신조직행HE염색、PAS염색화Masson염색,관찰신조직병리변화병반정량계산신소관손상지수(tubulointerstitial injury index,TII),면역조화법검측BMP-7、TGF-β1、Smad2、Smad7재신조직적표체。결과：급약12주후,모형조대서뇨 mAlb、α1-MG교가수술조현저증다( P <0.01),3개치료조DN대서뇨mAlb、α1-MG교모형조명현감소(P<0.01),2개중약치료조적상술4항지표역교대조조강저(P<0.05~0.01),차정제량의뢰성。모형조대서Scr、BUN역교가수술조현저승고(P<0.01),대조조여모형조상비무명현변화(P>0.05),단2개중약치료조교모형조명현강저(P<0.05~0.01)。 HE염색현시,3개치료조대서신조직병리손해명현감경,2개중약치료조대서신조직병리개선비대조조경명현；PAS염색진행TII평분발현,3개치료조대서TII현저저우모형조(P<0.01),2개중약치료조TII명현저우대조조(P<0.05~0.01)；Masson염색관찰발현,3개실험조대서신소관간질병변명현경우모형조,2개중약치료조대서신소관간질병변경우대조조；면역조화법염색병대기회도치측정발현,3개치료조대서TGF-β1、Smad2재신조직적표체저우모형조(P<0.01),2개중약치료조대서TGF-β1、Smad2재신조직적표체저우대조조(P<0.05~0.01)；3개치료조대서BMP-7、Smad7재신장조직적표체고우모형조(P<0.01),2개중약치료조대서BMP-7、Smad7재신조직적표체고우대조조(P<0.05~0.01)。결론：기질강당효낭가능통과간예BMP-7/TGF-β1/Smads신호전도통로억제료TGF-β1신호적세포내전도,이대DN신간질섬유화기도치료작용。
Objective:To study the effects of Qizhi Jiangtang Capsule( QJC) on BMP-7 and TGF-β1 ,Smad2 and Smad7 expressions of renal interstitial fibrosis in diabetic nephropathy rats. Methods:48 male Wistar rats were randomly divided into 5 groups (n=8):the sham operation group,the model group,Valsartan group (control group),the low-dosage of QJC group (low dosage group) and high dosage of QJC group ( high dosage group) . Except the rats in the sham operation group,the other rats were established DN animal model by removal of the right kidney with intraperitoneal injection of STZ. Medicine was given to each group according to the designed concentration and dosage from 2 days of the model established. Urine microAlbumin(mAlb),α1 -microglobulin(α1 -MG)and serum creatinine(Scr),blood urea nitrogen(BUN) of DN rats were detect ed at 12th week. All animals were put to death af-ter 12 weeks. Histological change in the kidney tissue of DN rats was examined by HE stain,PAS stain and Masson stain. The protein expression of BMP-7,TGF-β1 ,Smad2 and Smad7 in kidney tissues were detected by ELISA respectively. Results:At 12th week,to compared with the sham group,mAlb,α1 -MG of model group were significantly raised(P<0. 01). These indicators of 3 treated groups rats were significantly reduced(P<0. 01). To compared with the control group,the above contents of 2 groups of QJC were ob-viously decreased(P<0. 05~0. 01),and dose-dependent manner. Scr、BUN of the model group were also significantly raised(P<0. 01)at same time. Scr,BUN of the control group was shown no significant change(P>0. 05). ocompared with the control group, these indicators of 2 groups of QJC were obviously decreased(P<0. 05~0. 01). HE staining showed that renal pathological damage of 3 treated groups rats were lessened. Renal pathological of 2 groups of QJC was improved more obvious than that of control group. PAS staining showed that TII of 3 treated groups was significantly lower than that of the model group,and TII of 2 groups of QJC was more lower than the control group. Tubulointerstitial was improved by Masson stain in the 2 groups of QJC. Compared with the model group, the protein expression of TGF-β1 ,Smad2 were down-regulated in the renal tissue of 3 treated groups,while the protein expression were further down-regulated by QJC. At the same time,the protein expression of BMP-7、Smad7 were up-regulated in the 3 treated groups,and that were further up-regulated by QJC. Conclusion:The renal interstitial fibrosis may be inhibitted by QJC through TGF-β1/Smads signaling pathway was interferenced in the renal tissues of DN rats.