CAJ | 학술논문

目的：观察芪蛭降糖胶囊对糖尿病肾病（DN）大鼠肾脏结构和功能的影响。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佐菌素（STZ）的方法制备DN模型，造模成功后按24hTP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。另取8只大鼠行右肾假切术，腹腔注射等量柠檬酸缓冲液作为假手术组。成模2d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24h尿蛋白定量（24hUPQ）、α1微球蛋白(α1-MG)、β2-微球蛋白(β2-MG)和血糖（BS）、血清肌酐（Scr）、血清胱抑素-C（Cystatin-C）的变化；测定血浆血管紧张素Ⅱ（AngⅡ）、内皮素-1（ET-1）、一氧化氮（NO）；留取标本后处死所有动物，肾组织行HE染色、PAS染色、免疫组化染色，光镜下观察肾组织病变，PAS染色计算肾小球硬化指数（GSI）、肾间质纤维化指数（IF），电镜观察肾小球毛细血管超微结构；免疫组化检测肾组织转化生长因子（TGF）-β1的表达。结果12周后，模型组大鼠24hUPQ、α1-MG、β2-MG较假手术组显著增多（P＜0.01），3个治疗组DN大鼠的24hUPQ、α1-MG、β2-MG 较模型组明显减少（P ＜0.01），2个中药治疗组较对照组也明显减少（P＜0.05～0.01），且呈剂量依赖性；模型组大鼠BS、Scr、Cystatin-C亦较假手术组显著升高（P＜0.01），2个中药治疗组较模型组也明显降低（P＜0.05～0.01），对照组BS与模型组相比无明显变化（P＞0.05）。3个治疗组血浆AngⅡ、ET-1较模型组显著降低、NO显著升高（P＜0.05～0.01）。HE染色显示，2个中药治疗组大鼠肾组织病理损害明显减轻，PAS染色显示2个中药治疗组大鼠肾组织GSI、IF显著减轻（P＜0.01）。电镜观察显示，2个中药治疗组DN大鼠肾小球基底膜增厚和系膜增生、足细胞足突融合减轻。免疫组化检查显示2个中药治疗组DN大鼠肾组织TGF-β1的表达显著下调（P＜0.01）。结论芪蛭降糖胶囊通过抑制AngⅡ的分泌，减轻血管内皮损伤，下调TGF-β1在肾组织的表达，减轻肾小球硬化和肾间质纤维化，改善肾组织病理损害，从而对DN起到治疗作用。目的：觀察芪蛭降糖膠囊對糖尿病腎病（DN）大鼠腎髒結構和功能的影響。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佐菌素（STZ）的方法製備DN模型，造模成功後按24hTP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。另取8隻大鼠行右腎假切術，腹腔註射等量檸檬痠緩遲液作為假手術組。成模2d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24h尿蛋白定量（24hUPQ）、α1微毬蛋白(α1-MG)、β2-微毬蛋白(β2-MG)和血糖（BS）、血清肌酐（Scr）、血清胱抑素-C（Cystatin-C）的變化；測定血漿血管緊張素Ⅱ（AngⅡ）、內皮素-1（ET-1）、一氧化氮（NO）；留取標本後處死所有動物，腎組織行HE染色、PAS染色、免疫組化染色，光鏡下觀察腎組織病變，PAS染色計算腎小毬硬化指數（GSI）、腎間質纖維化指數（IF），電鏡觀察腎小毬毛細血管超微結構；免疫組化檢測腎組織轉化生長因子（TGF）-β1的錶達。結果12週後，模型組大鼠24hUPQ、α1-MG、β2-MG較假手術組顯著增多（P＜0.01），3箇治療組DN大鼠的24hUPQ、α1-MG、β2-MG 較模型組明顯減少（P ＜0.01），2箇中藥治療組較對照組也明顯減少（P＜0.05～0.01），且呈劑量依賴性；模型組大鼠BS、Scr、Cystatin-C亦較假手術組顯著升高（P＜0.01），2箇中藥治療組較模型組也明顯降低（P＜0.05～0.01），對照組BS與模型組相比無明顯變化（P＞0.05）。3箇治療組血漿AngⅡ、ET-1較模型組顯著降低、NO顯著升高（P＜0.05～0.01）。HE染色顯示，2箇中藥治療組大鼠腎組織病理損害明顯減輕，PAS染色顯示2箇中藥治療組大鼠腎組織GSI、IF顯著減輕（P＜0.01）。電鏡觀察顯示，2箇中藥治療組DN大鼠腎小毬基底膜增厚和繫膜增生、足細胞足突融閤減輕。免疫組化檢查顯示2箇中藥治療組DN大鼠腎組織TGF-β1的錶達顯著下調（P＜0.01）。結論芪蛭降糖膠囊通過抑製AngⅡ的分泌，減輕血管內皮損傷，下調TGF-β1在腎組織的錶達，減輕腎小毬硬化和腎間質纖維化，改善腎組織病理損害，從而對DN起到治療作用。
목적：관찰기질강당효낭대당뇨병신병（DN）대서신장결구화공능적영향。방법48지청길급웅성Wistar대서안체중수궤추취40지，채용절제우신가복강주사련뇨좌균소（STZ）적방법제비DN모형，조모성공후안24hTP고저，량두수궤추취분위모형조、힐사탄대조조、기질강당효낭저제량조、기질강당효낭고제량조。령취8지대서행우신가절술，복강주사등량저몽산완충액작위가수술조。성모2d기각조급여상응농도화제량적약물，급약12주시관찰DN대서24h뇨단백정량（24hUPQ）、α1미구단백(α1-MG)、β2-미구단백(β2-MG)화혈당（BS）、혈청기항（Scr）、혈청광억소-C（Cystatin-C）적변화；측정혈장혈관긴장소Ⅱ（AngⅡ）、내피소-1（ET-1）、일양화담（NO）；류취표본후처사소유동물，신조직행HE염색、PAS염색、면역조화염색，광경하관찰신조직병변，PAS염색계산신소구경화지수（GSI）、신간질섬유화지수（IF），전경관찰신소구모세혈관초미결구；면역조화검측신조직전화생장인자（TGF）-β1적표체。결과12주후，모형조대서24hUPQ、α1-MG、β2-MG교가수술조현저증다（P＜0.01），3개치료조DN대서적24hUPQ、α1-MG、β2-MG 교모형조명현감소（P ＜0.01），2개중약치료조교대조조야명현감소（P＜0.05～0.01），차정제량의뢰성；모형조대서BS、Scr、Cystatin-C역교가수술조현저승고（P＜0.01），2개중약치료조교모형조야명현강저（P＜0.05～0.01），대조조BS여모형조상비무명현변화（P＞0.05）。3개치료조혈장AngⅡ、ET-1교모형조현저강저、NO현저승고（P＜0.05～0.01）。HE염색현시，2개중약치료조대서신조직병리손해명현감경，PAS염색현시2개중약치료조대서신조직GSI、IF현저감경（P＜0.01）。전경관찰현시，2개중약치료조DN대서신소구기저막증후화계막증생、족세포족돌융합감경。면역조화검사현시2개중약치료조DN대서신조직TGF-β1적표체현저하조（P＜0.01）。결론기질강당효낭통과억제AngⅡ적분비，감경혈관내피손상，하조TGF-β1재신조직적표체，감경신소구경화화신간질섬유화，개선신조직병리손해，종이대DN기도치료작용。
[Abstrac] Objective To study effect of Qizhi Jiangtang Capsule (QJC) on renal structure and function of kedney in DN 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. 24hUPQ,α1-microglobulin (α1-MG),β2- microglobulin(β2-MG)and serum creatinine(Scr), Cystatin-C of DN rats were observed at 12th week. Ang Ⅱ、ET-1 and NO were detected ,too. The kidney tissues was stained respectively by HE,PAS and immunohistochemical.Histological change in the kidney tissues was observed by HE stain. GSI and IF were calculated by PAS stain.Glomerular capillary ultrastructure was observed by electron microscopy. The expression of TGF-β1 was detected by immunohistochemical. Results At 12th week , compared with the sham group, 24hUPQ、α1-MG、β2- <br> MG of model group were significantly raised(P < 0.01). These indicators of 3 treated groups rats were significantly reduced (P < 0.01). Compared with the control group,the above contents of 2 groups of QJC were obviously decreased (P<0.05-0.01),and dose-dependent manner. BS、Scr、Cystatin-C of the model group were also significantly raised(P<0.01)at same time.To compared with the control group,these indicators of 2 groups of QJC were obviously decreased (P < 0.05-0.01), BS of the control group was shown no significant change(P > 0.05). To compared with the control group,AngⅡ、ET-1 of 3 treated groups rats were significantly reduced(P<0.01), NO were significantly raised(P<0.01). HE staining showed that renal pathological damage were lessened by QJC. PAS stain showed that GSI、IF were significantly lessened in 2 traditional Chinese medicine treatment groups(P<0.01).Electron microscopy observations show that thickening of glomerular basement membrane and mesangium hyperplasia were reduced by QJC, podocyte foot process fusion reduced. Immunohistochemical tests show that the expression of renal tissueTGF-β1 were down-regulated by QJC(P<0.01). Conclusion Through secretion of AngⅡ was suppressed, vascular endothelial damage reduced,the expression of TGF-β1 in the renal tissues were down-regulated, structure and function of kidney tissues improved, DN is treated by QJC.