中国中西医结合肾病杂志
中國中西醫結閤腎病雜誌
중국중서의결합신병잡지
CHINESE JOURNAL OF INTEGRATED TRADITIONAL AND WESTERN NEPHROLOGY
2014年
8期
676-681
,共6页
孟凡辰%郭兆安%于春江%孙春晓%武帅%袁婷%李悦%贾佑铎
孟凡辰%郭兆安%于春江%孫春曉%武帥%袁婷%李悅%賈祐鐸
맹범신%곽조안%우춘강%손춘효%무수%원정%리열%가우탁
芪蛭降糖胶囊%糖尿病肾病%血管内皮生长因子%细胞外基质
芪蛭降糖膠囊%糖尿病腎病%血管內皮生長因子%細胞外基質
기질강당효낭%당뇨병신병%혈관내피생장인자%세포외기질
Diabetic nephropathy%Qizhi jiangtang capsule%Vascular endothelial growth factor%Extracellular matrix
目的：观察芪蛭降糖胶囊对糖尿病肾病（diabetic nephropathy，DN）大鼠肾组织血管内皮细胞生长因子（vascu-lar endothelial growth factor，VEGF）及细胞外基质（extracellular matrix，ECM）的影响。方法：清洁级6~7周龄雄性 Wister 大鼠48只，按体重随机取40只，采用切除右肾加腹腔注射链脲佐菌素（strephozotocin，STZ）的方法制备 DN 模型。模型成功后按照血糖高低排序，两头抽取随机分为模型组、缬沙坦对照组（简称对照组）、芪蛭降糖胶囊低剂量组、芪蛭降糖胶囊高剂量组，每组10只大鼠。另取8只正常大鼠行右肾假切除术，腹腔注射等量柠檬酸缓冲液作为假手术组。成模2 d 起各组给予相应浓度和剂量的药物灌胃，分别于4周、8周观察血肌酐（Scr）、尿素氮（BUN）、血清胱抑素- C（Cystatin - C）、β2-微球蛋白（β2- MG）、24 h 尿蛋白定量（24 h TP）、尿微量白蛋白（mAlb）、尿微量白蛋白/尿肌酐（UmAlb/ Ucr）。8周后处死动物，肾组织行 HE 染色，光镜下观察肾组织病变，电镜下观察肾组织足细胞的变化。免疫组化检测 VEGF、基质金属蛋白酶9（MMP -9）、基质金属蛋白酶抑制因子-1（TIMP -1）、纤维黏连蛋白（FN）、Ⅳ型胶原蛋白（ColⅣ）的表达。结果：在各个时间点，模型组大鼠血 Scr、BUN、Cystatin - C、β2- MG，尿24 h TP、mAlb、UmAlb/ Ucr 均明显高于假手术组（P ﹤0．05），对照组明显低于模型组（P ﹤0．05），中药组显著低于对照组和模型组，且呈剂量依赖关系（P ﹤0．05~0．01）。与模型组和对照组相比，HE 染色显示芪蛭降糖胶囊能减轻 DN 大鼠肾组织及其血管病理损害。电镜观察显示，芪蛭降糖胶囊能明显减轻 DN 大鼠肾组织基底膜增厚、内皮细胞及系膜增生，减轻足细胞足突融合，且呈剂量依赖关系。免疫组化显示 VEGF、TIMP -1、FN、ColⅣ在模型组的表达较假手术组有明显增加（P ﹤0．05），对照组和低剂量组明显低于模型组（P ﹤0．05），高剂量组显著低于对照组（ P ﹤0．01）；MMP -9在假手术组表达最明显，模型组显著低于假手术组（P ﹤0．01），对照组和低剂量组高于模型组（P ﹤0．05），高剂量组显著高于对照组和低剂量组（P ﹤0．05~0．01）。结论：芪蛭降糖胶囊可以明显抑制肾组织中 VEGF 的过度合成和沉积，能够有效减轻 ECM 在肾组织中的积聚，从而改善 DN 大鼠肾组织和血管结构病理损害。
目的：觀察芪蛭降糖膠囊對糖尿病腎病（diabetic nephropathy，DN）大鼠腎組織血管內皮細胞生長因子（vascu-lar endothelial growth factor，VEGF）及細胞外基質（extracellular matrix，ECM）的影響。方法：清潔級6~7週齡雄性 Wister 大鼠48隻，按體重隨機取40隻，採用切除右腎加腹腔註射鏈脲佐菌素（strephozotocin，STZ）的方法製備 DN 模型。模型成功後按照血糖高低排序，兩頭抽取隨機分為模型組、纈沙坦對照組（簡稱對照組）、芪蛭降糖膠囊低劑量組、芪蛭降糖膠囊高劑量組，每組10隻大鼠。另取8隻正常大鼠行右腎假切除術，腹腔註射等量檸檬痠緩遲液作為假手術組。成模2 d 起各組給予相應濃度和劑量的藥物灌胃，分彆于4週、8週觀察血肌酐（Scr）、尿素氮（BUN）、血清胱抑素- C（Cystatin - C）、β2-微毬蛋白（β2- MG）、24 h 尿蛋白定量（24 h TP）、尿微量白蛋白（mAlb）、尿微量白蛋白/尿肌酐（UmAlb/ Ucr）。8週後處死動物，腎組織行 HE 染色，光鏡下觀察腎組織病變，電鏡下觀察腎組織足細胞的變化。免疫組化檢測 VEGF、基質金屬蛋白酶9（MMP -9）、基質金屬蛋白酶抑製因子-1（TIMP -1）、纖維黏連蛋白（FN）、Ⅳ型膠原蛋白（ColⅣ）的錶達。結果：在各箇時間點，模型組大鼠血 Scr、BUN、Cystatin - C、β2- MG，尿24 h TP、mAlb、UmAlb/ Ucr 均明顯高于假手術組（P ﹤0．05），對照組明顯低于模型組（P ﹤0．05），中藥組顯著低于對照組和模型組，且呈劑量依賴關繫（P ﹤0．05~0．01）。與模型組和對照組相比，HE 染色顯示芪蛭降糖膠囊能減輕 DN 大鼠腎組織及其血管病理損害。電鏡觀察顯示，芪蛭降糖膠囊能明顯減輕 DN 大鼠腎組織基底膜增厚、內皮細胞及繫膜增生，減輕足細胞足突融閤，且呈劑量依賴關繫。免疫組化顯示 VEGF、TIMP -1、FN、ColⅣ在模型組的錶達較假手術組有明顯增加（P ﹤0．05），對照組和低劑量組明顯低于模型組（P ﹤0．05），高劑量組顯著低于對照組（ P ﹤0．01）；MMP -9在假手術組錶達最明顯，模型組顯著低于假手術組（P ﹤0．01），對照組和低劑量組高于模型組（P ﹤0．05），高劑量組顯著高于對照組和低劑量組（P ﹤0．05~0．01）。結論：芪蛭降糖膠囊可以明顯抑製腎組織中 VEGF 的過度閤成和沉積，能夠有效減輕 ECM 在腎組織中的積聚，從而改善 DN 大鼠腎組織和血管結構病理損害。
목적：관찰기질강당효낭대당뇨병신병（diabetic nephropathy，DN）대서신조직혈관내피세포생장인자（vascu-lar endothelial growth factor，VEGF）급세포외기질（extracellular matrix，ECM）적영향。방법：청길급6~7주령웅성 Wister 대서48지，안체중수궤취40지，채용절제우신가복강주사련뇨좌균소（strephozotocin，STZ）적방법제비 DN 모형。모형성공후안조혈당고저배서，량두추취수궤분위모형조、힐사탄대조조（간칭대조조）、기질강당효낭저제량조、기질강당효낭고제량조，매조10지대서。령취8지정상대서행우신가절제술，복강주사등량저몽산완충액작위가수술조。성모2 d 기각조급여상응농도화제량적약물관위，분별우4주、8주관찰혈기항（Scr）、뇨소담（BUN）、혈청광억소- C（Cystatin - C）、β2-미구단백（β2- MG）、24 h 뇨단백정량（24 h TP）、뇨미량백단백（mAlb）、뇨미량백단백/뇨기항（UmAlb/ Ucr）。8주후처사동물，신조직행 HE 염색，광경하관찰신조직병변，전경하관찰신조직족세포적변화。면역조화검측 VEGF、기질금속단백매9（MMP -9）、기질금속단백매억제인자-1（TIMP -1）、섬유점련단백（FN）、Ⅳ형효원단백（ColⅣ）적표체。결과：재각개시간점，모형조대서혈 Scr、BUN、Cystatin - C、β2- MG，뇨24 h TP、mAlb、UmAlb/ Ucr 균명현고우가수술조（P ﹤0．05），대조조명현저우모형조（P ﹤0．05），중약조현저저우대조조화모형조，차정제량의뢰관계（P ﹤0．05~0．01）。여모형조화대조조상비，HE 염색현시기질강당효낭능감경 DN 대서신조직급기혈관병리손해。전경관찰현시，기질강당효낭능명현감경 DN 대서신조직기저막증후、내피세포급계막증생，감경족세포족돌융합，차정제량의뢰관계。면역조화현시 VEGF、TIMP -1、FN、ColⅣ재모형조적표체교가수술조유명현증가（P ﹤0．05），대조조화저제량조명현저우모형조（P ﹤0．05），고제량조현저저우대조조（ P ﹤0．01）；MMP -9재가수술조표체최명현，모형조현저저우가수술조（P ﹤0．01），대조조화저제량조고우모형조（P ﹤0．05），고제량조현저고우대조조화저제량조（P ﹤0．05~0．01）。결론：기질강당효낭가이명현억제신조직중 VEGF 적과도합성화침적，능구유효감경 ECM 재신조직중적적취，종이개선 DN 대서신조직화혈관결구병리손해。
Objective:To Study effects of Qizhi Jiangtang Capsule(QJC)on the expressions of VEGF and ECM in kidney tissues of DN rats. Methods:40 of the 48 male Wistar rats(6 to 7 weeks)were randomly divided into 4 groups:the sham operation group,the model group,the Valsartan group(control group),the low dosage of QJC granules group(low dosage group)and high dosage of QJC granules group(high dosage group),and each group with 10 rats. Except the rats of the sham operation group,the other rats were established DN animal model by removaling the right kidney with intraperitoneal injection of STZ. Medicine was given to each group according to the designed concentration and dosage after 2 days of the model established. Serum creatinine. blood urea nitrogen,cystatin - C,β2 - microglobulin,24 hour surinary protein quantitative,urine microalbumin and UmALB/ Ucr of DN rats were tested and recorded every 4 weeks. All animals were put to death after being treated 8 weeks. Histological change in the kidney tissues was examined by HE stain. Kidney tissue was observed under optical microscope. Kidney podocyte was observed under elec-tron microscope. VEGF,Matrix Metalloproteinase 9(MMP - 9),Matrix Metalloproteinases Inhibitory Factor1(TIMP - 1),fibronec-tin(FN)andⅣType Collagen(ColⅣ)were detected by immunohistochemical. Results:At all time points,Scr,BUN,Cystatin - C,β2 - MG,24 h TP,mALB and UmALB/ Ucr of the model group were higher than those of the sham operation group(P ﹤ 0. 05),the control group was lower than the model group(P ﹤ 0. 05),the QJC groups were higher than the sham operation group with dose de-pendent(P ﹤ 0. 05 ~ 0. 01). Compared with the model group and control group,HE staining showed that kidney and vessel pathologi-cal damage of DN rats were lessened by QJC. Observations of electron microscopy show that kidney thickening of basement membrane, endothelial cell and mesangial proliferation can obviously be relieved by QJC,podocyte foot process fusion also can be reduced with dose dependent. Immunohistochemical showed that:Expressions of VEGF,TIMP - 1,FN,Col Ⅳ in model group have increased more significantly than that of the sham operation group(P ﹤ 0. 05). Comparing these indexes,the control group and the low dosage group were significantly lower than the model group(P ﹤ 0. 05),the high dosage group was significantly lower than the low dosage group(P ﹤ 0. 01). MMP - 9 express the most obvious in the sham operation group,the model group was significantly lower than the sham operation group(P ﹤ 0. 01). The control group and the low dosage group were higher than the model group,the high dosage group was significantly higher than the control group and the low dosage group(P ﹤ 0. 05 ~ 0. 01). Conclusion:The excessive synthe-sis and deposition of VEGF in the kidney tissues can be obviously inhibited,and the ECM expression in kidney tissue can be effective-ly controlled by QJC,thus the DN rat renal pathological damage tissue and vascular structures can be improved.
