中西医结合学报
中西醫結閤學報
중서의결합학보
JOURNAL OF CHINESE INTEGRATIVE MDEICINE
2011年
2期
201-208
,共8页
陶艳艳%王清兰%袁继丽%沈丽%刘成海
陶豔豔%王清蘭%袁繼麗%瀋麗%劉成海
도염염%왕청란%원계려%침려%류성해
维生素E%肾间质纤维化%升汞%脂质过氧化反应%动物实验%大鼠
維生素E%腎間質纖維化%升汞%脂質過氧化反應%動物實驗%大鼠
유생소E%신간질섬유화%승홍%지질과양화반응%동물실험%대서
vitamin E%renal interstitial fibrosis%mercuric chloride%lipid peroxidation%animal experimentation%rats
目的:观察维生素E对氯化汞诱导的大鼠肾间质纤维化的作用并探讨其抗氧化机制方法:32只大鼠随机分为正常组、模型组与维生素E组.除正常组外,其余大鼠以8 mg/kg氯化汞(mercuric chloride,HgCl2)灌胃,每天1次,共9周.维生素E组同时以维生素E 100 mg/kg灌胃,正常组与模型组灌服等量生理盐水.9周后处死大鼠,盐酸水解法检测肾组织羟脯氨酸(hydroxyproline,Hyp)含量,苏木精伊红染色、Masson染色与过碘酸六胺银染色观察肾组织病理形态与胶原沉积;试剂盒方法检测肾组织过氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性与谷胱甘肽(glutathione,GSH)和丙二醛(malondialdehyde,MDA)含量;蛋白印迹法检测核因子κB(nuclear factor-κB,NF-κB)信号途径的κB抑制蛋白(inhibitor κB,IκB)、磷酸化IκB(phospho-IκB,p-IκB)以及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达;蛋白印迹法与免疫荧光法观察α平滑肌肌动蛋白(a-smooth muscle actin,α-SMA)的表达.结果:与正常组比较,肾组织Hyp含量、Masson染色与过碘酸六胺银染色以及肾组织α-SMA的变化证实肾间质纤维化模型造模成功;GSH-Px活性与GSH、MDA含量变化提示肾间质纤维化模型存在脂质过氧化损伤.与模型组比较,维生素E组大鼠肾组织Hyp含量降低(P<0.01),肾间质纤维化减轻,GSH与MDA含量降低(P<0.01);维生素E组p-IκB、TNF-α、α-SMA蛋白表达明显降低,各组中IκB蛋白表达无明显变化.结论:维生素E抗氯化汞诱导大鼠肾间质纤维化的作用机制在于抗脂质过氧化损伤,与抑制NF-κB信号通路及细胞外基质产生细胞活化有关.
目的:觀察維生素E對氯化汞誘導的大鼠腎間質纖維化的作用併探討其抗氧化機製方法:32隻大鼠隨機分為正常組、模型組與維生素E組.除正常組外,其餘大鼠以8 mg/kg氯化汞(mercuric chloride,HgCl2)灌胃,每天1次,共9週.維生素E組同時以維生素E 100 mg/kg灌胃,正常組與模型組灌服等量生理鹽水.9週後處死大鼠,鹽痠水解法檢測腎組織羥脯氨痠(hydroxyproline,Hyp)含量,囌木精伊紅染色、Masson染色與過碘痠六胺銀染色觀察腎組織病理形態與膠原沉積;試劑盒方法檢測腎組織過氧化物歧化酶(superoxide dismutase,SOD)和穀胱甘肽過氧化物酶(glutathione peroxidase,GSH-Px)活性與穀胱甘肽(glutathione,GSH)和丙二醛(malondialdehyde,MDA)含量;蛋白印跡法檢測覈因子κB(nuclear factor-κB,NF-κB)信號途徑的κB抑製蛋白(inhibitor κB,IκB)、燐痠化IκB(phospho-IκB,p-IκB)以及腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)的錶達;蛋白印跡法與免疫熒光法觀察α平滑肌肌動蛋白(a-smooth muscle actin,α-SMA)的錶達.結果:與正常組比較,腎組織Hyp含量、Masson染色與過碘痠六胺銀染色以及腎組織α-SMA的變化證實腎間質纖維化模型造模成功;GSH-Px活性與GSH、MDA含量變化提示腎間質纖維化模型存在脂質過氧化損傷.與模型組比較,維生素E組大鼠腎組織Hyp含量降低(P<0.01),腎間質纖維化減輕,GSH與MDA含量降低(P<0.01);維生素E組p-IκB、TNF-α、α-SMA蛋白錶達明顯降低,各組中IκB蛋白錶達無明顯變化.結論:維生素E抗氯化汞誘導大鼠腎間質纖維化的作用機製在于抗脂質過氧化損傷,與抑製NF-κB信號通路及細胞外基質產生細胞活化有關.
목적:관찰유생소E대록화홍유도적대서신간질섬유화적작용병탐토기항양화궤제방법:32지대서수궤분위정상조、모형조여유생소E조.제정상조외,기여대서이8 mg/kg록화홍(mercuric chloride,HgCl2)관위,매천1차,공9주.유생소E조동시이유생소E 100 mg/kg관위,정상조여모형조관복등량생리염수.9주후처사대서,염산수해법검측신조직간포안산(hydroxyproline,Hyp)함량,소목정이홍염색、Masson염색여과전산륙알은염색관찰신조직병리형태여효원침적;시제합방법검측신조직과양화물기화매(superoxide dismutase,SOD)화곡광감태과양화물매(glutathione peroxidase,GSH-Px)활성여곡광감태(glutathione,GSH)화병이철(malondialdehyde,MDA)함량;단백인적법검측핵인자κB(nuclear factor-κB,NF-κB)신호도경적κB억제단백(inhibitor κB,IκB)、린산화IκB(phospho-IκB,p-IκB)이급종류배사인자-α(tumor necrosis factor-α,TNF-α)적표체;단백인적법여면역형광법관찰α평활기기동단백(a-smooth muscle actin,α-SMA)적표체.결과:여정상조비교,신조직Hyp함량、Masson염색여과전산륙알은염색이급신조직α-SMA적변화증실신간질섬유화모형조모성공;GSH-Px활성여GSH、MDA함량변화제시신간질섬유화모형존재지질과양화손상.여모형조비교,유생소E조대서신조직Hyp함량강저(P<0.01),신간질섬유화감경,GSH여MDA함량강저(P<0.01);유생소E조p-IκB、TNF-α、α-SMA단백표체명현강저,각조중IκB단백표체무명현변화.결론:유생소E항록화홍유도대서신간질섬유화적작용궤제재우항지질과양화손상,여억제NF-κB신호통로급세포외기질산생세포활화유관.
Objective: To observe the effects of vitamin E(Vit E)on mercuric chloride(HgCl2)-induced renal interstitial fibrosis(RIF)in rats and discuss its antioxidative mechanism.Methods: A total of 32 Sprague-Dawley rats were randomly assigned to three groups: normal group,model group and Vit E group.RIF was induced by oral administration of HgCl2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Rats in Vit E group were administered with Vit E capsule at 100 mg/kg body weight,and rats in normal and model groups were treated with normal saline.At the end of the 9th week,rats were sacrificed and renal hydroxyproline(Hyp)content was assayed with Jamall's method and collagen deposition was visualized by using hematoxylin and eosin(HE),Masson' s trichrome and periodic acid-silver methenamine(PASM)staining.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and contents of glutathione(GSH)and malondialdehyde(MDA)in kidney tissue were tested with commercial kits.The expressions of nuclear factor-κB(NF-κB),inhibitor-κB(IκB),phospho-IκB(p-IκB)and tumor necrosis factor-α(TNF-α)were determined by Western blot.The expression of α-smooth muscle actin(α-SMA)was assayed by Western blot and immunofluorescent staining.Results: Renal Hyp content,HE,Masson's trichrome and PASM staining results and α-SMA expression confirmed development of HgCl2-induced RIF in rats.Oxidative stress markers GSH,GSH-Px and MDA confirmed oxidative stress in RIF rats.Compared with model rats,rats in Vit E group had lower kidney Hyp content(P<0.01).GSH and MDA contents decreased significantly in Vit E group compared with model group(P<0.01).The expressions of NF-κB and IκB had no significant difference among all groups(P>0.05).In Vit E group,the expressions of p-IκB and TNF-α decreased significantly compared with model group(P<0.01).The expression of α-SMA in Vit E group was also decreased significantly compared with model group(P<0.01).Conclusion: Vit E has a protective effect on experimental RIF induced by HgCl2 in rats and it is related to inhibition of lipid peroxidation,which involves blocking of NF-κB signaling pathway and the activation of cells producing extracellular matrix.
