摘要
目的:探讨氯化汞(HgCl2)灌胃诱导大鼠肾间质纤维化模型的脂质过氧化损伤病理机制。方法:以8 mg/kgHgCl2水溶液灌胃9周,设1周、2周、4周、6周和9周5个观察时间点。试剂盒方法检测肾功能(Scr、BUN)和脂质过氧化情况(GSH、GSH-Px、MDA、SOD),盐酸水解法检测肾组织羟脯氨酸(Hyp)含量,HE染色和Masson染色观察肾组织病理形态,免疫组化观察肾组织金属硫蛋白(MT)的表达和分布,免疫印记法观察核因子-κB(NF-κB)信号途径的激活(IκB、P-IκB、TNF-α)和α-平滑肌肌动蛋白(α-SMA)等蛋白的表达。结果:与正常组相比,9周模型大鼠血清Scr、BUN含量增高(Scr,P<0.05;BUN,P<0.01),肾组织GSH含量、GSH-Px活力明显降低(GSH,P<0.05;GSH-Px,P<0.01),MDA含量升高(P<0.05),SOD活力各组无差异(P>0.05);肾组织Hyp含量逐渐升高(P<0.05)。模型大鼠肾组织炎性浸润,肾间质胶原沉积增多,间质增宽,肾小管萎缩,表现出较为典型的肾间质纤维化。模型大鼠肾组织MT表达逐渐减少;IκB表达无差异(P>0.05),P-IκB、TNF-α蛋白表达均明显增加(P-IκB,P<0.05;TNF-α,P<0.01),α-SMA表达增加(P<0.01)。结论:HgCl2诱导大鼠肾间质纤维化病变机制在于肾组织脂质过氧化损伤,从而诱导NF-κB信号途径的活化和肌成纤维细胞活化,最终造成肾间质纤维化。
Objective：To explore the Mechanism of Lipid Peroxidation in Renal Interstitial Fibrosis in Rats Intoxicated by Mercuric Chloride. Methods-Rats were orally administrated with 8 mg/kg mercuric chloride （HgCl2）for 9 weeks. The dynamic changes were observed at 5 time points （1 w,2 w,4 w,6 w,9 w）. Renal function （Cr and BUN） and renal lipid peroxidation injury （GSH, GSH- Px, MDA and SOD） were tested with respective kits. Hydroxyproline （Hyp） content of kidney was assayed with hydrochloric acid hydrolysis; renal pathological changes were observed with HE and Masson staining. The expression and distribution of metallothionein （MT） was observed with immunohistochemistry. The activation of nuclear factor- kappa B（ NF- κB） signal path- way which including the expression of inhibitor- κB （IκB） protein, phospho- IκB （P- IκB） protein, tumor necrosis factor- α （TNF - α） protein and α- smooth muscle actin （α - SMA ） protein were observed with western blot. Results： Contrasted by normal rats and at the end of the 9th week, model rats had serum Cr and BUN increased （Cr, P 〈 0.05; BUN, P 〈 0.01 ）, renal GSH content and GSH- Px activity decreased （GSH, P〈0.05;GSH- Px, P〈0.01 ）, renal MDA content（P〈0.05） increased,but no difference in SOD activity（ P 〉 0.05）. In model rats, renal Hyp content also was increased （ P 〈 0.05）. During the formation of the model,inflammatory cells infiltration and increased ECM deposition were observed in renal interstitial space, renal interstitial space widened and some renal tubules atrophied, which were all in accordance with typical renal interstitial fibrosis. The expression of MT was found to be decreased gradually in model kidney, while the expression of IκB protein had no difference（P〉0.05） and that of P - IκB and TNF- α（P - IκB, P 〈 0.05 ; TNF - α, P 〈 0.01 ） increased. And the expression of α - SMA protein was also increased （ P 〈 0.01 ）. Conclusion： The mechanism of renal interstitial fibrosis in rats intoxicated by mercuric chloride was renal lipid peroxidation injury, which induced the activation of NF- κB signaling pathway and the activation of myofibroblast. And renal interstitial fibrosis formed as a result.
出处
《中国中西医结合肾病杂志》
2008年第5期403-407,I0007,共6页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
上海市科委重点基础研究项目(No.04JC14069)
教育部新世纪优秀人才计划资助项目(No.NCET-04-0437)
上海市教委高校E-研究院建设计划资助项目(No.E03008)
上海市重点学科建设资助项目(No.Y0302)
关键词
氯化汞
肾间质纤维化
脂质过氧化
金属硫蛋白
核因子-ΚB
Mercuric chloride
Renal interstitial fibrosis
Lipid peroxidationMetallothionein
Nuclear factor- kappa B
