硫化氢对甲基胍加重残余肾功能损害的保护作用

Effect of hydrogen sulfide on methylguanidine aggravating residual renal damage

目的检测残余肾功能相关指标,明确硫化氢(H2S)对甲基胍加重肾功能损害的保护作用。方法大鼠随机分为假手术(SH)组、和5/6肾切除肾衰模型(MD)组、甲基胍(MH)组及甲基胍+硫化氢(MH+H2S)组,检测各组血清尿素氮、肌酐,观察肾脏组织的病理改变,免疫组化法检测肾组织内TGF、CTGF、FN的表达水平,TUNEL观察细胞的凋亡,Western blot检测自噬相关蛋白Atg3和Beclin的表达水平。结果 MD组、MH组及MH+H2S组较假手术组血清尿素氮、肌酐显著升高(P<0.05),肾小球体积增大、系膜细胞及基质明显增生,免疫组化染色显示TGF、CTGF、FN阳性面积表达显著增强(P<0.05),细胞凋亡率显著增加(P<0.05),Western blot结果显示,Atg3和Beclin的表达水平显著增强(P<0.05)。加入甲基胍后血清尿素氮、肌酐较模型组明显升高(P<0.05),肾小球体积较模型组明显增大、系膜细胞及基质较模型组增生严重,免疫组化染色显示TGF、CTGF、FN均较模型组表达增强(P<0.05),细胞凋亡率较模型组显著增加(P<0.05),Atg3和Beclin的表达水平显著增强(P<0.05)。而加入H2S后血清尿素氮、肌酐明显下降(P<0.05),免疫组化显示TGF、CTGT、FN表达减弱(P<0.05),细胞凋亡率显著降低(P<0.05),Beclin的表达水平进一步增强(P<0.05)。结论甲基胍可能是残余肾功能进一步下降的关键因素,H2S可能通过自噬激活逆转甲基胍的肾毒性作用。

Objective To detect residual renal function related indicators and clarify the protective effect of hydrogen sulfide on methylguanidine in aggravating renal function impairment.Methods Rats were randomly divided into sham(SH)group,5/6 nephrectomy kidney failure model(MD)group,methylguanidine(MH)group and methylguanidine+hydrogen sulfide(MH+H2 S)group.The serum urea nitrogen and creatinine were detected in each group.The pathological changes of renal tissue were observed.The expression of TGF,CTGF and FN in renal tissue was detected by immunohistochemistry.TUNEL apoptosis was observed,and the expression levels of autophagy related proteins Atg3 and Beclin were detected by Western blot.Results The serum urea nitrogen and creatinine increased significantly in the MD,MH and MH+H2 S groups(P<0.05).The glomerular volume increased,the mesangial cells and the matrix proliferated.The immunohistochemical staining showed that the positive area of TGF,CTGF and FN increased significantly(P<0.05),and the apoptosis rate increased significantly(P<0.05).The results showed that expression level of Atg3 and Beclin increased significantly(P<0.05).After the addition of methylguanidine,the serum urea nitrogen and creatinine increased significantly(P<0.05).The glomerular volume increased significantly compared with the model group,and the mesangial cells and the matrix were more severe than the model group.The immunohistochemical staining showed that the expression of TGF,CTGF and FN increased compared with the model group(P<0.05),and the apoptosis rate was significantly increased(P<0.05),the expression levels of Atg3 and Beclin were significantly increased(P<0.05).After the addition of H2S,serum urea nitrogen and creatinine decreased significantly(P<0.05).Immunohistochemistry showed that the expression of TGF,CTGT and FN decreased(P<0.05),the apoptosis rate decreased significantly(P<0.05),and the Beclin expression level was further enhanced(P<0.05).Conclusion Methylguanidine may be a key factor in the further decline of residual renal function.Hydrogen sulfide may reverse the renal toxicity of methylguanidine through autophagy.