自拟调气汤对顺铂诱导的大鼠肾损伤的影响及其机制

Effect of self-made Tiaoqi Decoction on cisplatin induced renal injury in rats and its mechanism

目的:研究自拟调气汤对顺铂诱导的肾损伤大鼠的影响及其机制。方法:选取90只SD雄性大鼠,随机分为正常对照组,模型组,调气汤低、中、高剂量组及阳性对照组,每组15只;除正常对照组外,其余组采用尾静脉注射顺铂(2.5 mg/kg)的方法建立肾损伤大鼠模型,造模完成后,调气汤低、中、高剂量组分别予以2、4、8 mL/kg调气汤灌胃,阳性对照组予以1 mg/kg氨磷汀腹腔注射,正常对照组及模型组给予生理盐水(2 mL/kg)灌胃,1次/d,连续60 d;比色法检测血清尿素氮、肌酐水平;ELISA法测定肾组织肿瘤坏死因子-α(TNF-α)、IL-6、IL-1β含量,以及脾脏淋巴细胞分泌的TNF-α、IL-17、基质金属蛋白酶-9(MMP-9)水平;分别用比色法和显色法检测肾组织超氧化物歧化酶(SOD)活性及丙二醛水平;HE染色检测肾组织病理变化,蛋白免疫印迹法检测肾组织中Toll样受体4(TLR4)、NF-κB蛋白表达水平。结果:与模型组比较,调气汤低、中、高剂量组血清尿素氮、肌酐水平,肾组织TNF-α、IL-6、IL-1β及丙二醛含量,肾组织TLR4及NF-κB蛋白表达水平,脾脏TNF-α、IL-17、MMP-9水平显著降低(P<0.05),肾组织SOD活性显著升高(P<0.05),肾组织病理明显改善,且随着调气汤剂量增加,各项指标呈剂量依赖性变化。结论:自拟调气汤可能通过抑制TLR4/NF-κB通路,进而显著改善顺铂诱导的大鼠肾损伤。

Objective:To investigate effect of self-made Tiaoqi Decoction on cisplatin induced renal injury in rats and its potential mechanism.Methods:Ninety male Sprague-Dawley rats were randomly divided into normal control group,model group,low,medium and high dose groups of Tiaoqi Decoction,and positive control group(n=15 for each group).Cisplatin(2.5 mg/kg)was injected via the rat′s tail vein to induce kidney injury.The low,medium and high dose groups of Tiaoqi Decoction were administrated to rats at the doses of 2,4,and 8 mL/kg by gavage.The positive control group was intraperitoneally injected with amifostine at a dose of 1 mg/kg,and the control group and model group rats were administrated with the normal saline(2 mL/kg)by gavage once a day for 60 consecutive days.The contents of serum urea nitrogen and creatinine were determined by colorimetry.The contents of tumor necrosis factor-α(TNF-α),IL-6 and IL-1βof the kidney tissue,and the contents of TNF-α,IL-17 and matrix metalloproteinase-9(MMP-9)secreted by spleen lymphocytes were determined by ELISA.The activity of superoxide dismutase(SOD)and the level of malondialdehyde were determined by colorimetric method and colorimetric method,respectively.Pathological changes of rat kidney tissue were examined by HE staining.The protein levels of Toll-like receptor 4(TLR4)and NF-κB in rat kidney tissue were tested by Western blotting.Results:Compared with the model group,urea nitrogen and creatinine levels of the serum,TNF-α,IL-6,IL-1βand MDA levels,TLR4 and NF-κB protein levels of the kidney tissue,TNF-α,IL-17,MMP-9 levels of the spleen were significantly decreased in the low,medium and high dose groups of Tiaoqi Decoction(P<0.05),but SOD activity was significantly increased(P<0.05).The pathological changes of the kidney tissue was significantly attenuated.With the dose of Tiaoqi Decoction increasing,dose-dependent changes of the indicators above were observed.Conclusion:Self-made Tiaoqi Decoction could significantly attenuate renal injury induced by cisplatin in rats by inhibiting TLR4/NF-κB pathway.