复方青秦液对尿酸性肾病大鼠肾组织Toll样受体表达的影响

Effect of Compound Qingqin Liquid on the Expression of Toll-like Receptor in the Renal Tissue of Rats with Urate Nephropathy

目的观察复方青秦液对尿酸性肾病大鼠肾组织Toll样受体TLR2、TLR4基因及蛋白表达的影响,探讨其肾脏保护的机制。方法将55只SD大鼠随机分为正常组5只;模型组、阳性药组及中药大、中、小剂量组,每组各10只,用腺嘌呤灌胃伴饲酵母造模。正常组及模型组每日灌胃蒸馏水10 mL/kg,阳性药组每日灌胃别嘌醇9.33 mg/kg,中药大、中、小剂量组每日灌胃复方青秦液3.77、1.89、0.94 g/kg,连续6周。4周和6周时分别处死部分大鼠,留取肾脏组织,RT-PCR检测TLR2、TLR4 mRNA转录水平,Western blot测定TLR2、TLR4蛋白表达水平,免疫组化测定TLR4蛋白表达水平。结果与正常组同期比较,4、6周模型组大鼠肾组织TLR4 mRNA转录水平、TLR2、TLR4蛋白表达水平升高(P<0.05,P<0.01)。与模型组同期比较,4、6周复方青秦液各剂量组TLR2、TLR4mRNA转录水平差异均无统计学意义(P>0.05),6周复方青秦液各剂量组TLR2、TLR4蛋白表达水平降低(P<0.05,P<0.01)。结论复方青秦液可通过降低TLR2、TLR4蛋白表达以抑制肾组织炎性病理损伤。

Objective To investigate the effect of Compound Qingqin Liquid （CQL） on Toll-like receptor 2 （TLR2） and Toll-like receptor 4 （3-LR4） in rats with urate nephropathy, and to explore its renal protection mechanism. Methods Totally 55 SD rats were randomly divided into 5 groups,i.e., the normal control group （n =5）, the model group （n =10）, the positive drug group （n =10）, and the high-, medi- um-, low-dose CQL groups （n =10） respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. AIIopurinol was intragas- trically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intra- gastrically administered at the daily dose of 3.77, 1.89,0.94 g/kg to rats in the high-, medium-, and low- dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively.Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction （ RT-PCR）. The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay. Results At week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group （P 〈 0.05, P 〈0.01）. Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4mRNA among the 3 CQL groups （P 〉0.05）at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL （P 〈0.05, P 〈0.01 ）. Conclusion CQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.