复方青秦液对尿酸性肾病大鼠肾组织血管紧张素Ⅱ及环氧化酶-2影响的研究

Effect of Compound Qingqin Liquid on the Expression Levels of AngⅡ and COX-2 mRNA Transcription and Protein Expression in the Renal Tissue of Uric Acid Nephropathy Rats:an Experimental Study

目的探讨复方青秦液对尿酸性肾病大鼠肾组织中血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)、环氧化酶-2(cyclooxygenase-2,COX-2)mRNA转录及蛋白表达水平的影响。方法将SD大鼠按照编号随机取码原则随机分为空白对照组、模型组、阳性对照组、中药大、中、小剂量组,用腺嘌呤灌胃伴饲酵母造模。空白对照组及模型组每日灌胃蒸馏水;阳性对照组每日灌胃别嘌醇9.33mg/kg;中药大、中、小剂量组每日给予复方青秦液3.77、1.89、0.09g/kg灌胃,连续6周。在4周时每组随机部分大鼠(空白组3只,其余每组6只),6周时处死剩余大鼠,取肾组织采用RT-PCR扩容AngⅡ和COX-2mRNA转录水平,采用ELISA测定AngⅡ蛋白表达水平,采用Western blot电泳技术及免疫组化测定COX-2蛋白表达水平。结果与空白对照组比较,模型组除4周AngⅡmRNA表达,4、6周时AngⅡ、COX-2mRNA及蛋白表达均明显升高(P<0.01,P<0.05)。4周时,中药大、中剂量组COX-2蛋白表达明显低于模型组及中药小剂量组(P<0.05),阳性对照组COX-2蛋白平均积分光密度值也明显低于模型组(P<0.01)。6周时,与模型组比较,除中药大剂量组AngⅡmRNA表达外,阳性对照组及中药各剂量组AngⅡmRNA及蛋白表达均降低(P<0.05,P<0.01),其中中药大、中剂量组效果优于阳性对照组及中药小剂量组(P<0.05,P<0.01);另外,阳性对照组及中药各剂量组COX-2mRNA表达、平均积分光密度值以及阳性对照组、中药大、中剂量组COX-2蛋白表达均明显低于模型组(P<0.05),其中中药中剂量组COX-2mRNA表达优于阳性对照组(P<0.05),中药大剂量组COX-2蛋白表达优于中药小剂量组(P<0.05)。结论复方青秦液可以下调尿酸性肾病大鼠肾脏AngⅡ、COX-2mRNA转录及蛋白表达水平,从而抑制肾组织炎性病理损伤。

Objective To investigate the effect of Compound Qingqin Liquid （CQL） on the ex- pression level of angiotensin II （Ang II ） and COXo2 mRNA transcription and protein expression in the re- nal tissue of rats with uric acid nephropathy. Methods SD rats were randomly divided into the blank con- trol group, the model group, the positive drug group, the high, moderate, and low dose CQL group according to number randomization principle. The model was established by gastrogavage of adenine, accompanied with yeast feeding. Distilled water was given by gastrogavage to rats in the blank control group and the model group. AIIopurinol at the daily dose of 9.33 mg/kg was given by gastrogavage to rats of the positive control group. CQL at the daily dose of 3.77 g/kg, 1.89 g/kg, and 0.09 g/kg was respectively given by gastrogavage to rats in the high, moderate, and low dose CQL groups. All treatment lasted for 6 weeks. Rats were randomly divided at week 4 （3 in the blank control group, and 6 in the rest groups）, and the rest rats were killed at week 6. The renal tissue was extracted. The expression level of Ang II and COX-2 mRNA transcription were detected by RT-PCR. The expression level of Ang II was detected by ELISA. The expression level of COX-2 protein was detected by Western blot and immunohistochemical assay. Results Compared with the blank control group, except the mRNA expression of Ang II at week 4, the mRNA and protein expression of Ang II and COX-2 obviously increased at week 4 and 6 in the model group （P 〈0.01, P 〈0.05）. The COX-2 protein expression at week 4 was obviously lower in the high and moderate dose CQL groups than in the model group and the low dose CQL group （P 〈0.05） ; the average integral of optical density value was obviously lower in the positive control group than in the model group. Except the mRNA expression of Ang II in the high dose CQL group at week 6, the mRNA and protein expression of Ang 1I obviously decreased in the positive control group and each dose CQL group （P 〈0.01, P 〈0.05）. Of them, the effects were better in the high and moderate dose CQL groups than in the positive control group and the low dose CQL group （P 〈0.05, P 〈0.01）. Besides, the mRNA expression of COX- 2, the average integral of optical density value were obviously lower in the positive control group and each dose CQL group than in the model group （P 〈0.05）. The protein expression of COX-2 was obviously lower in the high and moderate dose CQL groups than in the model group （P 〈0.05）. Of them, the mRNA expression of COX-2 was better in the moderate dose CQL group than in the positive control group （P 〈 0.05） ; the protein expression of COX-2 was better in the high dose CQL group than in the low dose CQL group （P 〈0.05）. Conclusion CQL was capable of lowering the expression level of Ang g, COX-2 mRNA transcription and protein expression, thus suppressing the inflammatory pathological injury of the renal tissue.