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目的:通过研究加味附子理中免煎颗粒对急性马兜铃酸肾病(AAN)大鼠肾小管上皮细胞凋亡的影响探讨其治疗急性AAN的作用机制。方法:将90只SD大鼠随机分为正常组、模型组、中药组、强的松组、中药+强的松组,其中正常组10只,其余各组均为20只。模型组、中药组、强的松组及中药+强的松组按100 mg.kg-1.d-1剂量予马兜铃酸A纯品连续ig 3 d建立AAN模型;从实验第6天开始,中药组予加味附子理中免煎颗粒(按生药量计为3.55 g.kg-1.d-1,10 mL.kg-1.d-1)ig,强的松组予强的松(0.5 mg.kg-1.d-1)ig,中药+强的松组予加味附子理中免煎颗粒(3.55 g.kg-1.d-1)+强的松(0.5 mg.kg-1.d-1)ig,模型组和正常组则予蒸馏水10 mL.kg-1.d-1 ig;连续给药3周后处死各组大鼠。以RT-PCR检测各组大鼠肾组织Bcl-2mRNA,Bax mRNA的表达,免疫组化法及Western blot法检测各组大鼠肾组织Bcl-2,Bax蛋白的表达,原位末端标记法(TUNEL)检测肾小管上皮细胞凋亡。结果:模型组大鼠肾组织Bcl-2 mRNA和蛋白表达明显减少,而Bax mRNA和蛋白表达增多;与模型组比较,中药组、强的松组和中药+强的松组均能不同程度增加Bcl-2 mRNA和蛋白的表达,降低Bax mRNA和蛋白的表达,差异均有统计学意义(P<0.05);其中以中药+强的松组的作用最佳(P<0.05)。TUNEL结果显示,正常组大鼠肾小管可见少量凋亡细胞,模型组则有大量凋亡的肾小管上皮细胞,差异有统计学意义(P<0.05);与模型组相比,中药组、强的松组和中药+强的松组凋亡细胞数均减少(P<0.05);而中药+强的松组凋亡细胞数减少最为明显(P<0.05)。结论:肾小管上皮细胞凋亡参与了急性马兜铃酸肾病的发病,加味附子理中免煎颗粒能减轻急性马兜铃酸肾病大鼠肾小管上皮细胞的凋亡,但联合强的松治疗疗效更佳。
Objective: To investigate the effect of Jiawei Fuzi Lizhong decoction on renal tubular epithelial cell apoptosis in acute aristolochic acid nephropathy (AAN) rats and its mechanism of action in treating acute AAN. Methods: Ninety SD rats were randomly divided into normal group, model group, traditional Chinese medicine group, prednisone group and traditional Chinese medicine + prednisone group, of which 10 in normal group and 20 in other groups. The model group, the traditional Chinese medicine group, the prednisone group and the traditional Chinese medicine + prednisone group were given AAN model at a dose of 100 mg.kg-1.d-1 for 3 days on a continuous ig for 3 days. From the 6th day At the beginning, the traditional Chinese medicine group was treated with flavonoids (3.55 g.kg-1.d-1, 10 mL.kg-1.d-1) (0.5 mg.kg-1.d-1) ig and traditional Chinese medicine + prednisone group were treated with Jiawei Fuzi Lizhong decoction granules (3.55 g.kg-1.d-1) + prednisone 1.d-1) ig, the model group and the normal group were given distilled water 10 mL.kg-1.d-1 ig; 3 weeks after continuous administration of rats were sacrificed. The expressions of Bcl-2mRNA and Bax mRNA were detected by RT-PCR. The expressions of Bcl-2 and Bax protein were detected by immunohistochemistry and Western blot. TUNEL) was used to detect renal tubular epithelial cell apoptosis. Results: Compared with model group, the expression of Bcl-2 mRNA and protein in renal tissue of rats in model group decreased significantly, while the expression of Bax mRNA and protein increased in model group. Compared with model group, Bcl-2 mRNA and protein expression, decreased Bax mRNA and protein expression, the difference was statistically significant (P <0.05), of which the traditional Chinese medicine + prednisone group the best (P <0.05). TUNEL results showed that a small amount of apoptotic cells were found in the renal tubules of normal rats and a large number of apoptotic renal tubular epithelial cells were found in the untreated group (P <0.05). Compared with the model group, (P <0.05), while the number of apoptotic cells in the Chinese medicine + prednisone group was the most significant (P <0.05). Conclusion: Apoptosis of renal tubular epithelial cells is involved in the pathogenesis of acute aristolochic acid nephropathy. Jiawei Fuzi Li Zhong Fei Jian Granule can reduce renal tubular epithelial cell apoptosis in acute aristolochic acid nephropathy rats, but combined with prednisone treatment Better effect.