慢性马兜铃酸肾病大鼠肾组织微血管损伤机制及温阳活血方的干预作用

Injury Mechanism of Renal Microvessel and Intervention Effect of Wenyanghuo xuefang on Rats with Chronic Aristolochic Acid Nephropathy

目的:研究慢性马兜铃酸肾病大鼠肾组织微血管损伤机制及温阳活血方对其干预作用,探讨温阳活血方对慢性马兜铃酸肾病的保护作用机制。方法:将48只雄性SD大鼠随机分为5组。(1)正常对照组(n=8):予生理盐水灌胃;(2)模型组(n=10):按关木通水煎液10ml·kg-1·d-1(相当于关木通40g·kg-1·d-1,马兜铃酸A2.6mg·kg-1·d-1)给大鼠灌胃;(3)中药组(n=10):在模型组基础上,再予温阳活血方30g·kg-1·d-1灌胃;(4)西药组(n=10):在模型组基础上,再予科素亚33.3mg·kg-1·d-1灌胃;(5)中西药结合组(n=10):在模型组基础上,再予温阳活血方30g·kg-1·d-1+科素亚33.3mg·kg-1·d-1灌胃。20周末,光镜观察肾脏病理,采用CD34免疫组化染色来反映肾组织微血管的损伤情况,实时PCR检测肾组织中Ang-1、Ang-2、Tie-2和VEGF mRNA的表达。结果:(1)与正常对照组比较,模型组大鼠肾组织CD34表达明显降低(P<0.01);而治疗组大鼠肾组织CD34表达较模型组明显升高(P<0.05),其中尤以中药组明显(P<0.01);(2)与正常对照组比较,模型组大鼠肾组织Ang-1、Tie-2、VEGF mRNA表达明显降低(P<0.05,P<0.01),Ang-2表达明显升高(P<0.01);而治疗组大鼠肾组织Ang-1、Tie-2、VEGFmRNA表达较模型组明显升高(P<0.05,P<0.01),中西药结合组Ang-2表达明显减少(P<0.01)。结论:慢性马兜铃酸肾病大鼠肾组织存在微血管的损伤,其损伤机制可能与Ang-1、Tie-2和VEGF mRNA的下调,Ang-2 mRNA上调有关,温阳活血方能明显改善微血管的损伤,对慢性马兜铃酸肾病大鼠肾脏具有保护作用。

Objective：To explore the renal protective mechanism of Wen Yang Huo Xue Fang （WYHXF） by investigating the injury mechanism of renal microvessel in rats with chronic aristolochic acid nephropathy. Methods： Fourty- eight male SD rats were randomly divided into five groups. Group A （ n = 8 ） were treated with normal saline （ 10 ml·kg^- 1· d^- 1 ）. Group B（ n = 10 ） were treated with caulis Aristolochia Manshuriensis decoction （Aristolochia Manshuriensis 40 g·kg^-1· d^- 1, Aristolochic acid A 2.6 mg·kg^- 1· d^-1 ）. Group C （ n = 10） were treated with WYHXF （ 30 g·kg^- 1· d^-1 ） after the mice being modeled with the above -mentioned method. Group D （n = 10） were treated with Losartan Potassium Tablets （33.3 mg·kg^- 1· d^-1） after the mice being modeled and the above - mentioned method. Group E（ n = 10） were treated with WYHXF （30 g·kg^- 1· d^-1） and Losartan Potassi- um Tablets （33.3 mg·kg^- 1· d^-1 ） after the mice being modeled and the above - mentioned method. After 20 weeks, observation of renal pathology, renal microvessel injury was indirectly assessed by CD34 irnmunohistochemistry. The mRNA expression of Ang- 1, Ang- 2,Tie- 2and VEGF were detected by realtime PCR. Results： （1） Compared with that in the normal control group, the expression of CD34 was significantly decreased in the model group （ P 〈 0.01 ） ; Compared with that in the model group, the expression of CI）34 was significantly elevated in the therapy group （P〈0.05, P〈0.01）. （2） Compared with those in the normal control group, the mRNA expressions of Ang - 1, Tie - 2 and VEGFmRNA were significantly decreased （ P 〈 0.05, P 〈 0.01 ）, while the mRNA expression of Ang- 2 was significantly elevated （P 〈 0, 01 ） in the model group. Compared with those in the model group, the mRNA expressions of Ang - 1, Tie- 2, VEGFmRNA were elevated （ P 〈 0.05, P 〈 0.01 ）, while the mRNA expression of Ang - 2 was decreased in the therapy group（ P 〈 0,01 ）. Conclusion： There is renal microvessel injury in AAN. The mechanism may be related to reduction of Ang - 2 and elevation of Ang - 1, Tie - 2 and VEGF. WYHXF can significantly relieve microvascular injury, which exerted protective effect on CAAN.