三黄一龙汤对佐剂性关节炎继发心血管病变大鼠血管紧张素Ⅱ、醛固酮及相关炎性因子的影响

Effect of Sanhuang Yilong Formula on AngiotensinⅡ,Aldosterone and Related Inflammatory Factors in Rats with Cardiovascular Disease Secondary to Adjuvant Arthritis

目的:探讨三黄一龙汤对佐剂性关节炎大鼠心脏的病理组织学、肾素-血管紧张素-醛固酮系统(RAAS)效应因子血管紧张素Ⅱ(AngⅡ)、醛固酮(ALD)和炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达的影响,探讨三黄一龙汤对佐剂性关节炎大鼠继发心血管病变的干预作用及调节机制。方法:70只Wistar健康雌性大鼠按照随机数字表法分为正常组、模型组、阳性药物组、三黄一龙汤低剂量组、三黄一龙汤中剂量组、三黄一龙汤高剂量组和安慰剂组,每组10只。除正常组,其余各组大鼠建立佐剂性关节炎模型。三黄一龙汤低剂量组、三黄一龙汤中剂量组、三黄一龙汤高剂量组分别给予每日生药量19.8、39.6、79.2 g/kg灌胃,阳性药物组给予来氟米特2.1 mg/kg灌胃,安慰剂组灌胃等量生理盐水,连续干预4周。苏木精-伊红(HE)染色观察心脏的病理组织学变化,酶联免疫吸附测定法、蛋白印迹法和免疫组化法检测外周血和心脏组织AngⅡ、ALD、TNF-α和IL-6的变化。结果:病理结果显示,相对于正常组,模型组继发心肌损伤,主要表现为心肌组织发生局部炎症,局部心肌细胞排列紊乱,个别大鼠心肌有轻微间质水肿。方药三黄一龙汤能够显著改善心肌损伤,使其炎症病灶缩小,水肿程度减轻,下调心脏组织AngⅡ和ALD的表达(P<0.05),降低外周血和心脏组织TNF-α、IL-6水平(P<0.05)。结论:方药三黄一龙汤可能通过抑制RAAS中AngⅡ和ALD的表达,减少TNF-α、IL-6的产生,从而缓解佐剂性关节炎继发的心血管病变。

Objective:To investigate the effects of Sanhuang Yilong Formula on heart tissue,renin-angiotensin-aldosterone system(RAAS)and aldosterone(ALD)and inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)in the heart of rats with adjuvant arthritis.To explore the intervention effect on secondary cardiovascular disease in adjuvant arthritis rats.Methods:A total of 70 Wistar healthy female rats were randomly divided into normal group,model group,positive drug group,Sanhuang Yilong Formula low-dose group,Sanhuang Yilong Formula medium-dose group,Sanhuang Yilong Formula high-dose group and placebo group,with 10 rats in each group.In addition to the normal group,the other groups of rats constructed adjuvant arthritis model.Sanhuang Yilong Formula low-dose group,Sanhuang Yilong Formula medium-dose group,Sanhuang Yilong Formula high-dose group were given daily dose of 19.8,39.6 and 79.2 g/kg by gavage,respectively;positive drug group was given leflunomide 2.1 mg/kg by gavage;placebo group was given the equal volume of normal saline by gavage,and the intervention lasted for 4 weeks.Hematoxylin-eosin(HE)was used to observe the histopathological changes of the heart.The changes of AngⅡ,ALD,TNF-α,and IL-6 in peripheral blood and heart tissue were detected by enzyme-linked immunosorbent assay,Western Blot and immunohistochemistry.Results:The pathological results showed that compared with the normal group,the model group showed secondary myocardial injury,which was mainly manifested as local inflammation of myocardial tissue,disordered arrangement of local cardiomyocytes,and slight interstitial edema of individual myocardium.Sanhuang Yilong Formula could significantly improve myocardial injury,attenuate inflammatory lesions,reduce edema degree,down-regulated the expression of AngⅡand ALD in heart tissue(P<0.05),and decreased the levels of TNF-αand IL-6 in peripheral blood and heart tissue(P<0.05).Conclusion:Sanhuang Yilong Formula may alleviate secondary cardiovascular diseases after adjuvant arthritis by inhibiting the expression of Ang-Ⅱand ALD in the RAAS and attenuating IL-6 and TNF-αproduction.