益气定眩饮通过Nod样受体蛋白-3炎性小体调节糖尿病合并缺血性脑卒中大鼠炎症因子的研究

Study of Yiqi Dingxuan Decoction(益气定眩饮)Regulating Inflammatory Factors in Diabetic Rats with Ischemic Stroke through Nod-like Receptor Protein-3 Inflammasome

目的:探讨益气定眩饮调节糖尿病合并缺血性脑卒中大鼠Nod样受体蛋白-3(NLRP3)炎性小体及白介素-18(IL-18)、IL-1β炎症因子的作用及机制。方法:将130只SD雄性大鼠复制糖尿病合并缺血性脑卒中动物模型并进行研究,成模后采用随机数字表法分为空白组、模型组、益气定眩饮组、丁苯酞胶囊组、益气定眩饮联合丁苯酞胶囊组。分别给予蒸馏水及相应药物干预3 d后,收集大鼠血清及脑组织。采用神经功能评分法评估大鼠神经功能变化;苏木精-伊红染色法观察脑组织形态结构;酶联免疫吸附测定检测血清炎症因子IL-18、IL-1β水平;逆转录PCR检测NLRP3炎症小体及半胱天冬酶-1(Caspase-1)表达。结果:与空白组比较,模型组均可改变大鼠脑组织结构,神经功能评分、IL-18水平、IL-1β水平、NLRP3 mRNA及Caspase-1 mRNA表达升高(P<0.05);与模型组比较,用药组均可改善大鼠脑病变组织结构,神经功能评分、IL-18水平、IL-1β水平、NLRP3 mRNA及Caspase-1 mRNA表达降低(P<0.05),且与单一用药比较,联合用药组IL-18水平、IL-1β水平、NLRP3 mRNA及Caspase-1 mRNA表达降低(P<0.05)。结论:益气定眩饮可改善糖尿病合并缺血性脑卒中大鼠脑损害,降低脑NLRP3、Caspase-1蛋白及血清IL-18、IL-1β表达,减轻炎症状态,发挥治疗作用,且联合丁苯酞用药效果更佳。

Objective:To investigate the effect and its mechanism of Yiqi Dingxuan Decoction on NLRP3 inflammasome and IL-18,IL-1βinflammatory factors in diabetic rats with ischemic stroke.Methods:130 male SD rats were used to establish the animal model of diabetes combined with ischemia stroke and conduct researth,the rats were randomly divided into blank group,model group,Yiqi Dingxuan Decoction group,butylphthalide capsule group,Yiqi Dingxuan Decoction combined with butylphthalide capsule group.The rats were treated with distilled water and corresponding drugs for 3 days,then the serum and brain tissues were collected.The changes of neural function were evaluated by neurological function scoring,the brain tissue was observed by HE staining,the levels of serum inflammatory factors IL-18 and IL-1βwere measured by enzyme-linked immunosorbent assay,and the expression of NLRP3 inflammasome and Caspase-1 was detected by RT-PCR.Results:Compared with the blank group,the brain structure of the model group was changed,and the neural function score,the content of IL-18 and IL-1β,the expression of NLRP3 mRNA and Caspase-1 mRNA were increased(P<0.05).Compared with the model group,the expression of NLRP3 mRNA and Caspase-1 mRNA,the content of IL-18 and IL-1β,the expression of NLRP3 mRNA and Caspase-1 mRNA in drug groups were decreased(P<0.05),and compared with the single drug group,the content of IL-18 and IL-1β,the expression of NLRP3 mRNA and Caspase-1 mRNA in the combined drug group decreased(P<0.05).Conclusion:Yiqi Dingxuan Decoction can improve the brain damage of diabetic rats with ischemic srtoke,decrease the expression of NLRP3,Caspase-1 protein and serum IL-18,IL-1β,relieve the inflammatory state,and play a therapeutic role,and the combination of butylphthalide has better effect.