基于钙离子通道探讨麻黄鞣酸及麻黄等治疗哮喘作用机制研究

Study on the Mechanism of Ephedra Tannic Acid and Ephedra in the Treatment of Asthma Based on Calcium Channel

目的 :基于钙离子通道探讨麻黄鞣酸及麻黄等治疗哮喘作用机制研究。方法 :60只大鼠随机分为空白组、对照组、模型组、麻黄组、麻黄鞣酸组、麻杏止咳片低剂量组、麻杏止咳片高剂量组。构建哮喘模型后,采用HE染色计算哮喘大鼠支气管横断面单位面积的炎性细胞数以及WAm/Pbm,WAi/Pbm,以评估气道炎症。用免疫组化法及蛋白质免疫印迹法(Western-blot)测定大鼠CACC和T MEMl6A蛋白表达。结果 :(1)炎性细胞数,模型组高于空白组(P<0.01)。与模型组比较,炎性细胞数,鞣酸组、麻黄组、麻杏止咳片低剂量组、麻杏止咳片高剂量组降低(均P<0.01)。WAm/Pbm,WAi/Pbm,鞣酸组下降(P<0.01),麻黄组、麻杏止咳片低剂量组、麻杏止咳片组高剂量组与模型组比较无差异。(2)CACC1在模型组和空白对照组,模型组阳性表达细胞数目多于对照组,强度也高于对照组(P<0.01)。与模型组比,计算其光密度比值,鞣酸组、麻黄组、麻杏止咳片低剂量组、麻杏止咳片组高剂量组比值下调(P<0.01)。CACC4在模型组和空白对照组,模型组阳性表达细胞数目多于对照组,强度也高于对照组(P<0.01)。与模型组比较,计算光密度比值,鞣酸组、麻黄组、麻杏止咳片低剂量组、麻杏止咳片组高剂量组比值下调(均P<0.01)。(3)各组大鼠肺组织中TMEM16A的表达检测结果显示:与空白对照组比较,其余各组大鼠肺组织中TMEM16A的表达水平均显著下调,差异有统计学意义(P<0.01);与模型对照组比较,各治疗组大鼠肺组织中TMEM16A的表达水平均显著上调,差异有统计学意义(P<0.05或0.01);其中麻杏止咳片高剂量组大鼠肺组织中TMEM16A的表达水平上调最为显著。结论 :(1)麻黄鞣酸可以使WAm/Pbm、WAi/Pbm下降,可以改变气道重构。(2)鞣酸组、麻黄组、麻杏止咳片低剂量组、麻杏止咳片高剂量组均可以下调CACC1、CACC4在肺组织中的表达,也都可以上调TMEM16A的表达。

Objective ：To study the mechanism of Ephedra tannic acid and Ephedra in the treatment of asthma based on calcium channel. Methods ：60 rats were randomly divided into blank control group,model group,tannic acid,ephedra,hemp apricot cough low dose group,hemp apricot cough high dose group. After asthma model was constructed,HE staining was used to calculate the unit cross-sectional area of the bronchial asthma rats the number of inflammatory cells and WAm/Pbm,WAi/Pbm,in order to assess airway inflammation. The expressions of CACC and TMEM16 A proteins in rats were determined by immunohistochemistry and Western blotting（Western-blot）. Results ：（1）The number of inflammatory cells,model group was higher than control group（P0.01）. Compared with the model group,the number of inflammatory cells,tannic acid group,ephedra group,hemp apricot cough low dose group,hemp apricot cough high dose group were decreased（P0.01）. WAm/Pbm,WAi/Pbm index,tannic acid group was decreased（P0.01）,compared with model group,ephedra group,low dose group,high dose group had no difference.（2）CACC1 in model group and blank control group,model group was higher than in the control group with positive cells,the intensity was higher than that of the control group（P0.01）. Compared with the model group,calculate the optical density ratio,tannic acid,Ephedra group,low dose group,high dose group reduced the ratio（P0.01）.CACC4 in model group and blank control group,model group than in the control group with positive cells,the intensity was higher than that of the control group（P0.01）. Compared with the model group,calculated the optical density ratio,tannic acid,Ephedra group,low dose group,high dose group reduced the ratio（P0.01,P0.01,P0.01,P0.01）.（3）Results showed that ：compared with the control group,the expression of TMEM16 A in lung tissue of rats in other groups were decreased,the difference was statistically significant（P0.01）;compared with the model group,the expression of TMEM16 A in lung tissue of rats in each treatment group were significantly increased,the difference was statistically significant（P0.05 or 0.01）;the high dose expression level increased most significantly. Conclusion ：（1）Ephedra tannic acid can make WAm/Pbm,WAi/Pbm decreased,can change the airway remodeling.（2）tannic acid group,Ephedra group,low dose group,high dose group can reduce CACC1,CACC4 expression in the lung tissue,also can increase the expression of TMEM16 A.