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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 explore the mechanism of ephedrine tannin and ephedra on the treatment of asthma based on calcium channels. Methods: Sixty rats were randomly divided into blank group, control group, model group, ephedrine group, ephedrine tannic acid group, Maxing cough tablet low dose group, Maxing cough tablet high dose group. After constructing the asthma model, the number of inflammatory cells per unit area of the bronchial cross section and WAm / Pbm, WAi / Pbm were calculated by HE staining to evaluate airway inflammation. The expressions of CACC and T-MEM16A in rats were detected by immunohistochemistry and Western-blot. Results: (1) The number of inflammatory cells in the model group was higher than that in the blank group (P <0.01). Compared with the model group, the number of inflammatory cells, tannic acid group, ephedra group, Maxing cough tablet low dose group, Maxing cough tablet high dose group decreased (all P <0.01). WAm / Pbm, WAi / Pbm and tannic acid group decreased (P <0.01). There was no difference between the high dose group of Ephedra, the Maxing cough tablet low dose group and the Maxing cough tablet group compared with the model group. (2) The number of CACC1 positive cells in model group and blank control group was higher than that in control group, and the intensity of CACC1 was also higher than that of control group (P <0.01). Compared with model group, the ratio of optical density was calculated. The ratio of high dose group of tannin group, ephedra group, Maxing cough tablet low dose group and Maxing cough tablet group was decreased (P <0.01). CACC4 in the model group and the blank control group, the number of positive cells in the model group more than the control group, the intensity is also higher than the control group (P <0.01). Compared with the model group, the ratio of optical density was calculated. The ratio of high dose group of tannin group, ephedra group, Maxing cough tablet low dose group and Maxing cough tablet group was decreased (all P <0.01). (3) The expression of TMEM16A in the lung tissue of each group showed that the expression of TMEM16A in the lung tissue of the other groups were significantly decreased compared with the blank control group (P <0.01). Compared with the model control group, the expression levels of TMEM16A in the lung tissue of rats in each treatment group were significantly increased (P <0.05 or 0.01), of which TMEM16A The most significant increase in expression levels. Conclusions: (1) Ephedra tannin can decrease WAm / Pbm, WAi / Pbm and change airway remodeling. (2) The expression of CACC1 and CACC4 in lung tissue of both low-dose and low-dose groups of Tannin, Ephedra, Maxing Cough-Relieving Tablet and Maxing Cough-Treating Tablet can down-regulate the expression of TMEM16A.