加味二仙丸对支气管哮喘大鼠钙离子通道的影响

Effect of Jiawei Erxian Pill(加味二仙丸) on Calcium Ion Channel in Rats with Bronchial Asthma

目的:探讨加味二仙丸对支气管哮喘大鼠中钙库操控的钙离子通道(SOCC)的影响。方法:54只SD大鼠采用随机数字表法分成空白组9只,造模组45只。建立大鼠哮喘模型,造模成功后将造模组大鼠随机分成模型组、加味二仙丸低剂量组、加味二仙丸中剂量组、加味二仙丸高剂量组、固肾定喘丸组,每组6只。各药物组给予相应药物干预,模型组和空白组大鼠予以等容量的生理盐水进行灌胃,2次/d,连续给药28 d。通过HE染色判断哮喘模型是否制备成功,通过免疫组织化学法测基质相互作用分子1(STIM1)、钙释放激活钙通道蛋白1(ORAI1)的阳性表达,通过Western blotting法测STIM1、ORAI1蛋白相对表达量,通过荧光指示剂法观察钙离子平均荧光强度值,反映其钙离子浓度变化。结果:模型组大鼠支气管分支及其周围有大量以淋巴细胞为主的炎症细胞浸润,加味二仙丸低、中、高剂量组和固肾定喘丸组大鼠可见少量的炎症细胞浸润,各给药组大鼠损伤程度介于空白组和模型组之间,且各给药组损伤程度无明显差异。与空白组比较,模型组大鼠肺组织STIM1、ORAI1的平均光密度值和蛋白表达量、钙离子平均荧光强度值明显升高,加味二仙丸低剂量组大鼠肺组织STIM1的平均光密度值和蛋白表达量、ORAI1蛋白表达量、钙离子平均荧光强度值升高,加味二仙丸中剂量组和固肾定喘丸组大鼠肺组织ORAI1蛋白表达量升高,差异均有统计学意义(P<0.05)。与模型组比较,加味二仙丸高剂量组大鼠肺组织STIM1、ORAI1平均光密度值和蛋白表达量、钙离子平均荧光强度值明显降低,固肾定喘丸组大鼠肺组织STIM1和ORAI1蛋白表达量降低,加味二仙丸低、中剂量组大鼠肺组织ORAI1蛋白表达量降低,差异均有统计学意义(P<0.05)。结论:加味二仙丸能抑制哮喘大鼠肺组织中SOCC的激活,从而抑制钙离子内流,起到治疗支气管哮喘的作用。

Objective:To investigate the effect of Jiawei Erxian Pill on calcium ion channel(SOCC) controlled by calcium store in rats with bronchial asthma.Methods:54 SD rats were divided into blank group and modeling group by random number table method,with 9 in blank group and 45 in modeling group.The rat asthma model was established.After successful modeling,the rats in the modeling group was randomly divided into model group,low dose group of Jiawei Erxian Pill,medium dose group of Jiawei Erxian Pill,high dose group of Jiawei Erxian Pill and Gushen Dingchuan Pill(固肾定喘丸) group,with 6 rats in each group.Each drug group was given corresponding drug intervention.Rats in the model group and the blank group were given the same volume of normal saline by gavage,twice a day,for 28 consecutive days.He staining was used to determine whether the asthma model was successfully prepared.The positive expression of STIM1 and ORAI1 was measured by immunohistochemistry,the relative expression of STIM1 and ORAI1 was measured by Western blotting,and the average fluorescence intensity of calcium ions was observed by fluorescence indicator method to reflect the change of calcium ion concentration.Results:In the model group,there were a large number of inflammatory cells infiltrating the bronchial branches and their surroundings,mainly lymphocytes.A small number of inflammatory cells infiltrated in the low,medium and high dose groups of Jiawei Erxian Pill and the Gushen Dingchuan Pill groups.The injury degree of rats in each treatment group was between the blank group and the model group,and there was no significant difference in the injury degree of each treatment group.Compared with the blank group,the average optical density,protein expression and calcium ion average fluorescence intensity of STIM1and ORAI1 in the lung tissue of rats in the model group were significantly increased,the average optical density,protein expression,ORAI1 protein expression and calcium ion average fluorescence intensity of STIM1 in the lung tissue of rats in the low-dose group of Jiawei Erxian Pill were increased.The difference was statistically significant(P<0.05).Compared with the model group,the average optical density,protein expression and the average fluorescence intensity of calcium ions of STIM1 and ORAI1 in the lung tissue of rats in the high-dose group of Jiawei Erxian Pill were significantly decreased,the expression of STIM1 and ORAI1 in the lung tissue of rats in the Gushen Dingchuan Pill group was decreased,and the expression of ORAI1 protein in the lung tissue of rats in the low and medium dose groups of Jiawei Erxian Pill was decreased,with statistical significance(P<0.05).Conclusion:Jiawei Erxian Pill can inhibit the activation of SOCC in the lung tissue of asthmatic rats,and thus inhibit the influx of calcium ions,which plays a role in the treatment of bronchial asthma.