祛风解痉方调节γδT细胞抑制哮喘小鼠气道高反应性的分子机制(1)
[摘要] 目的 探討祛风解痉方调节γδT细胞抑制哮喘小鼠气道高反应性的分子机制。 方法 以卵清蛋白滴鼻的方法建立哮喘Balb/c小鼠模型，以祛风解痉方药进行干预，比较祛风解痉方对γδT细胞活化过程中信号通路分子磷酸化水平的干预效果。30只实验小鼠按照随机数字表法分为6组，每组5只，分别为空白对照组、哮喘模型组、醋酸泼尼松组(0.27 mg/d)、祛风解痉方低剂量组(0.59 g/d)、祛风解痉方中剂量组(1.18 g/d)、祛风解痉方高剂量组(2.36 g/d)。各组分别进行干预，1次/d，共给药7 d。给药结束后麻醉处死小鼠，分离纯化及扩增肺气道组织γδT细胞，CCK-8检测γδT细胞的增殖活性，Western blot检测不同信号通路分子的磷酸化水平。 结果 与空白对照组比较，哮喘模型组γδT细胞活性明显增强，差异有统计学意义(P < 0.05)。与哮喘模型组比较，祛风解痉方低剂量组的γδT细胞活性差异无统计学意义(P > 0.05)，而祛风解痉方中、高剂量组以及醋酸泼尼松组的γδT细胞活性明显降低，差异有统计学意义(P < 0.05)。与空白对照组比较，哮喘模型组p-ERK1/2、p-P38、β-catenin的蛋白表达量增加，差异有统计学意义(P < 0.05)，ERK1/2、P38蛋白表达水平差异无统计学意义(P > 0.05)。与哮喘模型组比较，祛风解痉方低、中、高剂量组以及醋酸泼尼松组p-ERK1/2、p-P38、β-catenin的蛋白表达量减少，差异有统计学意义(P < 0.05)，而祛风解痉方中、高剂量组以及醋酸泼尼松组ERK1/2蛋白表达水平增加，祛风解痉方低剂量组以及醋酸泼尼松组P38蛋白表达水平增加，差异有统计学意义(P < 0.05)。 结论 祛风解痉方调节γδT细胞抑制哮喘小鼠气道高反应性的可能分子信号通路机制为抑制p-ERK1/2、p-P38、β-catenin的蛋白表达量，减轻哮喘小鼠气道高反应性，达到治疗哮喘的目的。
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[关键词] 祛风解痉方;哮喘;γδT淋巴细胞;信号通路分子
[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-7210(2020)02(c)-0004-05
[Abstract] Objective To investigate the molecular mechanism of Qufeng Jiejing Prescription in regulating γδT cells to inhibit airway hyperresponsiveness in asthmatic mice. Methods The Balb/c mouse model of asthma was established with the method of ovalbumin nasal drip， and the intervention was conducted with the drugs of Qufeng Jiejing Prescription. The intervention effect of Qufeng Jiejing Prescription on the molecular phosphorylation level of signaling pathway during activation of γδT cells was compared. Thirty experimental mice method was divided into 6 groups according to random number table， each group of 5 mice， respectively for the blank control group， asthma model group， Prednisone Acetate group (0.27 mg/d)， Qufeng Jiejing Prescription low dose group (0.59 g/d)， Qufeng Jiejing Prescription medium dose group (1.18 g/d)， Qufeng Jiejing Prescription high dose group (2.36 g/d). Each group was treated 1 time/d and intervention for 7 days. After administration， the mice were anesthetized and sacrificed， and the γδT cells in the lung airway tissues were isolated， purified and amplified. The proliferation activity of the γδT cells was detected by CCK-8， and the phosphorylation levels of molecules in different signaling pathways were detected by Western blot. Results Compared with the blank control group， the activity of γδT cells in the asthma model group was significantly increased， with statistically significant difference (P < 0.05). Compared with the asthma model group， there was no significant difference in the activity of γδT cells in the Qufeng Jiejing Prescription low dose group (P > 0.05)， while the activity of γδT cells in the medium， high dose groups of Qufeng Jiejing Prescription and the Prednisone Acetate group were significantly decreased (P < 0.05). Compared with the blank control group， the protein expression levels of p-ERK1/2， p-P38 and β-catenin in the asthma model group increased， with statistically significant difference (P < 0.05)， while the protein expression levels of ERK1/2 and P38 showed no statistically significant difference (P > 0.05). Compared with the asthma model group， the protein expression levels of p-ERK1/2， p-P38 and β-catenin in the low， medium and high dose groups of Qufeng Jiejing Prescription and the Prednisone Acetate group were decreased， with statistically significant differences (P < 0.05). However， ERK1/2 protein expression level increased in the middle and high dose groups of Qufeng Jiejing Prescription and Prednisone Acetate group， and P38 protein expression level increased in the Qufeng Jiejing Prescription low dose group and Prednisone Acetate group， the differences were statistically significant (all P < 0.05). Conclusion The possible molecular signaling pathway mechanism of the regulation of T cells to inhibit airway hyperresponsiveness in asthmatic mice is to inhibit the protein expression levels of p-ERK1/2， p-P38 and β-catenin， so as to reduce airway hyperresponsiveness in asthmatic mice and achieve the treatment of asthma., 百拇医药(樊长征 苗青 洪巧瑜)
, http://www.100md.com
[关键词] 祛风解痉方;哮喘;γδT淋巴细胞;信号通路分子
[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-7210(2020)02(c)-0004-05
[Abstract] Objective To investigate the molecular mechanism of Qufeng Jiejing Prescription in regulating γδT cells to inhibit airway hyperresponsiveness in asthmatic mice. Methods The Balb/c mouse model of asthma was established with the method of ovalbumin nasal drip， and the intervention was conducted with the drugs of Qufeng Jiejing Prescription. The intervention effect of Qufeng Jiejing Prescription on the molecular phosphorylation level of signaling pathway during activation of γδT cells was compared. Thirty experimental mice method was divided into 6 groups according to random number table， each group of 5 mice， respectively for the blank control group， asthma model group， Prednisone Acetate group (0.27 mg/d)， Qufeng Jiejing Prescription low dose group (0.59 g/d)， Qufeng Jiejing Prescription medium dose group (1.18 g/d)， Qufeng Jiejing Prescription high dose group (2.36 g/d). Each group was treated 1 time/d and intervention for 7 days. After administration， the mice were anesthetized and sacrificed， and the γδT cells in the lung airway tissues were isolated， purified and amplified. The proliferation activity of the γδT cells was detected by CCK-8， and the phosphorylation levels of molecules in different signaling pathways were detected by Western blot. Results Compared with the blank control group， the activity of γδT cells in the asthma model group was significantly increased， with statistically significant difference (P < 0.05). Compared with the asthma model group， there was no significant difference in the activity of γδT cells in the Qufeng Jiejing Prescription low dose group (P > 0.05)， while the activity of γδT cells in the medium， high dose groups of Qufeng Jiejing Prescription and the Prednisone Acetate group were significantly decreased (P < 0.05). Compared with the blank control group， the protein expression levels of p-ERK1/2， p-P38 and β-catenin in the asthma model group increased， with statistically significant difference (P < 0.05)， while the protein expression levels of ERK1/2 and P38 showed no statistically significant difference (P > 0.05). Compared with the asthma model group， the protein expression levels of p-ERK1/2， p-P38 and β-catenin in the low， medium and high dose groups of Qufeng Jiejing Prescription and the Prednisone Acetate group were decreased， with statistically significant differences (P < 0.05). However， ERK1/2 protein expression level increased in the middle and high dose groups of Qufeng Jiejing Prescription and Prednisone Acetate group， and P38 protein expression level increased in the Qufeng Jiejing Prescription low dose group and Prednisone Acetate group， the differences were statistically significant (all P < 0.05). Conclusion The possible molecular signaling pathway mechanism of the regulation of T cells to inhibit airway hyperresponsiveness in asthmatic mice is to inhibit the protein expression levels of p-ERK1/2， p-P38 and β-catenin， so as to reduce airway hyperresponsiveness in asthmatic mice and achieve the treatment of asthma., 百拇医药(樊长征 苗青 洪巧瑜)