射麻止喘汤对哮喘大鼠支气管平滑肌细胞增殖的影响_临床医学论文.docVIP

射麻止喘汤对哮喘大鼠支气管平滑肌细胞增殖的影响_临床医学论文 射麻止喘汤对哮喘大鼠支气管平滑肌细胞增殖的影响_临床医学论文 作者：刘建博，张玉芳，周宇，黄桂南，郭敏 【摘要】 　　【目的】观察射麻止喘汤对支气管哮喘大鼠气道平滑肌细胞（ASMC）增殖的影响。【方法】选用SPF级SD大鼠分为哮喘组和正常对照组，哮喘组采用卵清蛋白（OVA）致敏并激发方法复制哮喘模型，造模成功后分别取各组支气管平滑肌进行细胞培养，倒置显微镜观察形态并经免疫荧光染色鉴定；给予大鼠灌胃高、中、低剂量的射麻止喘汤（剂量分别为64、32、6.4g·kg-1·d-1）制备含药血清；取2~5代ASMC分别给予射麻止喘汤高、中、低剂量含药血清，蛋白激酶C（PKC）激活剂佛波醇肉豆蔻酰乙酸佛波酯（PMA，10nmol/L），PKC抑制剂马来酰亚胺甲磺酸盐（Ro，5μmol/L）干预；另设空白组（不给予任何药物血清干预）。采用四甲基偶氮唑盐（MTT）法检测各组ASMC增殖（D值）。【结果】免疫荧光染色鉴定结果显示所培养的细胞为平滑肌细胞。正常大鼠PMA组ASMC的D值较空白组显著增高（P＜0.05），Ro组、射麻止喘汤各剂量组D值与空白组比较差异均无显著性意义（P＞0.05）。哮喘模型大鼠PMA组ASMC的D值较空白组显著增加（P＜0.05），Ro组、射麻止喘汤各剂量组D值均较空白组显著降低（P＜0.05），但射麻止喘汤各剂量组与Ro组比较差异无显著性意义（P＞0.05）。【结论】射麻止喘汤治疗支气管哮喘的作用可能与其能抑制ASMC增殖，影响哮喘的气道重塑过程有关。 【关键词】 射麻止喘汤/药理学；支气管哮喘/中药疗法；平滑肌细胞/病理学；细胞增殖；细胞培养；疾病模型，动物；大鼠 　　Abstract: ObjectiveTo observe the effect of Shema Zhichuan Decoction（SZD） on the proliferation of the airway smooth muscle cells（ASMC） in asthmatic rats. MethodsSpecificsensitized with intraperitoneal injection of ovalbumin（OVA） and challenged with intranasal application of OVA. After successful modeling，bronchial smooth muscle cells were sampled for culturing，and their histological features were observed under invert microscope. Immunofluenscent assay was used for identification. Serum containing SZD was prepared after administration of SZD in the dosages of 64，32 and 6.4 g·kg-1·d-1 to the rats by gastric gavage. The subculture ASMC of the 2nd to 5th generation were treated high，moderate，protein kinase C（PKC） activator of phorbol myristate acetate（PMA，10 nmol/L） and PKC inhibitor of maleimide methanesulfonate（RO，5 μmol/L ），respectively. A blank control group was also set up. The proliferation of ASMC was detected by methyl thiazolyl tetrazolium（MTT） assay. ResultsThe cultured cells were identified as bronchial smooth muscle cells by immunofluenscent assay. For the normal rats，the absorption value of ASMC was higher in PMA group than that in the blank control group（0.05），but absorption value in RO（Pgt;0.05）. As for the asthmatic rats，the absorp