加减射麻止喘汤对哮喘大鼠支气管平滑肌细胞生长周期的干预作用

Intervention effects of the addition and subtraction of Shema Zhichuan decoction on the growth cycle of bronchial smooth muscle cells of asthma rat

目的探讨加减射麻止喘汤对哮喘大鼠支气管平滑肌细胞(ASMC)生长周期的干预作用。方法以卵清蛋白(OVA)致敏并激发制造大鼠哮喘模型,提取ASMC进行细胞传代培养;高、中、低剂量的加减射麻止喘汤分别灌胃大鼠以制备含药血清,以含药血清、蛋白激酶C(PKC)激活剂佛波醇-12-肉豆蔻酰-13-乙酸佛波酯(PMA),PKC抑制剂马来酰亚胺甲磺酸盐(Ro-31-8220)分别干预3～5代ASMC,空白组不予药物干预。采用四甲基偶氮唑盐(MTT)法检测各组ASMC增加、流式细胞术检测ASMC各周期分布。结果与空白组、PMA组相比,加减射麻止喘汤高、中、低剂量组的D值显著降低,加减射麻止喘汤高剂量组的G1期、G2/M期细胞百分比显著升高,S期细胞百分比显著降低(P<0.05);与Ro-31-8220组相比,加减射麻止喘汤高低剂量组的D值显著增加,加减射麻止喘汤高剂量组的G1期细胞百分比显著降低,S期细胞百分比显著升高(P<0.05)。结论加减射麻止喘汤可抑制哮喘大鼠ASMC的细胞增殖,从而影响气道重构。

Objective To explore the intervention effects of the addition and subtraction of Shema Zhichuan decoction on the growth cycle of bronchial smooth muscle cells of asthma rat （ASMC）. Methods The asthma rat model was sensitized and challenged by ovalbumin （OVA）. Then, ASMC cells were isolated and cultured. The rat serum containing different contents of addition and subtraction of Shema zhichuan decoction were prepared by gavaged with high dose, medium dose and low dose of medicines, respectively. ASMCs from 3th to 5th generations were treated respec- tively with serum contained with traditional Chinese medicines, protein kinase C （PKC） activating a- gent phorbol 12 myristate 13 acetate （PMA）, PKC inhibitor maleic imide methyl sulfonate （Ro-31- 8220）. The ASMCs proliferation was determined using MTT assay （D value）, the cell cycle division percentage of ASMCs was detected using flow cytometry. Results Compared with blank group and PMA group, the ASMCs proliferations in high dose group, medium dose group and low dose group of addition and subtraction of Shema Zhichuan decoction were significantly decreased, and the centages of ASMCs in G1 phase and G2/M phase were increased and the percentage of ASMCs per- in Sphase was decreased significantly（P〈0.05） in high dose group of addition and subtraction of She- ma zhichuan decoction. Compared with Ro-31-8220 group, the ASMCs proliferations were in- creased significantly in Shema Zhichuan decoction high dose group and low dose group, the per- centage of ASMCs in Gl phase was decreased and that of in S phase was increased significantly in high dose group of addition and subtraction of Shema Zhichuan decoction（P〈0.05）. Conclusion The addition and subtraction of Shema Zhichuan decoction can inhibit the proliferation of asthma rat ASMC＇s cell, and then affect the bronchial wall remodeling.