目的 研究益气除痰方对缺氧条件下A549细胞上皮间质转化的影响并初步探讨其与脯氨酸4-羟化酶β亚基(P4HB)、波形蛋白(Vimentin)表达的相关性。方法 将A549细胞分为3组培养:常氧对照组(常规培养)，缺氧模型组(加入CoCl2培养模拟缺氧微环境诱导A549细胞上皮间质转化)，益气除痰方含药血清组(在CoCl2培养基础上加入益气除痰方含药血清培养)。相差显微镜下观察3组A549细胞形态的变化；RT-PCR检测3组549细胞P4HB、Vimentin mRNA的表达含量。结果 相差显微镜下常规培养的A549细胞呈不典型上皮细胞形态，经CoCl2刺激后，大部分细胞形态明显拉长，呈现明显的间质细胞形态。益气除痰方含药血清组与常氧对照组比较间质细胞形态的细胞增多，较缺氧模型组间质细胞形态的细胞明显减少。缺氧模型组A549细胞P4HB、Vimentin mRNA的表达量较常氧对照组明显上调，差异有统计学意义(P < 0.01)；益气除痰方含药血清组P4HB、Vimentin mRNA的表达量较缺氧模型组明显下调，差异有统计学意义(P < 0.01，P < 0.05)。结论 益气除痰方可抑制缺氧诱导的上皮间质转化，可能与其下调P4HB mRNA表达相关。

Objective To investigate the effects of Yiqi Huata Decoction(YHD)，a herbal recipe with the actions of invigorating qi and removing phlegm，on epithelial-mesenchymal transition(EMT) of A549 cells under the microenvironment of hypoxia，and to explore the correlation of EMT with prolyl 4-hydroxylase beta polypeptide(P4HB) expression. Methods Serum containing YHD was prepared. A549 cells were cultured under normal condition(normal group)，or co-cultured with CoCl2 to induce EMT(hypoxia model group)，or co-cultured with CoCl2 and serum containing YHD(medication group). Phase contrast microscopy was applied to observe the morphological change of A549 before and after medication. P4HB and Vimentin mRNA expression in A549 cells was detected by RT-PCR. Results Under phase contrast microscope，untreated A549 cells showed atypical features of epithelial cells，and after stimulation with CoCl2，most A549 cells became longer，showing obvious interstitial cell morphology. In medication group，cells with interstitial cell morphology were increased(P > 0.05 compared with the normal group)，while were decreased significantly in comparison of hypoxia model group(P < 0.05). The results of RT-PCR showed that P4HB and vimentin mRNA expression levels in hypoxia model group were 2.06±0.08 and 1.37±0.17，respectively，higher than 1.35±0.19 and 0.56±0.10 in normal group(P < 0.01). The expression level of P4HB was 0.87±0.10，and that of Vimentin mRNA was 0.96±0.17 in the medication group，lower than those in the hypoxia model group(P < 0.01，P < 0.05). Conclusion YHD shows inhibitory effect on hypoxia-induced EMT of A549 cells，which is probably related to the down-regulation of P4HB expression .

R285.5

国家自然科学基金资助项目( 81273963)；广东省自然科学基金重点项目(S2012020010886)；广东省自然科学基金(101510407010000021)。