金水六君煎及其拆方含药血清对人中性粒细胞弹性蛋白酶诱导A549细胞黏液高分泌的影响  

Effect of Jinshui Liujun Decoction and its Decomposed Prescription Serum on High Mucus Secretion In-duced by HNE in A549 Cells

作　　者：柏正平[1] 刘雨[1] 刘芳[2] 谭小宁[2] 谭电波[2] 邓秀娟[2] 李仲普[2] 刘俊[2] 侯公瑾 谭光波[2] Bai Zhengping;Liu Yu;Liu Fang(Hunan University of Chinese Medicine,Hunan,Changsha 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《中国中医急症》2019年第5期753-756,共4页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81473564);湖南省研究生科研创新项目(CX2017B423)

摘　　要：目的探讨金水六君煎及其拆方对A549细胞黏液高分泌模型黏蛋白MUC5AC、MUC5B表达的影响。方法制备金水六君煎及其拆方、生理盐水大鼠血清,将人肺腺癌细胞A549分为空白对照组、模型组、金水六君煎组、养阴组、化痰组。采用人中性粒细胞弹性蛋白酶(HNE)25 nmol/L诱导A549细胞黏液高分泌,20%空白大鼠血清及含药血清进行干预。CCK8法测定细胞活性,实时荧光定量PCR法(rt-PCR)检测细胞MUC5AC mRNA、MUC5B mRNA表达,Western blotting检测细胞MUC5AC mRNA、MUC5B mRNA表达。结果各组细胞活性无明显差异。与空白组比较,模型组细胞黏蛋白MUC5AC m RNA、MUC5B m RNA与蛋白表达明显增多(P <0.05或P <0.01)。与模型组比较,金水六君煎组及养阴组MUC5AC mRNA、MUC5B m RNA表达明显降低(P <0.05);金水六君煎组及化痰组MUC5AC m RNA、MUC5B m RNA表达明显降低(P <0.01)。结论人中性粒细胞弹性蛋白酶可诱导A549细胞粘蛋白高表达,成功建立黏液高分泌细胞模型。金水六君煎及其拆方含药血清可明显抑制黏蛋白MUC5AC mRNA、MUC5B mRNA与蛋白表达。Objective: To investigate the effects of Jinshui Liujun Decoction and its decomposition on the expression of mucin MUC5AC and MUC5B in A549 cell mucus hypersecretion model. Methods: Rat serum of Jinshui Liujun Decoction,its decomposed prescription and normal saline was prepared. Human lung adenocarcinoma A549 was divided into the blank control group,the model group,Jinshui Liujun group,Yangyin group and Huatan group. Human neutrophil elastase(HNE) 25 nmmol/L was used to induce high mucus secretion in A549 cells and 20% blank rat serum and drug-containing serum were used for intervention. The cell viability was determined by CCK8 method,the expression of MUC5AC mRNA and MUC5B mRNA was detected by real-time PCR(rt-PCR) and expression of MUC5AC mRNA and MUC5B mRNA protein was detected by Western blotting. Results: There was no significant difference in cell viability. Compared with the blank group,the expression of MUC5AC mRNA and MUC5B mRNA and proteins in the model group were significantly increased(P < 0.05 or P < 0.01). Compared with the model group,the gene expressions of MUC5AC mRNA and MUC5B mRNA in Jinshui Liujun group and Yangyin group were significantly decreased(P < 0.05);the expressions of MUC5AC mRNA and MUC5B mRNAproteins in Jinshui Liujun group and Huatan group were significantly decreased(P < 0.01). Conclusion: Human neutrophil elastase can induce high expression of mucin in A549 cells and successfully establish a mucus hypersecretory cell model. Jinshui Liujun Decoction and its decomposed prescription serum can significantly inhibit the expression of mucin MUC5AC mRNA and MUC5B mRNA both in genes and proteins.

关 键 词：金水六君煎 拆方 中性粒细胞弹性蛋白酶 含药血清 A549 黏液高分泌 

分 类 号：R285.5[医药卫生—中药学]