摘要
目的:利用补中益气汤含药血清干预TGF-β介导的肺腺癌A549细胞,观察A549细胞中整合素β1的表达变化,探讨补中益气汤在肺癌侵袭和转移过程中的作用。方法:采用血清药理学方法制备含药血清,按照随机对照方法将40只SD大鼠随机分为4组,分别为补中益气汤高剂量含药血清组(11.34 g·kg-1)、补中益气汤中剂量含药血清组(5.67 g·kg-1)、补中益气汤低剂量含药血清组(2.84 g·kg-1)和空白血清组(10 m L·kg-1),每日给药一次,连续灌胃三天后提取大鼠血清,用于后续实验。将A549细胞分为空白血清组(10%大鼠空白血清)、TGF-β+空白血清组(10 ng·m L-1TGF-β+10%大鼠空白血清)、TGF-β+补中益气汤高剂量含药血清(10 ng·m L-1TGF-β+10%大鼠高剂量含药血清)、TGF-β+补中益气汤中剂量含药血清(10 ng·m L-1TGF-β+10%大鼠中剂量含药血清)、TGF-β+补中益气汤低剂量含药血清组(10 ng·m L-1TGF-β+10%大鼠低剂量含药血清)。MTT法检测补中益气汤含药血清对TGF-β介导的A549细胞的抑制率。细胞划痕实验检测补中益气汤含药血清对TGF-β介导的A549细胞迁移能力的影响。细胞免疫荧光技术、western blot方法检测整合素β1在蛋白水平上的表达。结果:与A549细胞空白血清组、TGF-β+空白血清组分别比较,补中益气汤高剂量、中剂量和低剂量含药血清可提高对TGF-β介导的A549细胞的抑制率(IR)(P<0.01),分别具有抑制TGF-β介导的A549细胞的迁移作用(P<0.01),且能减少整合素β1在蛋白水平上的表达(P<0.01)。结论:补中益气汤含药血清能抑制TGF-β介导的A549细胞的生长及迁移,以及能降低整合素β1在蛋白水平上的表达。
Objective ：To observe the Intergrin β1 of human lung adenocarcinoma cells A549 expression changes and to explore the effect of serum containing Buzhong Yiqi Decoction in invasion and metastasis of lung Adenocarcinoma cells A549 by using Buzhong Yiqi Decoction intervening the human lung adenocarcinoma cells A549 mediated by TGF - β. Methods ：The serum containing Buzhong Yiqi Decoction was prepared by adopting the method of serum pharmacology. 40 rats were divided into the four groups ： high （ 11.34 g · kg - 1 ）, middle （ 5.67 g · kg - 1 ）, low （ 2.84 g · kg -1 ） dose groups of Buzhong Yiqi Decoction and normal saline（ 10 mL · kg-1 ）. At the end of the 3thday, serum was obtained. A549 was divided into blank serum group（ 10% blank serum） ,TGF - β ＋ blank serum group（ 10 ng ·mL-1 TGF -β ＋ blank ser- um）, TGF - β ＋ high, middle and low dose groups of serum Buzhong Yiqi Decoction （ 10 ng· mL-1 TGF -β ＋ high, mid- dle,low of Buzhong Yiqi Decoction） Inhibition rate of Buzhong Yiqi Decoction on Integrinl31 of human lung adenocarci- noma cells A549 mediated by TGF - β was observed by MTT assay. Cellula immunofluorescence and Western blot were used to assay the protein expression of Integrin β1. Results： Compared with blank serum group, high,middle and low dose groups of serum Buzhong Yiqi Decoction can improve the inhibitiion rate（IR） of Buzhong Yiqi Decoction on Intergrin β1 of human tung adenocarcinoma cells A549 mediated by TGF - β significantly（ P 〈 0.01 ） and decrease the protein expression of Integrin β1 （P 〈 0. 01 ）. Conclusion：Buzhong Yiqi Decoction can improve the inhibition rate on Integrin β1 of human lung adenocarcinoma cells A549 mediated by TGF - β and reserve the invasion and metastasis of lung adenocarcinoma cells A549 mediated by TGF - β by decreasing the protein expression of Integrinβ1.
出处
《中华中医药学刊》
CAS
北大核心
2015年第10期2361-2364,I0005-I0007,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金项目(81473569)
关键词
补中益气汤
肺腺癌A549细胞
整合素Β1
侵袭
转移
Buzhong Yiqi Decoction
human lung adenocarcinoma cells A549
Integrin β1
invasion
metastasis
