主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (3): 200-207.doi: 10.3969/j.issn.1006-2157.2017.03.005

• 科技之窗 • 上一篇    下一篇

益气化痰散瘀方含药血清对低氧大鼠肺动脉平滑肌细胞凋亡的影响*

童祥丽1,朱洁2,杨程1,童佳兵1,李泽庚2#   

  1. 1 安徽中医药大学第一附属医院 安徽 230031;
    2 安徽中医药大学
  • 收稿日期:2016-01-03 出版日期:2017-03-30 发布日期:2017-03-30
  • 通讯作者: 李泽庚,男,教授,主任医师,博士生导师,研究方向:中医药防治呼吸系统疾病,E-mail:li6609@126.com
  • 作者简介:童祥丽,女,硕士,住院医师。
  • 基金资助:
    *国家自然科学基金项目(No.81373598,No.81403308)

Influence of medicated serum of Yiqi Huatan Sanyu Fang on apoptosis of pulmonary artery smooth muscle cells in hypoxia rats*

TONG Xiangli1, ZHU Jie2, YANG Cheng1, TONG Jiabing1, LI Zegeng2#
  

  1. 1 First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Anhui 230031;
    2 Anhui University of Traditional Chinese Medicine
  • Received:2016-01-03 Online:2017-03-30 Published:2017-03-30

摘要: 目的 观察益气化痰散瘀方含药血清对低氧大鼠肺动脉平滑肌细胞(PASMCs)凋亡的影响,并探讨其机制是否通过线粒体途径。方法 制备益气化痰散瘀方含药血清和空白血清。组织贴块法培养大鼠PASMCs并进行a-SMA 细胞免疫荧光鉴定。MTT法筛选最佳含药血清及最佳低氧时间:常氧对照组,低氧模型组,5%含药血清、10%含药血清、20%含药血清组(含药血清用空白血清稀释),分别作用12、24、48 h。正式实验根据MTT结果分为常氧对照组、低氧模型组、最佳含药血清组,含药血清作用最佳低氧时间后,TUNEL-FITC/DAPI法检测各组PASMCs凋亡情况,Western Blot法检测各组casepase-9、casepase-3、Bcl-2和Bax蛋白的表达。结果 MTT结果显示益气化痰散瘀方含药血清浓度为20%、低氧作用24 h时抑制作用最强,效果最佳。TUNEL-FITC/DAPI法显示与低氧模型组比较,益气化痰散瘀方含药血清可明显促进低氧条件下大鼠PASMCs的凋亡(P<0.01);Western Blot法显示益气化痰散瘀方含药血清可升高低氧大鼠PASMCs的casepase-9、casepase-3和 Bax蛋白表达(P<0.05,P<0.01),降低Bcl-2蛋白表达(P<0.01)。结论 益气化痰散瘀方含药血清可能通过线粒体途径诱导低氧大鼠PASMCs凋亡。

关键词: 益气化痰散瘀方, 含药血清, 肺动脉平滑肌细胞, 凋亡, 线粒体途径, 大鼠

Abstract: Objective To observe the influence of medicated serum of Yiqi Huatan Sanyu Fang on apoptosis of pulmonary artery smooth muscle cells (PASMCs) in hypoxia rats, and discuss the relationship between its mechanism and mitochondrial pathway. Methods The medicated serum of Yiqi Huatan Sanyu Fang and blank serum were prepared. PASMCs were cultured by using tissue explant method and given identification by using a-SMA immunofluorescence. After the best medicated serum and best hypoxia time were screened by using methyl thiazolyl tetrazolium test (MTT), PASMCs were divided into normoxia control group, hypoxia model group, 5% medicated serum group (low-dose group), 10% medicated serum group (mid-dose group) and 20% medicated serum group (high-dose group, medicated serum diluted by blank serum). The effective duration was, respectively, 12 h, 24 h and 48 h. PASMCs were divided, according to MTT results, into normoxia control group, hypoxia model group and best medicated serum group in formal test. The apoptosis of PASMCs was detected by using TUNEL-FITC/DAPI, and expressions of casepase-9, casepase-3, Bcl-2 and Bax protein were detected by using Western Blot in all groups after the effect on the best hypoxia time by medicated serum. Results The results of MTT showed that when the concentration of the medicated serum was 20% and hypoxia time was 24 h, the inhibitory effect was the highest with the best result. The results of TUNEL-FITC/DAPI showed that the medicated serum improved significantly the apoptosis of PASMCs compared with hypoxia model group (P<0.01) under hypoxia condition. The results of Western Blot showed that the medicated serum promoted the expressions of casepase-9, casepase-3 and Bax protein (P<0.05, P<0.01), and reduced expression of Bcl-2 protein (P<0.01). Conclusion The medicated serum of Yiqi Huatan Sanyu Fang can induce the apoptosis of PASMCs in hypoxia rats maybe through mitochondrial pathway.

Key words: Yiqi Huatan Sanyu Fang, medicated serum, pulmonary artery smooth muscle cells, apoptosis, mitochondrial pathway, rats

中图分类号: 

  • R285.5