摘要
目的探讨化瘀祛痰方药及其拆方含药血清对脂多糖(LPS)诱导人脐静脉内皮细胞EA.hy926 TLR4/NF-κB信号通路活化的干预作用。方法 40只SD大鼠随机分为5组,即空白对照组、全方组、补气组、化瘀组、祛痰组,各组大鼠分别以生理盐水和相应中药煎剂连续灌胃9 d,末次灌胃给药2 h后,腹主动脉采血,离心后分离血清。体外培养EA.hy926细胞,随机分为7组,即①正常对照组、②LPS刺激组、③全方组、④补气组、⑤化瘀组、⑥祛痰组、⑦空白血清对照组。其中②组加入终浓度为10μg/ml的LPS,③～⑦组用各组含药血清(浓度为10%)预处理24 h后加入终浓度为10μg/ml的LPS,各组细胞培养24 h后进行各项指标测定。采用实时定量反转录-聚合酶链反应(Real-time PCR)定量分析TLR4、NF-κB和TNF-αmRNA表达;ELISA法检测TNF-α含量;Western印迹检测TLR4、NF-κB蛋白表达。结果与正常对照组相比,LPS刺激后TLR4、NF-κB和TNF-α表达显著增加(P<0.01),全方组TLR4、NF-κB和TNF-α的表达与刺激组相比显著减少(P<0.01);拆方各组中,化瘀组TLR4、NF-κB和TNF-α水平均显著减少(P<0.01),而补气组和祛痰组TLR4、NF-κB和TNF-α水平降低不明显。结论化瘀祛痰方药及化瘀拆方可显著抑制内皮细胞TLR4/NF-κB炎症信号通路的活化,这可能是其抗AS的作用机制之一。
Objective To observe the influence of Huayuqutan recipe and its separated recipes on TLR4/NF-κB signal pathway in cultured human umbilical veins endothelial cells EA. hy926 induced by LPS. Methods 55 SD rats were equally divided into five groups in random： the control, Huayuqutan, Buqi, Huayu, Qutan recipes groups. They were gastric perfused daily with normal saline and decoction respectively. Blood drawn from abdominal aorta of rats 2 h after ending perfusion on the 9th day, and the serum separated （drug-serum） was taken for testing. EA. hy926 cells were cultured equally divided into seven groups in random： the normal control, LPS stimulation, Huayuqutan, Buqi, Huayu, Qutan recipes and normal serum control groups. Then the mRNA expressions of TLR4, NF-κB and TNF-α were tested by real-time PCR. Content of TNF-α was detected by ELISA, while protein expressions of TLR4 and NF-κB were tested by Western-blot method. Results The expressions of TLR4, NF-κB and TNF—α were increased compared to normal group after stimulation of LPS （P 〈 0.01 ）. The expressions of TLR4, NF-κB and TNF—α in Huayuqutan recipe group and Huayu recipe group were obviously inhibited compared to LPS stimulation group （P 〈 0. 01 ）. Conclusions The molecular mechanism of Huayulutan recipe and Huayu recipe against AS is probably to inhibit the activation of TLR4/NF-κB signal pathway.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2012年第23期5159-5161,共3页
Chinese Journal of Gerontology
基金
国家自然科学基金青年基金项目(81202834)
中国博士后科学基金项目(No.201104611)
辽宁省高等学校杰出青年学者成长计划资助项目(LJQ20111100)
沈阳市科技局计划项目(F12-277-1-49)
关键词
化瘀祛痰
动脉粥样硬化
炎症信号通路
Huayuqutan recipe
Atherosclerosis
Inflammation signal pathway
