化瘀祛痰方及其拆方对HepG2细胞胆固醇代谢的影响及机制研究

贾连群; 杨关林; 罗莹; 任路; 陈文娜; 冯峻屹; 崔勇

doi:10.13422/j.cnki.syfjx.2012.16.018

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化瘀祛痰方及其拆方对HepG2细胞胆固醇代谢的影响及机制研究

药理 | 更新时间：2024-09-23

- 化瘀祛痰方及其拆方对HepG2细胞胆固醇代谢的影响及机制研究
- Effect of Huayu Qutan Recipe and its Separated Recipes on Cholesterol Metabolism in HepG2 Cells
- 中国实验方剂学杂志 2012年18卷第16期页码：158-162
- 作者机构：
  
  1. 辽宁中医药大学
  2. 大连医科大学
- 作者简介：
- 基金信息：
  
  中国博士后科学基金特别项目（201104611）;中国博士后科学基金项目（20090451279）;辽宁省高等学校杰出青年学者成长计划项目（LJQ2011100）;辽宁省科技厅博士启动基金项目（20091052）
- DOI：10.13422/j.cnki.syfjx.2012.16.018
  中图分类号： R285.5
- 纸质出版日期：2012
- 稿件说明：
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[1]贾连群,杨关林,罗莹,任路,陈文娜,冯峻屹,崔勇.化瘀祛痰方及其拆方对HepG2细胞胆固醇代谢的影响及机制研究[J].中国实验方剂学杂志,2012,18(16):158-162.

JIA Lian-qun1, YANG Guan-lin1, LUO Ying2, et al. Effect of Huayu Qutan Recipe and its Separated Recipes on Cholesterol Metabolism in HepG2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2012, 18(16): 158-162.
[1]贾连群,杨关林,罗莹,任路,陈文娜,冯峻屹,崔勇.化瘀祛痰方及其拆方对HepG2细胞胆固醇代谢的影响及机制研究[J].中国实验方剂学杂志,2012,18(16):158-162. DOI： 10.13422/j.cnki.syfjx.2012.16.018.

JIA Lian-qun1, YANG Guan-lin1, LUO Ying2, et al. Effect of Huayu Qutan Recipe and its Separated Recipes on Cholesterol Metabolism in HepG2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2012, 18(16): 158-162. DOI： 10.13422/j.cnki.syfjx.2012.16.018.

摘要

目的:观察化瘀祛痰方药及其拆方对人肝癌细胞株HepG2胆固醇代谢的影响

探讨该方药的作用机制

并比较方药全方及各拆方的疗效及调控作用。方法:SD大鼠随机分为空白对照组、化瘀祛痰方全方组、补气组、化瘀组、祛痰组

生理盐水和相应中药予以ig

连续8 d。给药量为全方组17.9 g.kg-1

补气组7.5 g.kg-1

化瘀组5.4 g.kg-1

祛痰组4.2 g.kg-1

空白对照组ig等量生理盐水。第9天腹主动脉采血

分离血清。体外培养HepG2细胞

随机分为6组

即①空白对照组、②ox-LDL刺激组、③化瘀祛痰方全方组、④补气组、⑤化瘀组、⑥祛痰组。其中②组加入终质量浓度为50 mg.L-1ox-LDL

③～⑥组用相应含药血清（10%）预处理24 h后加入终质量浓度为50 mg.L-1ox-LDL

各组细胞培养24 h后进行各项指标测定。采用MTT法检测细胞活性;胆固醇氧化酶终点法检测HepG2细胞总胆固醇含量;反转录-聚合酶链反应（RT-PCR）定量分析3-羟基-3-甲基戊二酸单酰辅酶A还原酶（HMGCR）、B类Ⅰ型清道夫受体（SR-BⅠ）、胆固醇7α-羟化酶（CYP7A1）mRNA表达。结果:50 mg.L-1ox-LDL作用于HepG2细胞使其细胞活力显著降低至对照组的（59.37±18.21）%

细胞总胆固醇含量显著升高（幅度为71.52%）。而化瘀祛痰方全方、补气方、化瘀方、祛痰方显著升高细胞活力分别至（76.52±24.38）%

（68.13±20.35）%

（72.63±21.56）%

（73.92±23.47）%（P<0.05或P<0.01）

全方、化瘀方、祛痰方能显著降低HepG2细胞胆固醇含量

幅度分别为:28.77%

17.18%

25.09%。RT-PCR结果表明经ox-LDL诱导HepG2细胞HMGCR mRNA相对表达显著减少（P<0.01）

而SR-BⅠ和CYP7A1 mRNA相对表达显著增加（P<0.01

P<0.01）。化瘀祛痰方及祛痰方含药血清均能显著降低HepG2细胞HMGCR mRNA水平（P<0.01

P<0.05）

显著升高SR-BⅠ和CYP7A1mRNA水平（均P<0.01）。结论:化瘀祛痰方药全方及祛痰方可通过降低HMGCR水平和升高SR-BⅠ

CYP7A1水平从而影响胆固醇合成、摄取和转化

这可能是其调控脂质代谢发挥抗AS作用的机制之一。

Abstract

Objective:To observe the influence of Huayu Qutan recipe and its separated recipes on cholesterol metabolism-related genes in HepG2 cells.Method: SD rats were equally divided into five groups in random: the control

Huayu Qutan recipes

Buqi recipes

Huayu recipes and Qutan recipes groups.They were gastric perfused daily with normal saline and recipes for 8 d

respectively.Blood sample was collected from abdominal aorta at 2 h after ending administration on the 9th day

and the serum was separated for testing.The cultured HepG2 cells were divided into six groups randomly: the normal serum control

oxidized low-density lipoprotein（ox-LDL） stimulating

Huayu Qutan recipes

Buqi recipes

Huayu recipes and Qutan recipe groups.Then cell viability of HepG2 was detected by MTT method.The levels of total cholesterol in HepG2 cells were measured by a method of enzymatic reagents.RT-PCR was used to test the changes of expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase（HMGCR）

scavenger receptor class B type I（SR-BⅠ）and cholesterol 7α-hydroxylase（CYP7A1） genes.Result: After stimulating by 50 mg · L-1 ox-LDL

HMGCR mRNA level was obviously decreased while SR-BⅠand CYP7A1 mRNA levels were obviously increased in HepG2 cell（all P<0.01）.Compared with the ox-LDL stimulating group

the expression of HMGCR mRNA was significantly decreased while SR-BⅠand CYP7A1 mRNA significantly increased in the Huayu Qutan recipe group and Qutan recipe group.Conclusion: Huayu Qutan recipe and Qutan recipe can reduce the levels of total cholesterol in HepG2 cell.The change of cholesterol metabolism-related genes expression may be one of the mechanisms against AS.

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