Effect of Cooling Blood Method in Inhibiting Macrophage  Apoptosis Against Atherosclerosis

Xue-qian LIU; Jing WANG; Shou-pei CAO; Yao-hong SONG

doi:10.13422/j.cnki.syfjx.20190336

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Effect of Cooling Blood Method in Inhibiting Macrophage Apoptosis Against Atherosclerosis

Pharmacology | 更新时间：2021-02-09

- Effect of Cooling Blood Method in Inhibiting Macrophage Apoptosis Against Atherosclerosis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 3, Pages: 59-65(2019)
- 作者机构：
  
  1.南京中医药大学，南京　 210023
  2.南京中医药大学　附属南京中医院，南京　 210001
  3.南京市红十字医院，南京　 210001
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190336
  CLC： R2-0;R22;R285.5
- Received：09 July 2018，
  
  Published Online：16 November 2018，
  
  Published：05 February 2019
- 稿件说明：
移动端阅览
Xue-qian LIU, Jing WANG, Shou-pei CAO, et al. Effect of Cooling Blood Method in Inhibiting Macrophage Apoptosis Against Atherosclerosis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 59-65.
DOI：

Xue-qian LIU, Jing WANG, Shou-pei CAO, et al. Effect of Cooling Blood Method in Inhibiting Macrophage Apoptosis Against Atherosclerosis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 59-65. DOI： 10.13422/j.cnki.syfjx.20190336.

摘要

目的:

观察凉血散瘀法清心通脉饮含药血清对乙酰化低密度脂蛋白(acetylated low density lipoprotein

ac-LDL)诱导的小鼠单核巨噬细胞系RAW264.7凋亡率及A型清道夫受体(type A scavenger receptor

SR-A)，B细胞淋巴瘤-2(B-cell lymphoma-2，Bcl-2)，Bcl-2相关X蛋白(Bcl-2-associated X protein

Bax)，肌醇需要酶1

(inositol-requiring enzyme l

，IRE1

)表达的影响，探讨清心通脉饮治疗动脉粥样硬化可能的作用机制。

方法:

8只新西兰兔采用随机数字法分为阿托伐他汀组(2.6 g·kg

-1

)，清心通脉饮低、中、高剂量组(3.33，6.66，13.32 mg·kg

-1

)，灌胃7 d后颈动脉采血收集含药血清。各组用2.5，5，10，20%体积分数的含药血清培养液分别刺激RAW264.7细胞系6，12，24 h，采用细胞增殖与活性检测-8(CCK-8)法观察细胞增殖率。体外培养RAW264.7细胞系，分为空白组、模型组、阿托伐他汀组、清心通脉饮低、中、高剂量组。空白组用牛血清白蛋白(BSA)培养细胞，模型组用BSA+ 50 mg·L

-1

ac-LDL刺激细胞24 h，其他组用BSA+ 50 mg·L

-1

ac-LDL+ 10%含药血清刺激细胞24 h。流式细胞技术检测每组RAW264.7细胞系凋亡率和SR-A的表达，蛋白免疫印迹法(Western blot)检测Bcl-2，Bax

IRE1

蛋白的表达。

结果:

与空白组比较，模型组可显著增加RAW264.7细胞凋亡率(

0.01)，并增加Bax

SR-A蛋白表达(

0.01)，减少Bcl-2蛋白表达(

0.05)。与模型组比较，清心通脉饮低、中、高剂量组均可降低RAW264.7细胞系凋亡率(

0.05)，减少SR-A和IRE1

表达(

0.05，

0.01)。清心通脉饮低、高剂量组可降低Bax表达(

0.05，

0.01)；清心通脉饮中、高剂量组可降低Bcl-2表达(

0.05)。

结论:

清心通脉饮可降低巨噬细胞凋亡率，其治疗动脉粥样硬化机制可能与调控相关凋促亡蛋白Bax

IRE

，SR-A和抗凋亡蛋白Bcl-2的表达有关。

Abstract

Objective:

To observe the effect of serum-containing Qingxin Tongmai decoction(QXTMD) on the apoptosis rate of mouse mononuclear macrophage cell line RAW264.7 induced by Acetylated low density lipoprotein (ac-LDL) and the expressions of type A scavenger receptor(SR-A)

B-cell lymphoma-2(Bcl-2)

Bcl-2-associated X protein(Bax)

inositol-requiring enzyme 1

(IRE1

)

exploring the possible mechanism of QXTMD in the treatment of atherosclerosis.

Method:

Eight New Zealand rabbits were randomly divided into the atorvastatin group (2.6 g·kg

-1

) low

medium and high-dose QXTMD groups (3.33

6.66

13.32 mg·kg

-1

). After 7 days of gavage

the carotid blood was collected to prepare drug-containing serum. The RAW264.7 cell line was stimulated with 2.5%

10%

and 20%drug-containing serum culture for 6

and 24 h

respectively. The cell proliferation rate was observed by cell counting kit-8 (CCK-8) method. The RAW264.7 cell line was cultured

in vitro

and divided into blank group

model group

atorvastatin group

and low

medium and high-dose QXTMY groups. The cells in blank group were cultured with bovine serum albumin(BSA). The model group was stimulated with BSA+ 50 mg·L

-1

ac-LDL for 24 h. The other groups were stimulated with BSA+ 50 mg·L

-1

ac-LDL+ 10%drug-containing serum for 24 h. The apoptosis rate and SR-A expression of RAW264.7 cells were detected by flow cytometry. The expressions of Bcl-2

Bax and IRE1

protein were detected by Western blot.

Result:

Compared with the blank group

the model group could increase the apoptosis rate of RAW264.7 cells (

0.01) and the expressions of Bax and SR-A protein (

0.01)

but decrease the expression of Bcl-2 protein (

0.05). Compared with the model group

low

medium and high-dose QXTMD groups could decrease the apoptosis rate of RAW264.7 cell line (

0.05) and the expressions of SR-A and IRE1

(

0.05

0.01). The low-dose QXTMD group and the high-dose QXTMD group could decrease the expression of Bax(

0.05

0.01). The middle-dose group and the high-dose group could decrease the expression of Bcl-2(

0.01).

Conclusion:

QXTMD can reduce the apoptosis rate of macrophages. The mechanism of atherosclerosis may be related to the expressions of Bax

IRE

SR-A and anti-apoptotic protein Bcl-2.

关键词

Keywords

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