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1.贵州医科大学 基础医学院,贵阳 550025;
2.广州中医药大学 继续教育学院,广州 510405
秦臻,博士,副教授,从事中医药防治心血管疾病的研究,Tel:0851-88174021,E-mail:qinzhen@gmc.edu.cn
黄水清,博士,教授,博士生导师,从事中医药防治心脑血管疾病的研究,Tel:021-36585505,E-mail:hsq@gzucm.edu.cn
收稿日期:2018-12-28,
网络出版日期:2019-05-05,
纸质出版日期:2019-08-20
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秦臻, 韦正新, 黄水清. 当归补血汤对低流体剪切应力作用下内皮细胞功能损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(16):12-16.
Zhen QIN, Zheng-xin WEI, Shui-qing HUANG. Protective Effect of Danggui Buxue Tang on Impaired Functional Activity of Endothelial Cells Exposed to Low Flow Shear Stress[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(16): 12-16.
秦臻, 韦正新, 黄水清. 当归补血汤对低流体剪切应力作用下内皮细胞功能损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(16):12-16. DOI: 10.13422/j.cnki.syfjx.20191602.
Zhen QIN, Zheng-xin WEI, Shui-qing HUANG. Protective Effect of Danggui Buxue Tang on Impaired Functional Activity of Endothelial Cells Exposed to Low Flow Shear Stress[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(16): 12-16. DOI: 10.13422/j.cnki.syfjx.20191602.
目的:
2
观察不同当归、黄芪质量比(归芪比)的当归补血汤对低流体剪切应力(FSS)损伤内皮细胞功能的保护作用。
方法:
2
采用平行平板流动小室加载低FSS进行造模,实验将内皮细胞分为空白组和模型组(分别予以M199培养液培养2 h),辛伐他汀组(予以0.1 μmol·L
-1
的辛伐他汀培养2 h),当归补血汤组(予以3 g·L
-1
归芪比1∶1,1∶3,1∶5的当归补血汤培养2 h),培养结束后弃上清,空白组加载正常FSS,余各组加载低FSS,在30,60,360 min时分别收集每组细胞及灌流液,噻唑蓝(MTT)比色法检测细胞增殖力,硝酸酶还原法检测灌流液中一氧化氮(NO)含量,实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测细胞一氧化氮合酶(eNOS) mRNA及蛋白的表达。
结果:
2
与空白组比较,模型组细胞分泌NO及表达eNOS在60 min时显著升高(
P
<
0.01),随后在360 min降低;与模型组比较,当归补血汤对细胞增殖功能无明显影响,但可明显促进第360 min时细胞的NO分泌及eNOS表达,以归芪比1∶3,1∶5的效果更为显著。
结论:
2
当归补血汤对低FSS作用下的内皮细胞功能损伤具有一定的保护作用。
Objective:
2
To investigate the protective effect of Danggui Buxue Tang(DGBX)with Angelicae Sinensis Radix(AS) and Astragali Radix(AR)at different radios on impaired functional activity of endothelial cells(ECs)exposed to low-fluid shear stress (FSS).
Method:
2
Low FSS was loaded by a parallel plate flow chamber
and ECs were divided into normal FSS group
low FSS group(each preincubated with M199 medium for 2 h)
simvastatin group(preincubated with 0.1 μmol·L
-1
simvastatin for 2 h)
and 3 DGBX groups(preincubated with 3 g·L
-1
AS and AR at 1∶1
1∶3
1∶5 for 2 h
respectively). Then
the normal group was exposed to 1.2 Pa FSS
while the rest groups were all exposed to low FSS. At time points of 30
60
360 min
the proliferation was detected by methyl thiazoly tetrazolium(MTT)
the secretion of nitric oxide(NO) was detected by nitrase reduction test
and the mRNA and protein expressions of endothelial nitric oxide synthase(eNOS) were detected by Real-time fluorescence guantitative polymerase chain reaction(Real-time PCR) and Western blot
respectively.
Result:
2
Compared with the normal group
the secretion of NO and the expression of eNOS in ECs were both increased significantly at 60 min (
P
<
0.01)
then decreased at 360 min. Compared with the model group
there was no significant change in proliferation in DGBX groups
but DGBX could promote the NO secretion and the expression of eNOS in ECs exposed to low FSS at 360 min respectively
whereas the function of DGBX(AS and AR at 1∶3
1∶5)was obviously observed.
Conclusion:
2
DGBX could protect the functional activity of ECs exposed to low FSS.
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