补肾化瘀生新方改善缺血缺氧性微环境延缓骨髓间充质干细胞衰老的作用

张宝霞; 张金生; 惠小珊; 张丽娜

doi:10.13422/j.cnki.syfjx.20201604

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补肾化瘀生新方改善缺血缺氧性微环境延缓骨髓间充质干细胞衰老的作用

药理 | 更新时间：2023-07-12

- 补肾化瘀生新方改善缺血缺氧性微环境延缓骨髓间充质干细胞衰老的作用
- Mechanism of Bushen Huayu Shengxin Decoction in Delaying Senescence of Bone Marrow Mesenchymal Stem Cells by Improving Ischemic-hypoxic Microenvironment
- 中国实验方剂学杂志 2020年26卷第16期页码：87-92
- 作者机构：
  
  河南中医药大学第一附属医院，郑州 450000
- 作者简介：
  
  张宝霞，硕士，副主任医师，硕士生导师，从事针灸治疗心脑血管疾病基础与临床研究，E-mail：zhangbaoxiayy@126.com
  张金生，博士，主任医师，博士生导师，从事中医药防治心脑血管疾病基础与临床研究，E-mail：zjssir2004@sina.com.cn
- 基金信息：
  
  河南省科技攻关项目(192102310158);河南省中医药科学研究专项(2017ZY2065);国家自然科学基金面上项目(81373611)
- DOI：10.13422/j.cnki.syfjx.20201604
  中图分类号： R2-0;R22;R285.5
- 网络出版日期：2020-06-09，
  
  纸质出版日期：2020-08-20
- 稿件说明：
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张宝霞,张金生,惠小珊等.补肾化瘀生新方改善缺血缺氧性微环境延缓骨髓间充质干细胞衰老的作用[J].中国实验方剂学杂志,2020,26(16):87-92.

ZHANG Bao-xia,ZHANG Jin-sheng,HUI Xiao-shan,et al.Mechanism of Bushen Huayu Shengxin Decoction in Delaying Senescence of Bone Marrow Mesenchymal Stem Cells by Improving Ischemic-hypoxic Microenvironment[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(16):87-92.
张宝霞,张金生,惠小珊等.补肾化瘀生新方改善缺血缺氧性微环境延缓骨髓间充质干细胞衰老的作用[J].中国实验方剂学杂志,2020,26(16):87-92. DOI： 10.13422/j.cnki.syfjx.20201604.

ZHANG Bao-xia,ZHANG Jin-sheng,HUI Xiao-shan,et al.Mechanism of Bushen Huayu Shengxin Decoction in Delaying Senescence of Bone Marrow Mesenchymal Stem Cells by Improving Ischemic-hypoxic Microenvironment[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(16):87-92. DOI： 10.13422/j.cnki.syfjx.20201604.

摘要

目的

观察补肾化瘀生新方改善细胞生存的不同微环境对p16/pRb和p53/p21信号通路调控作用，探讨其对骨髓间充质干细胞（BMSCs)衰老的影响。

方法

采用无血清的1640培养基和低氧细胞工作站培养24 h建立缺血缺氧微环境细胞体外模型。随机分为空白组，模型组，补肾化瘀生新方组（24.58 g·kg

-1

），复合培养基组；复合培养基组加入复合培养基，模型组完全培养基培养；补肾化瘀生新方组加入含有补肾化瘀生新方的血清培养基，均在低氧细胞工作站培养24 h；空白组加入完全培养基正常培养；采用流式细胞仪检测BMSCs细胞周期、实时荧光定量聚合酶链式反应（Real-time PCR）检测p16

INK4a

，p53，p21 mRNA表达、及存活素（Survivn），半胱氨酸天冬氨酸蛋白酶-3（Caspase-3），聚腺苷二磷酸核糖聚合酶（PARP） mRNA表达；蛋白免疫印迹法（Western blot）检测

-catenin和糖原合成酶激酶-3

（GSK-3

）蛋白的表达。

结果

与空白组比较，模型组S期细胞数增多，G

期明显降低，p16

INK4a

，p53，p21，Survivn，Caspase-3，PARP mRNA表达明显升高，

-catenin和GSK-3

蛋白表达明显升高（

0.05）；与模型组比较，补肾化瘀生新方组S期细胞数减少，G

期比值明显升高（

0.05），p16

INK4a

，p53，p21，Survivn，Caspase-3，PARP mRNA表达明显降低，

-catenin和GSK-3

蛋白表达明显降低（

0.05）。

结论

补肾化瘀生新方通过改善缺血缺氧性微环境调控p16/pRb和p53/p21信号通路延缓BMSCs衰老。

Abstract

Objective

To explore the mechanism of Bushen Huayu Shengxin decoction in delaying senescence of bone mesenchymal stem cells(BMSCs) by improving cellular microenvironment and regulating p16/pRb and p53/p21 signaling pathways.

Method

The cells were cultured in serum-free 1640 medium and hypoxic cell workstation for 24 hours to establish the cell model of ischemic-hypoxic microenvironment in vitro

then randomized into control group (with complex medium)

model group (with complete medium)

and treatment group (with serum medium-containing Bushen Huayu Shengxin decoction)

and all were cultured in hypoxic cell workstation for 24 hours. The normal group was added with control culture for complete medium

The cell cycle of BMSCs was detected by flow cytometry

the expressions of p16

INK4a

p53

p21 and Survivn

cysteine aspartic acid protease-3 (Caspase-3)

polyadenosine diphosphate ribose polymerase (PARP) mRNA were analyzed by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR)

and the levels of

-catenin protein and glycogen synthase kinase-3

（GSK-3

） protein were detected by Western blot.

Result

Compared with the normal group

the proportion of S phase cells increased

while that at the G

phase decreased significantly in the model group (P

0.05). Compared with the model group

the proportion of S phase decreased

whereas that at the G

phase gradually increased in the treatment group (

0.05). Compared with the normal group

mRNA expressions of p16

INK4a

p53

p21 and Survivn

Caspase-3

PARP in the model group increased significantly (

0.05). Compared with the model group

mRNA expressions of p16

INK4a

p53

p21 and Survivn

Caspase-3

PARP in the treatment group decreased significantly (

0.05). Compared with the normal group

protein expressions of

-catenin and GSK-3

in the model group increased significantly (

0.05). Compared with the model group

protein expressions of

-catenin and GSK-3

in the treatment group decreased significantly (

0.05).

Conclusion

Bushen Huayu Shengxin decoction could delay the senescence of BMSCs by improving ischemic-hypoxic microenvironment and regulating p16/pRb and p53/p21 signaling pathways.

关键词

Keywords

references

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