Effect of Bushen Huayu Shengxin Formula Combined with Intravenous Injection of Bone Marrow Mesenchymal Stem Cells in Delaying Senescence of Rat

ZHANG Bao-xia; ZHANG Jin-sheng; XU Er-ping; HUI Xiao-shan; ZHANG Li-na

doi:10.13422/j.cnki.syfjx.20182482

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Effect of Bushen Huayu Shengxin Formula Combined with Intravenous Injection of Bone Marrow Mesenchymal Stem Cells in Delaying Senescence of Rat

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- Effect of Bushen Huayu Shengxin Formula Combined with Intravenous Injection of Bone Marrow Mesenchymal Stem Cells in Delaying Senescence of Rat
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 24, Pages: 116-121(2018)
- 作者机构：
  
  1. 河南中医药大学第一附属医院
  2. 河南中医药大学第三附属医院
  3. 河南中医药大学
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20182482
  CLC： R285.5
- Published：2018
- 稿件说明：
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[1]张宝霞,张金生,许二平,惠小珊,张丽娜.补肾化瘀生新法协同静脉注射骨髓间充质干细胞对延缓大鼠衰老作用[J].中国实验方剂学杂志,2018,24(24):116-121.

ZHANG Bao-xia, ZHANG Jin-sheng, XU Er-ping, et al. Effect of Bushen Huayu Shengxin Formula Combined with Intravenous Injection of Bone Marrow Mesenchymal Stem Cells in Delaying Senescence of Rat[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 116-121.
[1]张宝霞,张金生,许二平,惠小珊,张丽娜.补肾化瘀生新法协同静脉注射骨髓间充质干细胞对延缓大鼠衰老作用[J].中国实验方剂学杂志,2018,24(24):116-121. DOI： 10.13422/j.cnki.syfjx.20182482.

ZHANG Bao-xia, ZHANG Jin-sheng, XU Er-ping, et al. Effect of Bushen Huayu Shengxin Formula Combined with Intravenous Injection of Bone Marrow Mesenchymal Stem Cells in Delaying Senescence of Rat[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 116-121. DOI： 10.13422/j.cnki.syfjx.20182482.

摘要

目的:观察补肾化瘀生新法协同静脉注射骨髓间充质干细胞（bone mesenchymal stem cells

BMSCs）延缓大鼠衰老的有效性

明确中医药协同静脉注射BMSCs治疗延缓机体衰老的可行性。方法:75只SD大鼠随机分为补肾化瘀生新方高、中、低剂量组、模型组和正常组。每组12只

按照125 mg·kg

-1

·d

-1

的剂量颈背部皮下每日1次注射D-半乳糖溶液

连续45 d

建立大鼠衰老模型。补肾化瘀生新方高、中、低剂量组(27.28

13.64

6.82 g·kg

-1

)灌胃给药

模型组生理盐水灌胃

正常组不给药

早晚各1次

连续45 d。同时尾静脉注射BMSCs注射3.0×10

个/m L的BMSCs

每周1次

连续4次。免疫荧光法测定脑、肝、肺、心中CD105

p53

p21

p16

INK4a

阳性细胞数

实时荧光定量聚合酶链式反应（Real-time PCR）检测大鼠的脑、肝、肺、心中p16

INK4a

p53

p21 mRNA表达。结果:与正常组比较

模型组大鼠脑、心脏、肺脏、肝脏中CD105阳性细胞数明显降低

p16

INK4a

p21

p53阳性细胞数明显升高（P

0.05）

大鼠的脑、心脏、肺脏、肝脏中p16

INK4a

p53

p21 mRNA表达明显上调（P

0.05）;与模型组比较

补肾化瘀生新各剂量组大鼠脑、心脏、肺脏、肝脏中CD105阳性细胞数明显升高

p16

INK4a

p53

p21阳性细胞数明显降低（P

0.05）

补肾化瘀生新各剂量组均能明显降低脑、肝、肺、心组织中p16

INK4a

p53

p21阳性细胞数mRNA表达明显下降（P

0.05）。结论:补肾化瘀生新法协同静脉注射BMSCs可能影响p16

INK4a

/Rb和p53/p21途径上的关键基因p16

INK4a

p53

p21的表达而延缓BMSCs衰老

显示了中医药协同静脉注射BMSCs抗衰防衰的可行性。

Abstract

Objective:To observe the effect of Bushen Huayu Shengxin formula combined with intravenous injection of bone marrow mesenchymal stem cells（BMSCs）in delaying senescence of rat

so as to define the feasibility of traditional Chinese medicine combined with intravenous BMSCs in delaying aging of human body.Method:Totally 75 SD rats were randomly divided into high

medium and low-dose drug groups

the control group

the model group

with 12 rats in each group.The rats received a subcutaneous injection of D-galactose solution with the dose of 125 mg·kg

-1

·d

-1

once a day

for 45 d to establish rat aging models.The drug groups were administered with the Bushen Huayu Shengxin formula in 27.28

13.64

6.82 g·kg

-1

respectively in the morning and evening for 45 d（high-dose group

mid-dose group and low-dose group）.The model group was administered with normal saline

and the control group was not given any drug.At the same time

every rat was injected with 3.0×10

/m L BMSCs through tail vein once a week for four weeks.Immumofluorescence method was used to detect the number of CD105

P53

P21

p16

INK4a

-positive cells in brain

liver

lung

spleen

kidney

heart.Real-time fluorescent PCR（Real-time PCR）was used to detect the expressions of CD105

P53

P21

p16

INK4a

.Result:Compared with the normal group

the expression of CD105 in brain

liver

lung

spleen

kidney

heart was significantly lower

while P53

P21

p16

INK4a

were significantly higher in the model group（P

0.05）

mRNA expressions of P53

P21

p16

INK4a

in brain

liver

lung

spleen

kidney

heart were significantly up-regulated（P

0.05）.Compared with the model group

the number of CD105 in brain

liver

lung

spleen

kidney

heart was significantly higher

while P53

P21

p16

INK4a

-positive cells were significantly lower in high

medium and low-dose drug groups（P

0.05）

drug groups significantly decreased mRNA expressions of P53

P21

p16

INK4a

-positive cells in brain

liver

lung

spleen

kidney

heart（P

0.05）.Conclusion:Bushen Huayu Shengxin formula combined with intravenous injection of BMSCs may delay the aging of BMSCs by effecting the expressions of key genes p16

INK4a

p53

p21 on the pathways of p16

INK4a

/Rb and p53/p21.It shows that traditional Chinese medicine combined with intravenous BMSCs on delaying aging of human body is feasible.

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Related Author

ZHANG Jin-sheng

ZHANG Bao-xia

XU Er-ping

HUI Xiao-shan

ZHANG Li-na

ZHAO Hongli

ZHAO Xiaoxuan

MA Jing

Related Institution

Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University

The Third Clinical School of Zhejiang Chinese Medical University

Hebei University of Chinese Medicine

Hebei Key Laboratory of Integrated Traditional Chinese and Western Medicine for Lung Disease Research

Tianjin University of Traditional Chinese Medicine

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