基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响

刘珍财; 赵海霞; 马娜; 陈茜; 袁丁; 张长城

doi:10.13422/j.cnki.syfjx.20182330

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基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响

药理 | 更新时间：2024-09-23

- 基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响
- Effect of Wuzi Yanzong Fang on Proliferation of Spermatogonial Stem Cells in Aging Rats Through PI3K/Akt Signaling Pathway
- 中国实验方剂学杂志 2018年24卷第23期页码：119-125
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81573931，81503334，81774316，81373881）
- DOI：10.13422/j.cnki.syfjx.20182330
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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刘珍财, 赵海霞, 马娜, 等. 基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响[J]. 中国实验方剂学杂志, 2018,24(23):119-125.

LIU Zhen-cai, ZHAO Hai-xia, MA Na, et al. Effect of Wuzi Yanzong Fang on Proliferation of Spermatogonial Stem Cells in Aging Rats Through PI3K/Akt Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 119-125.
刘珍财, 赵海霞, 马娜, 等. 基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响[J]. 中国实验方剂学杂志, 2018,24(23):119-125. DOI： 10.13422/j.cnki.syfjx.20182330.

LIU Zhen-cai, ZHAO Hai-xia, MA Na, et al. Effect of Wuzi Yanzong Fang on Proliferation of Spermatogonial Stem Cells in Aging Rats Through PI3K/Akt Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 119-125. DOI： 10.13422/j.cnki.syfjx.20182330.

摘要

目的：探讨五子衍宗方（Wuzi Yanzong Fang，WYF）基于磷酯酰肌醇3-激酶（PI3K）/丝氨酸苏氨酸蛋白激酶（Akt）信号通路对老龄大鼠精原干细胞增殖的作用及其机制研究。方法：以自然衰老大鼠为研究对象，将40只18月龄SPF级SD雄性大鼠随机分为老龄模型组，WYF低、中、高剂量组，每组10只，另以10只2月龄大鼠作为青年组。WYF低、中、高剂量组分别灌胃给药0.4，0.8，1.6 g·kg-1，青年组、老龄模型组分别灌胃给予生理盐水，每周停药2 d，持续给药4个月。末次给药后麻醉处死大鼠，快速取出睾丸组织。免疫荧光法检测Sertoli细胞数量，实时荧光定量聚合酶链式反应（Real-time PCR）检测胶质细胞源性神经营养因子（GDNF），前髓细胞锌指蛋白（PLZF），干细胞因子（SCF）和骨形态蛋白4（BMP4）mRNA表达水平，免疫荧光法检测精原干细胞数量及增殖细胞核抗原（PCNA）的表达，蛋白免疫印迹法（Western blot）和免疫荧光法检测磷酯酰肌醇3-激酶（PI3K）和磷酸化丝氨酸苏氨酸蛋白激酶（p-Akt）的表达与定位。结果：与青年组比较，老龄模型组Sertoli细胞数量未见明显差异，而GDNF，PLZF，SCF和BMP4 mRNA表达水平显著降低，精原干细胞数量减少，PCNA表达下降，PI3K，p-Akt蛋白表达降低（P<0.01）；与老龄模型组比较，WYF各剂量组GDNF，PLZF，SCF和BMP4 mRNA表达水平显著升高，精原干细胞数量增多，PCNA表达上调，PI3K，p-Akt表达明显增多（P<0.05，P<0.01）。结论： WYF能够明显促进老龄大鼠精原干细胞的增殖，其作用机制可能与调节睾丸内PI3K/Akt信号通路有关。

Abstract

Objective:To investigate effect of Wuzi Yanzong Fang (WYF) on proliferation of spermatogonial stem cells in aging rats through phosphoinositol 3-kinase(PI3K)/protein kinase(Akt) signaling pathway. Method:The 18-month-old male SD rats were randomly divided into aging model group

and low

middle and high-dose WYF groups

with 10 rats in each group. Ten 2-month-old rats were used as young control group. low

middle and high-dose WYF groups were given WYF 0.4

0.8

1.6 g·kg-1 by gavage for 4 months

respectively. Young control group and aging model group were given normal saline

and the administration was suspended for 2 days a week. The administration lasted for 4 months. The rats were put to death after the final administration of WYF

and then the testes were quickly collected. The numbers of Sertoli cells were detected by immunofluorescence. The relative mRNA expression levels of glial cell line-derived neurotrophicfactor (GDNF)

promyelocyte Zinc finger protein (PLZF)

bone morphogenetic protein4 (BMP4) and stem cell factor (SCF) in testicular tissues were detected by Real-time quantitative PCR (Real-time PCR). The numbers of spermatogonial stem cells and the expression levels of PCNA were detected by immunofluorescence. The expressions and localizations of PI3K

Akt

p-Akt were detected by Western blot and immunofluorescence. Result:Compared with young control group

there was no significant difference in the numbers of Sertoli cells in each group

the mRNA expression levels of GDNF

PLZF

BMP4 and SCF in testis of aging model group were significantly decreased. The number of spermatogonial stem cells and levels of PI3K

p-Akt

PCNA expression decreased(P<0.01). After WYF treatment

the levels of GDNF

PLZF

BMP4 and SCF were significantly increased

the numbers of spermatogonial stem cells were increased

and PCNA

PI3K

p-Akt expressions were up-regulated(P<0.05

P<0.01). Conclusion:WYF can effectively promote the proliferation of spermatogonial stem cells in aging rats through PI3K/Akt signaling pathway.

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