Effect of Wuzi Yanzong Fang in Promoting Spermatogonial Stem Cells Proliferation in Aging Rats by Regulating miR-let-7-Imp Axis
10.13422/j.cnki.syfjx.20190333
- VernacularTitle: 五子衍宗方通过调节miR-let-7-Imp轴对老龄大鼠精原干细胞增殖的影响
- Author:
Qian CHEN
1
;
Hai-xia ZHAO
1
;
Na MA
1
;
Zhen-cai LIU
1
;
Ding YUAN
2
;
Chang-cheng ZHANG
1
Author Information
1. Medical College of China Three Gorges University, Yichang 443002, China
2. Renhe Hospital of China Three Gorges University, Yichang 443001, China
- Publication Type:Research Article
- Keywords:
Wuzi Yanzong Fang;
spermatogonial stem cells;
aging;
miR-let-7-imp
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(3):45-52
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the effect and mechanism of Wuzi Yanzong Fang (WZ) on proliferation of spermatogonial stem cells in aging rats by regulating miR-let-7-Imp axis.Method: A total of 40 18-month-old male SD rats were randomly divided into aging model group, and low, middle and high-dose WZ groups, with 10 rats in each group. Ten 2-month old rats were used as young control group. Low, middle and high-dose WZ groups were given by gastric WZ 0.4, 0.8, 1.6 g·kg-1 respectively. Young control group and aging model group were given normal saline for 4 months, which was suspended for 2 days every week. Rats were put to death after the final treatment with WZ, and then the testes were quickly removed from rats. The relative mRNA expression levels of miR-let-7 and Imp were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The expressions and localization of p-JAK2/JAK2, phosphate(p)-signal transduction and transcriptional activators3 (STAT3)/STAT3 signaling pathway proteins were detected by Western blot and immunofluorescence. The numbers of spermatogonial stem cells and the expression levels of proliferating cell nuclear antigen (PCNA) were detected by immunofluorescence. The testicular tissue morphology was observed using htoxylin eosin (HE) staining.Result: Compared with young control group, the expression levels of miR-let-7 mRNA were significantly increased, while the expression levels of Imp mRNA were decreased in the aging model group(P<0.01). Conversely, the expression levels of miR-let-7 mRNA were significantly decreased (P<0.05, P<0.01), while the mRNA expression levels of Imp mRNA were significantly increased after WZ treatment(P<0.05). The results of Western blotting and immunofluorescence showed that the expression levels of p-JAK2, p-STAT3 in aging model group were decreased significantly(P<0.01), while WZ significantly increased the expression levels of p-JAK2, p-STAT3(P<0.05, P<0.01). Immunofluorescence results showed that compared with young control group, the number of spermatogonial stem cells and PCNA expression decreased in aging model group. After WZ treatment, the number of spermatogonial stem cells was increased, whereas PCNA expression was up-regulated. HE showed that WZ significantly improved the testicular tissue structure of aging rats.Conclusion: WZ effectively promote the proliferation of spermatogonial stem cells by regulating miR-let-7-imp axis in testis.
