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(PPPPPPPConclusion: WZ effectively promote the proliferation of spermatogonial stem cells by regulating miR-let-7-imp axis in testis.
ABSTRACT
Objective To study protective effects of Wuzi Yanzong Fang on DNA damage induced by cyclophosphamide (CTX) in mice, and explore its mechanism. Methods BalB/c mice were randomly divided into normal group, model group, Wuzi Yanzong Fang low dose group and Wuzi Yanzong Fang high dose group. Mice in Wuzi Yanzong Fang groups were pretreated with Wuzi Yanzong Fang for 7 days, then the mice in Wuzi Yanzong Fang groups and model group were intraperitoneally injected with CTX (100 mg/kg) every other day for three times, and mice in Wuzi Yanzong Fang groups were continued administered with Wuzi Yanzong Fang. Animals were sacrificed in twelve hours after the final treatment of CTX. ELISA was used to detect 8-OHdG content in serum, and single cell gel electrophoresis to detect DNA damage in bone marrow cells. Results Wuzi Yanzong Fang low dose group and high dose group reduced the level of 8-OHdG in serum. Wuzi Yanzong Fang significantly decreased Olive tail moment, tail moment, tail length and tail DNA%in mouse bone marrow cells. Conclusion Wuzi Yanzong Fang has good protective effects on DNA damage caused by CTX.