目的 探討五子衍宗丸對少弱精子癥小鼠生精功能、六磷酸肌醇激酶1（IP6K1）表達(dá)及線粒體DNA（mtDNA）甲基化的影響。方法 將雄性BALB/c小鼠隨機(jī)分為對照組、模型組及五子衍宗丸（1、2、4 g/kg）組，每組10只。除對照組外，其他各組單次ip 40 mg/kg白消安。從造模第2天開始，各組按劑量ig給藥，給藥4周后計算睪丸和附睪臟器系數(shù)，檢測精子數(shù)量和活力，蘇木精–伊紅（HE）染色觀察小鼠睪丸病理組織學(xué)改變，TUNEL染色觀察睪丸細(xì)胞凋亡情況，RT-qPCR、Western blotting和免疫組化（IHC）染色檢測IP6K1表達(dá)，Western blotting檢測DNA甲基轉(zhuǎn)移酶1（DNMT1）表達(dá)，焦磷酸測序檢測mtDNA D-loop甲基化，RT-qPCR檢測線粒體基因表達(dá)。結(jié)果 五子衍宗丸可恢復(fù)少弱精子癥小鼠的睪丸和附睪系數(shù)，顯著提高精子數(shù)量和活力，明顯改善睪丸組織病理學(xué)損傷和生精細(xì)胞的凋亡，顯著促進(jìn)IP6K1的表達(dá)，顯著抑制DNMT1表達(dá)和線粒體D-loop甲基化水平，顯著上調(diào)線粒體編碼基因的表達(dá)（P＜0.5、0.1、0.001）。結(jié)論 五子衍宗丸可改善少弱精子癥小鼠的生精功能，其機(jī)制可能與促進(jìn)IP6K1表達(dá)，抑制DNMT1/mtDNA D-loop甲基化，進(jìn)而促進(jìn)線粒體編碼基因的表達(dá)有關(guān)。

Objective To investigate the effects of Wuzi Yanzong Pills on spermatogenesis, expression of inositol hexaphosphate kinase 1 (IP6K1) and methylation of mitochondrial DNA (mtDNA) in oligoasthenospermia mice. Methods Male BALB/c mice were randomly divided into control group, model group, and Wuzi Yanzong Pills (1, 2, and 4 g/kg) group, with 10 mice in each group. Except the control group, the other groups were given 40 mg/kg intraperitoneal injection. Starting from the second day of model making, each group was given the drug by intragathy according to dose. After 4 weeks of administration, the coefficients of testis and epididymis organs were calculated, the sperm number and vitality were detected, the histopathological changes of testis were observed by HE staining, and the apoptosis of testicle cells was observed by TUNEL staining. The expression of IP6K1 was detected by RT-qPCR, Western blotting and immunohistochemistry (IHC). The expression of DNA methyltransferase 1 (DNMT1) was detected by Western blot, the D-loop methylation of mtDNA was detected by pyrosequencing, and the mitochondrial gene expression was detected by RT-qPCR. Results Wuzi Yanzong Pills could partially restore the testicular and epididymal coefficients of oligoasthenospermia mice, significantly increase sperm count and motility, significantly improve testicular histopathological injury and spermatogenic cell apoptosis, and significantly promote the expression of IP6K1. DNMT1 expression and mitochondrial D-loop methylation levels were significantly inhibited, and the expression of mitochondrial coding genes was significantly up-regulated (P < 0.5, 0.1, 0.001). Conclusion Wuzi Yanzong Pills can improve the spermatogenic function of oligoasthenospermia mice, and its mechanism may be related to promoting the expression of IP6K1, inhibiting DNMT1/mtDNA D-loop methylation, and promoting the expression of mitochondrial coding genes.

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天津市衛(wèi)健委、天津市中醫(yī)藥管理局中醫(yī)中西醫(yī)結(jié)合科研項目（2021071，2021072）