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目的:探讨左/右归丸对小鼠精子功能的作用机制。方法:正常ICR雄性小鼠随机分为服用左/右归丸的实验组和服用生理盐水的对照组,实验组灌服含左/右归丸溶液,每次含左、右归丸各0.25 g,qd×14 d,每次1 ml,对照组以等量生理盐水代替。于末次灌胃后第7日与超促排卵的雌鼠合笼,检测到雌鼠阴栓后处死雌鼠,取1-细胞胚胎(受精卵),体外培养,观察卵裂率和囊胚形成率。另将上述确认让雌鼠受孕的雄鼠,于前次交配后3 d处死,取附睾精子,并行IVF,观察体外受精率和囊胚形成率,进一步观察与精子受精能力相关的完整顶体(acrosome-in-tact spermatozoa,AIS)率和顶体酶活性。结果:体内受精实验组的卵裂率(88.3±1.3%)高于对照组的(62.1±3.5%),差异有统计学意义(P=0.005)。实验组IVF受精率为82.4±2.9%,高于对照组的62.5±3.6%,差异有统计学意义(P=0.015)。不管体内受精还是体外受精,实验组与对照组的囊胚形成率均无统计学差异(P>0.05)。实验组顶体酶活性为44.29±5.49 IU,高于对照组的27.12±4.87 IU,差异有统计学意义(P=0.048),AIS率组间无统计学差异。结论:左/右归丸能提高小鼠精子功能,其作用机制有可能是通过调节顶体酶活性来影响小鼠精子功能。
Objective: To explore the mechanism of action of Zuogui Gui Wan on mouse sperm function. Methods: Normal ICR male mice were randomly divided into experimental group taking Zuogui Guiyuan pill and control group taking physiological saline. The experimental group was fed with Zuo / You Gui Wan solution containing 0.25 g , qd × 14d, each 1 ml, the control group with an equal volume of saline instead. On the 7th day after the last intragastric administration, the mice were caged with ovarian hyperstimulation and females were sacrificed after embolism of females were detected. The 1-cell embryos (fertilized eggs) were taken and cultured in vitro to observe the cleavage rate and blastocyst formation rate. In addition, the above-mentioned male mice which were confirmed to conceive female mice were sacrificed 3 days after the last mating, sperm of the epididymis was taken, IVF was performed, the rate of in vitro fertilization and the rate of blastocyst formation were observed, and the complete acrosome ( acrosome-in-tact spermatozoa, AIS) rate and acrosin activity. Results: The cleavage rate of the in vitro fertilization group (88.3 ± 1.3%) was significantly higher than that of the control group (62.1 ± 3.5%) (P = 0.005). IVF fertilization rate of experimental group was 82.4 ± 2.9%, which was higher than that of control group (62.5 ± 3.6%), the difference was statistically significant (P = 0.015). No difference was found in the blastocyst formation rate between the experimental group and the control group (P> 0.05), regardless of in vivo fertilization or in vitro fertilization. The acrosin activity in the experimental group was 44.29 ± 5.49 IU, which was higher than that in the control group (27.12 ± 4.87 IU) (P = 0.048). There was no significant difference between the two groups in the AIS rate. Conclusion: Zuo You You Pill can improve sperm function in mice, which may be through the regulation of acrosin activity to affect mouse sperm function.