Lai Yongwei, Xi Yanli, Shao Minghai, Cui Xiaoli, Wei Xuemiao, Li Lele, Wang Yanchun, Fan Hongyan
Jilin Medical University, Jilin 132013, China.
Wei Sheng Yan Jiu. 2020 Sep;49(5):790-794. doi: 10.19813/j.cnki.weishengyanjiu.2020.05.017.
To explore the effect of myrica flavone on male reproductive toxicity induced by cyclophosphamide and its mechanism.
Thirty 6-week-old male ICR mice were randomly divided into 5 groups: blank control group, cyclophosphamide reproductive injury model group, myricetin low-medium high-dose intervention group. Except the blank control group, the other groups were intraperitoneally injected with cyclophosphamide 50 mg/kg daily for 7 consecutive days. The myricetin group received intragastric administration of 100, 200, and 400 mg/kg myricetin daily for 30 consecutive days since the second day of modeling. The blank control group and the model control group were given an equal volume of a 0. 25% sodium carboxymethyl cellulose solution. The body weight was measured every 3 days, and the day after the last administration, the mice were sacrificed by cervical dislocation, and the epididymis and testes were quickly taken. Testicular weighing, testicular index calculation, epididymis to obtain sperm, sperm analyzer to analyze sperm density and vitality. The expression of Bax and Bcl-2 in testicular tissues was detected by immunoblotting, and the mitochondrial membrane potential of sperm was detected by flow cytometry.
After 9 days of modeling, the weight of mice in the model group was lower than that of the blank control group, which was statistically different(P<0. 05). There was no difference between the myricetin treatment group and the model group. The testis index of the model group was(3. 93±0. 91)mg/g, which was significantly lower than that of the blank control group(6. 93±0. 98)mg/g, and the difference was statistically significant(P<0. 05). After treatment with bayberry flavonoids, the testis index increased, in the 100 and 200 groups and 400 mg/kg testis index were(3. 94±1. 21) mg/g, (4. 33±0. 88) mg/g, and(4. 80±0. 43) mg/g, respectively. Compared with model control group, The difference was statistically significant(P<0. 05 and P<0. 01). Compared with the control group, the sperm density, sperm rate of forward movement, sperm rate of non-forward movement, and decreased sperm rate of non-moving sperm increased in the model group. After treatment with bayberry flavonoids, compared with the model group, the sperm density, sperm rate of forward motion, and sperm rate of non-forward motion increased, and the immobility sperm rate decreased. The 200 and 400 mg/kg groups had statistical significance(P<0. 05 or P<0. 01); the normal rate of sperm mitochondrial membrane potential in the model group was(54. 70±5. 45)%, and the normal mitochondrial membrane potential rate after treatment with myricetin of 100, 200 and 400 mg/kg(59. 10±9. 97)%, (62. 10±6. 07)% and(77. 10±8. 87)%, of which the 400 mg/kg group was statistically significant(P<0. 05); the ratio of Bax/Bcl-2 in the model group was 5. 92±1. 45, and the ratio of Bax/Bcl-2 decreased after treatment with myricetin of 100, 200 and 400 mg/kg, which were 2. 52±0. 51, 1. 71±0. 52 and 1. 07±0. 29. There were statistical differences(P<0. 05 or P<0. 01).
Myrica flavone can protect sperm mitochondrial membrane potential, inhibit testicular cell apoptosis, and protect the male mice from reproductive toxicity induced by cyclophosphamide.
探讨杨梅黄酮对环磷酰胺所致雄性生殖毒性的影响及其作用机制。
将30只6周龄雄性ICR小鼠随机分为5组：空白对照组、环磷酰胺生殖损伤模型组、杨梅素低中高剂量干预组。除空白对照组外，其余各组连续7天每日腹腔注射环磷酰胺50mg/kg。自造模第2天起，杨梅素组连续30天每日灌胃给予100、200和400mg/kg杨梅素。空白对照组和模型对照组给予等体积的0.25%羧甲基纤维素钠溶液。每3天测量一次体重，末次给药后次日，颈椎脱臼处死小鼠，迅速取出附睾和睾丸。进行睾丸称重、计算睾丸指数，取附睾获取精子，用精子分析仪分析精子密度和活力。采用免疫印迹法检测睾丸组织中Bax和Bcl-2的表达，采用流式细胞术检测精子的线粒体膜电位。
造模9天后，模型组小鼠体重低于空白对照组，差异有统计学意义（P<0.05）。杨梅素治疗组与模型组之间无差异。模型组睾丸指数为（3.93±0.91）mg/g，显著低于空白对照组（6.93±0.98）mg/g，差异有统计学意义（P<0.05）。经杨梅黄酮治疗后，睾丸指数升高，100、200mg/kg组和400mg/kg组睾丸指数分别为（3.94±1.21）mg/g、（4.33±0.88）mg/g和（4.80±0.43）mg/g。与模型对照组相比，差异有统计学意义（P<0.05和P<0.01）。与对照组相比，模型组精子密度、前向运动精子率、非前向运动精子率及不动精子率均降低。经杨梅黄酮治疗后，与模型组相比，精子密度、前向运动精子率及非前向运动精子率升高，不动精子率降低。200和400mg/kg组有统计学意义（P<0.05或P<0.01）；模型组精子线粒体膜电位正常率为（54.70±5.45）%，100、200和400mg/kg杨梅素治疗后精子线粒体膜电位正常率分别为（59.10±9.97）%、（62.10±6.07）%和（77.10±8.87）%，其中400mg/kg组有统计学意义（P<0.05）；模型组Bax/Bcl-2比值为5.92±1.45，100、200和400mg/kg杨梅素治疗后Bax/Bcl-2比值降低，分别为2.52±0.51、1.71±0.52和1.07±0.29。差异有统计学意义（P<0.05或P<0.01）。
杨梅黄酮可保护精子线粒体膜电位，抑制睾丸细胞凋亡，对环磷酰胺所致雄性小鼠生殖毒性具有保护作用。
