摘要
目的:探究锁阳对少、弱精子症大鼠睾丸重量、血清睾酮浓度、以及精子数量、活动率等精子参数的影响,及促进未分化精原细胞增殖的机制,为开发新的治疗男性少、弱精子症的中药提供实验和理论依据。方法:选取8周龄,体重约(220±10)g的SD大鼠30只,随机分为5组(n=6):空白对照组、模型对照组、3组实验组(低、中、高锁阳浓度剂量);将模型对照组与锁阳实验组予以环磷酰胺[30 mg/(kg·d)]腹腔注射,连续5 d,构建大鼠少、弱精子症模型;将3组实验组分别用低浓度[0.5 g/(kg·d)]、中浓度[1 g/(kg·d)]及高浓度[2 g/(kg·d)]锁阳水煎液灌胃,每天1次,连续4周后观察各组精子数量、活动率,测量各组大鼠血清睾酮浓度,并取各组大鼠睾丸称重;采用Real-time PCR方法检测低浓度组睾丸组织中精原干细胞标志物(Oct4、Thy1、PLZF、C-kit)表达情况,采用β-actin作为内参;采用Real-time PCR方法检测低浓度组睾丸组织中胶质细胞源性神经营养因子(GDNF)表达情况。结果:空白对照组、模型对照组及低、中、高浓度锁阳实验组的大鼠睾丸重量分别为(1.52±0.06)g、(1.55±0.06)g、(1.34±0.04)g、(1.35±0.40)g、(1.43±0.30)g,不同浓度锁阳实验组大鼠睾丸重量与空白对照组、模型对照组比较未见明显差异(P>0.05)。空白对照组、模型对照组及低、中、高浓度实验组在每10个高倍镜视野精子数(精子数/10HP)分别为200±15、134±30、216±25、196±5、202±20个,锁阳实验组与模型对照组比较可显著提高大鼠的精子数量(P<0.05);低浓度锁阳水提物组睾丸组织中Oct4、Thy1、PLZF、GDNF基因mRNA表达水平较模型对照组增加,差异有统计学意义(P<0.05),C-kit基因mRNA表达水平未见明显差异(P<0.05);空白对照组、模型对照组及低、中、高浓度锁阳实验组的大鼠每10个高倍镜视野精子活动率(精子活动率/10HP)分别为(52.1±5.5)%、(38.1±2.5)%、(49.6±1.0)%、(58.7±9.5)%、(59.1±9.5)%;大鼠血清睾酮浓度分别为(190.0±87.5)、(82.5±25.8)、(185.0±82.4)、(331.0±86.7)、(229.0±75.6)mmol/L,锁阳实验组与模型对照组比较可显著提高大鼠的精子活动率及血清睾酮浓度(P均<0.05),但与空白对照组比较无明显差异(P>0.05)。结论:锁阳能显著提高少、弱精子症大鼠的精子数量、精子活动能力、血清睾酮水平,其改善机制可能是:1通过诱导睾丸Sertoli细胞中GDNF表达,促进未分化精原细胞增殖,从而促进精子发生过程,增加附睾尾部精子数目;2通过促进睾酮分泌,提高血清睾酮水平,进而改善精子活动率。
Objective： To investigate the effects of cynomorium songaricum （CS） decoction on the testis weight, serum testos- terone level, and sperm parameters of rats with oligoasthenospermia （ OAS）, explore its action mechanism of improving the proliferation of undifferentiated spermatogonial cells, and provide some experimental and theoretical evidence for the development of new Chinese drugs for OAS. Methods： Thirty 8-week-old male SD rats were randomly divided into five groups of equal number： blank control, model control, high-dose CS, medium-dose CS, and low-dose CS. OAS models were established by intraperitoneal injection of cyclo- phosphamide and, a month later, treated intragastrically with normal saline or CS at 2, 1, and 0.5 g per kg of the body weight per day, all for 4 weeks. Then, the testes of the animals were harvested to obtain the testicular weight, sperm concentration and motility, and the level of serum testosterone （T）, detect the expressions of the transcription factor I （Oct4）, Thy-1 cell surface antigen （Thyl）, promyelocytic leukemia zinc finger （PLZF）, KIT proto-oncogene receptor tyrosine kinase （C-kit） and glial cell-derived neu- rotrophic factor （GDNF） in the testis tissue of the rats in the low-dose CS group by real-time PCR. Results ： The testis weights in the blank control, model control, high-dose CS, medium-dose CS, and low-dose CS groups were （ 1.52 ±0.06）, （ 1.55 ±0.06）, （ 1.43 ± 0.30）, （ 1.35 ± 0.40） and （ 1.34 ± 0.04） g, respectively, not significantly different in the blank and model controls from those in the CS groups （P 〉 0.05）. The visual field sperm count per 10 HP was significantly increased in the high-, medium-, and low-dose CS groups （202 ± 20, 196 ± 5 and 216 ± 25） as compared with the blank and model controls （200 ± 15 and 134 ± 30） （ P 〈 0.05 ）. The mRNA expressions of the Oct4, Thyl, PLZF and GDNF genes were remarkably higher in the low-dose CS group than in the controls （ P 〈 0.05 ）, but that of the C-kit gene showed no significant difference from the latter （ P 〉 0.05 ）. The visual field sperm motility per 10 HP was markedly increased in the blank control （ [ 52. 1 ± 5.5 ] % ）, model control （ [ 38. 1 ± 2.5 ] % ）, high-dose CS （ [ 59.1 ± 9.5 ] % ）, medium-dose CS （ [ 58.7 ± 9.51% ）, and low-dose CS （ [ 49.6 ± 1.0 ] % ） groups, and so was the level of ser- um testosterone （ [ 190 ± 87.5], [ 82.5 ± 25.8 ], [ 229 ± 75.6 ], [ 331 ± 86.7 ] and [ 185 ± 82.4 ] retool/L）, both remarkably higher in the CS groups than in the model controls （ P 〈 0.05 ） but with no statistically significant difference between the CS groups and the blank controls （ P 〉 0.05 ）. Conclusion ： CS can significantly improve sperm concentration, sperm motility and serum T level in OAS rats, probably by inducing the expression of GDNF in the rat Sertoli cells, promoting the proliferation of undifferentiated spermatogonial cells, and enhancing spermatogenesis.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2016年第12期1116-1121,共6页
National Journal of Andrology
基金
徐州市卫生创新(科技攻关)团队(XWCX201603)
江苏省重点研发项目(BE2016641)~~
关键词
锁阳水煎液
少、弱精子症
精子活动率
精子数量
血清睾酮
未分化精原细胞
胶质细胞源性神经营养因子
cynomorium songaricum
oligoasthenospermia
sperm motility
sperm count
serum testosterone
undifferentiated spermatogonial cell
glial cell-derived neurotrophic factor
