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锁阳改善少弱精子症大鼠的精子数量、活力及血清睾酮水平并促进未分化精原细胞增殖

[Cynomorium songaricum improves sperm count and motility and serum testosterone level and promotes proliferation of undifferentiated spermatogonia in oligoasthenospermia rats].

作者信息

Cao Yi-Juan, Li Zhen-Bei, Qi Yu-Juan, Liu Ying, Gu Juan, Hu Fang-Fang, Zhang Wen-da, Hao Lin, Hou Jian-Quan, Han Cong-Hui

机构信息

Department of Urology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, China.

Center of Reproductive Medicine, Xuzhou Central Hospital, Xuzhou, Jiangsu 221009, China.

出版信息

Zhonghua Nan Ke Xue. 2016 Dec;22(12):1116-1121.

Abstract

OBJECTIVE

To investigate the effects of cynomorium songaricum (CS) decoction on the testis weight, serum testosterone 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 cyclophosphamide 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 1 (Oct4), Thy-1 cell surface antigen (Thy1), promyelocytic leukemia zinc finger (PLZF), KIT proto-oncogene receptor tyrosine kinase (C-kit) and glial cell-derived neurotrophic 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, Thy1, 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±9.5]%), and low-dose CS ([49.6±1.0]%) groups, and so was the level of serum testosterone ([190±87.5], [82.5±25.8], [229±75.6], [331±86.7] and [185±82.4] mmol/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).

CONCLUSIONS

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.

摘要

目的

探讨锁阳水煎剂对少弱精子症(OAS)大鼠睾丸重量、血清睾酮水平及精子参数的影响,探究其改善未分化精原细胞增殖的作用机制,为OAS新药研发提供实验及理论依据。

方法

将30只8周龄雄性SD大鼠随机分为5组,每组数量相等:空白对照组、模型对照组、高剂量锁阳组、中剂量锁阳组和低剂量锁阳组。通过腹腔注射环磷酰胺建立OAS模型,1个月后,分别用生理盐水或每天按2、1和0.5 g/kg体重的剂量灌胃给予锁阳水煎剂,共给药4周。然后,摘取动物睾丸以获取睾丸重量、精子浓度和活力以及血清睾酮(T)水平,采用实时荧光定量PCR检测低剂量锁阳组大鼠睾丸组织中转录因子1(Oct4)、Thy-1细胞表面抗原(Thy1)、早幼粒细胞白血病锌指蛋白(PLZF)、KIT原癌基因受体酪氨酸激酶(C-kit)和胶质细胞源性神经营养因子(GDNF)的表达。

结果

空白对照组、模型对照组、高剂量锁阳组、中剂量锁阳组和低剂量锁阳组的睾丸重量分别为(1.52±0.06)、(1.55±0.06)、(1.43±0.30)、(1.35±0.40)和(1.34±0.04)g,空白对照组和模型对照组与锁阳组相比差异无统计学意义(P>0.05)。高、中、低剂量锁阳组每10个高倍视野的视野精子计数显著高于空白对照组和模型对照组(分别为202±20、196±5和216±25)(200±15和134±30)(P<0.05)。低剂量锁阳组Oct4、Thy1、PLZF和GDNF基因的mRNA表达显著高于对照组(P<0.05),但C-kit基因的表达与对照组相比差异无统计学意义(P>0.05)。空白对照组([52.1±5.5]%)、模型对照组([38.1±2.5]%)、高剂量锁阳组([59.1±9.5]%)、中剂量锁阳组([58.7±9.5]%)和低剂量锁阳组([49.6±1.0]%)每10个高倍视野的视野精子活力显著升高,血清睾酮水平([190±87.5]、[82.5±25.8]、[229±75.6]、[331±86.7]和[185±82.4]mmol/L)也显著升高,锁阳组均显著高于模型对照组(P<0.05),但与空白对照组相比差异无统计学意义(P>0.05)。

结论

锁阳可显著提高OAS大鼠的精子浓度、精子活力和血清T水平,可能是通过诱导大鼠支持细胞中GDNF的表达,促进未分化精原细胞的增殖,增强精子发生。

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