Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele

LIANG Yichun; WANG Wen; LIANG Shulin; FU Ruijie; QIN Zhan; GENG Liguo; HE Chaoba; YUAN Shaoying

doi:10.3969/j.issn.1006-2157.2024.11.001

您当前的位置：

首页 >

文章列表页 >

Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele

Special Theme: TCM Prevention and Treatment of Male Diseases | 更新时间：2024-12-02

- Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele
- Journal of Beijing University of Traditional Chinese Medicine Vol. 47, Issue 11, Pages: 1481-1489(2024)
- 作者机构：
  
  1.广州中医药大学第二临床医学院　广州　510006
  2.广东省中医院珠海医院
- 作者简介：
  
  Prof.YUAN Shaoying, Master′s Degree, Chief Physician, Doctoral Supervisor. Zhuhai Hospital, Guangdong Provincial Hospital of Chinese Medicine, No.53, Jingle Road, Xiangzhou District, Zhuhai 510915.E-mail: ysy017@126.com
- 基金信息：
  
  Guangdong Provincial Administration of Traditional Chinese Medicine Traditional Chinese Medicine Research Project(202205111913416600)
- DOI：10.3969/j.issn.1006-2157.2024.11.001
  CLC： R285.5
- Received：23 May 2024，
  
  Published Online：10 October 2024，
  
  Published：30 November 2024
- 稿件说明：
移动端阅览
LIANG Yichun, WANG Wen, LIANG Shulin, et al. Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1481-1489.
DOI：

LIANG Yichun, WANG Wen, LIANG Shulin, et al. Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1481-1489. DOI： 10.3969/j.issn.1006-2157.2024.11.001.

摘要

目的

基于B淋巴细胞瘤－2(Bcl-2)/Bcl-2相关X蛋白(Bax)通路探讨活血养阴法改善精索静脉曲张大鼠生精功能的作用机制。

方法

选取8周龄SD雄性大鼠30只，按随机数字表法分为假手术组、模型组、左卡尼汀组(1.0 g/kg)、活血养阴法低剂量组(5.2 g/kg)、活血养阴法中剂量组(10.4 g/kg)及活血养阴法高剂量组(20.8 g/kg)，每组5只。除假手术组外，其余大鼠均通过经典的Turner法复制精索静脉曲张模型。术后4周，假手术组及模型组大鼠灌胃生理盐水，各给药组灌胃相应药物，每日1次，连续4周。灌胃结束后，使用计算机辅助精子分析仪检测精子质量，HE染色法观察睾丸组织病理学形态并通过Johnsen评分评价睾丸生精功能，TUNEL法检测睾丸细胞凋亡情况，实时荧光PCR法检测睾丸组织Bcl-2及Bax mRNA表达，免疫组织化学法检测细胞色素C蛋白表达。

结果

与假手术组比较，模型组大鼠的精子总数、精子活动率及(a＋b)级精子率均降低(

0.05)；睾丸组织中生精小管排列紊乱，有明显空泡样改变，Johnsen评分下降(

0.05)；睾丸组织细胞凋亡率增加(

0.05)；睾丸组织Bax mRNA表达上升(

0.05)；细胞色素C蛋白阳性表达增加(

0.05)。与模型组比较，活血养阴法中剂量组大鼠的精子总数，低、中剂量组精子活动率，以及低剂量组(a＋b)级精子率均上升(

0.05)；大鼠睾丸病理学结构有不同程度的改善，活血养阴法中剂量组Johnsen评分增加(

0.05)；左卡尼汀组与活血养阴法各剂量组睾丸细胞凋亡率均下降(

0.05)；活血养阴法低、高剂量组Bcl-2 mRNA表达上升(

0.05)，左卡尼汀组与活血养阴法中剂量组Bax mRNA表达下降(

0.05)；活血养阴法中剂量组细胞色素C蛋白阳性表达下降(

0.05)。

结论

活血养阴法可提高精索静脉曲张大鼠的精子质量，修复损伤的睾丸组织结构，改善生精功能，其作用机制可能与激活Bcl-2/Bax凋亡通路、抑制细胞凋亡有关。

Abstract

Objective

To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY) in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.

Methods

Thirty 8-week-old male SD rats were randomly divided into six groups (five rats per group): sham

model

levocarnitine (1.0 g/kg)

and PBCNY low-dose (5.2 g/kg)

PBCNY medium-dose (10.4 g/kg)

and PBCNY high-dose (20.8 g/kg) groups. Apart from the sham group

all the groups were built a varicocele model by the classical Turner method. After 4 weeks

the sham group and model group were gavaged with normal saline

and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks. At the end of gavage

the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer

and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score. TUNEL method was used to detect apoptosis

RT-qPCR was used to detect Bcl-2 and Bax mRNA expression

and immunohistochemical method was used to detect cytochrome C protein expression.

Results

Compared to the sham group

sperm total count

motility rate and percentage of sperm with grade (a+ b) motility were lower in the model group of rats (

0.05). HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score (

0.05). The result of TUNEL assay showed that the apoptosis rate was increased in the model group (

0.05). Bax mRNA expression was increased in testicular tissue (

0.05). The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group (

0.05). Compared with the model group

the rat sperm total count in the PBCNY medium-dose group

motility rate in the PBCNY low- and medium-dose groups

and percentage of sperm with grade (a+ b) motility in the PBCNY low-dose group increased(

0.05); the pathological structure of rat testis had different degrees of improvement

and Johnsen score increased in the PBCNY me

dium-dose group (

0.05); the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups (

0.05); the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups (

0.05)

and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group (

0.05); the positive expression of cytochrome C decreased in the PBCNY medium-dose group (

0.05).

Conclusion

The therapy of PBCNY can improve sperm quality

repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele

and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.

关键词

Keywords

references

RUSSO GI , SALEH R , FINOCCHI F , et al . Impact of varicocele on testicular oxidative stress and sperm parameters in experimental animals: a systematic review and meta-analysis [J ] . World J Mens Health , 2024 , 42 ( 3 ): 563 － 573 .

JENSEN CFS , STERGREN P , DUPREE JM , et al . Vari-cocele and male infertility [J ] . Nat Rev Urol , 2017 , 14 ( 9 ): 523 － 533 .

刘勋明，汪阳，袁少英 . 袁少英治疗精索静脉曲张合并不育症的经验 [J ] . 中医药导报， 2019 , 25 ( 19 ): 137 － 139 .

耿立果，袁少英，秦国政 . 丹红通精方对双侧精索静脉曲张术后精子质量的疗效观察 [J ] . 中华男科学杂志， 2021 , 27 ( 7 ): 635 － 640 .

游旭军，袁少英，何超拔，等 . 丹红通精方对精索静脉曲张不育症患者精液质量、缺氧诱导因子－1α的影响 [J ] . 中医药导报， 2017 , 23 ( 10 ): 72 － 74 .

TURNER TT . The study of varicocele through the use of animal models [J ] . Hum Reprod Update , 2001 , 7 ( 1 ): 78 － 84 .

李雷，严建燕，苏欣，等 . 大鼠睾丸组织固定法的优选研究 [J ] . 中国比较医学杂志， 2013 , 23 ( 6 ): 20 － 23 .

FU W , CUI J , TANG SS . The relationship of testicular stiffness with Johnsen score and sperm retrieval outcome in men with non-obstructive azoospermia [J ] . Quant Imaging Med Surg , 2024 , 14 ( 4 ): 3033 － 3043 .

GOLDBER E , SBERNA D , WHEAT TE , et al . Cytochrome c: immunofluorescent localization of the testis-specific form [J ] . Science , 1977 , 196 ( 4293 ): 1010 － 1012 .

FALCONE M , BOCU K , KESKIN H , et al . Anti-sperm antibody positivity in men with varicocele: a systematic review and meta-analysis [J ] . World J Mens Health , 2024 , 42 ： e43 .

TANAKA H , FUJISAWA M , TANAKA H , et al . Apoptosis-related proteins in the testes of infertile men with varicocele [J ] . BJU Int ， 2002 ， 89 ( 9 ): 905 － 909 .

WOOD GJA , CARDOSO JPG , PALUELLO DV , et al . Varicocele-associated infertility and the role of oxidative stress on sperm DNA fragmentation [J ] . Front Reprod Health , 2021 ， 3 : 695992 .

BUDZINSKA M , KAMIENICZNA M , WOJNAR L , et al . The role of the intrinsic pathway of apoptosis in human ejaculated sperm damage under a state of scrotal heat stress [J ] . J Assist Reprod Genet , 2024 , 41 ( 1 ): 99 － 108 .

ELAHI M , HOJATI V , HASHEMITABAR M , et al . Negative effect of varicocele on sperm mitochondrial dysfunction: a cross-sectional study [J ] . Int J Reprod Biomed , 2023 , 21 ( 4 ): 323 － 332 .

WANG HQ , ZHU BJ , JING T , et al . Lycopene inhibits apoptosis of mouse spermatocytes in varicocele via miR-23a/b-induced downregulation of PROK2 [J ] . Reprod Fertil Dev , 2024 , 36 : RD23136 .

NÖSSING C , RYAN KM . 50 years on and still very much alive: ′Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics′ [J ] . Br J Cancer , 2023 , 128 ( 3 ): 426 － 431 .

MOSTAFA T , RASHED L , NABIL N , et al . Seminal BAX and BCL2 gene and protein expressions in infertile men with varicocele [J ] . Urology , 2014 , 84 ( 3 ): 590 － 595 .

MOHAN S , ABDELWAHAB SI , KAMALIDEHGHAN B , et al . Involvement of NF-κB and Bcl2/Bax signaling pathways in the apoptosis of MCF7 cells induced by a xanthone compound Pyranocycloartobiloxanthone A [J ] . Phytomedicine , 2012 , 19 ( 11 ): 1007 － 1015 .

VUCICEVIC K , JAKOVLJEVIC V , COLOVIC N , et al . Association of Bax expression and Bcl2/Bax ratio with clinical and molecular prognostic markers in chronic lymphocytic leukemia [J ] . J Med Biochem , 2016 , 35 ( 2 ): 150 － 157 .

CAUWELIER C , RIDDER ID , BULTYNCK G . Recent advances in canonical versus non-canonical Ca 2＋－signaling-related anti-apoptotic Bcl-2 functions and prospects for cancer treatment [J ] . Biochim Biophys Acta Mol Cell Res , 2024 , 1871 ( 5 ): 119713 .

NGUYEN DH , SOYGUR B , PENG SP , et al . Apoptosis in the fetal testis eliminates developmentally defective germ cell clones [J ] . Nat Cell Biol , 2020 , 22 ( 12 ): 1423 － 1435 .

PESSOA J . Cytochrome c in cancer therapy and prognosis [J ] . Biosci Rep , 2022 , 42 ( 12 ): BSR20222171 .

LEWIS A , HAYASHI T , SU TP , et al . Bcl-2 family in inter-organelle modulation of calcium signaling; roles in bioenergetics and cell survival [J ] . J Bioenerg Biomembr , 2014 , 46 ( 1 ): 1 － 15 .

NEWTON K , STRASSER A , KAYAGAKI N , et al . Cell death [J ] . Cell , 2024 , 187 ( 2 ): 235 － 256 .

GARRIDO C , GALLUZZI L , BRUNET M , et al . Mechanisms of cytochrome C release from mitochondria [J ] . Cell Death Differ , 2006 , 13 ( 9 ): 1423 － 1433 .

WAGNER H , CHENG JW , KO EY . Role of reactive oxygen species in male infertility: an updated review of literature [J ] . Arab J Urol , 2018 , 16 ( 1 ): 35 － 43 .

刘海红，赵军，许瑾，等 . 对氧磷酶1基因rs662位点单核苷酸多态性与男性不育发病风险的相关性研究 [J ] . 中华男科学杂志， 2018 , 24 ( 8 ): 708 － 712 .

MOSTAFA T , RASHED L , NABIL N , et al . Seminal BAX and BCL2 gene and protein expressions in infertile men with varicocele [J ] . Urology , 2014 , 84 ( 3 ): 590 － 595 .

袁少英 . 精索静脉曲张合并男性不育的论治难点与对策 [J ] . 中国中西医结合杂志， 2013 , 33 ( 9 ): 1165 － 1167 .

耿立果，袁少英，覃湛，等 . 基于网络药理学及分子对接探讨丹红通精方治疗男性不育症的作用机制 [J ] . 中医学报， 2023 , 38 ( 5 ): 1061 － 1067 .

HAN XX , JIANG YP , LIU N , et al . Protective effects of Astragalin on spermatogenesis in streptozotocin-induced diabetes in male mice by improving antioxidant activity and inhibiting inflammation [J ] . Biomed Pharmacother , 2019 , 110 : 561 － 570 .

QIAN L . Modulation of cytokine level and sperm quality of mice by Lycium barbarum polysaccharides [J ] . Int J Biol Macromol , 2019 , 126 : 475 － 477 .

YU S , ZHAO Y , ZHANG FL , et al . Chestnut polysaccharides benefit spermatogenesis through improvement in the expression of important genes [J ] . Aging , 2020 , 12 ( 12 ): 11431 － 11445 .

ZHANG YJ , LIN F , YAN ZG , et al . Salidroside downregulates microRNA－133a and inhibits endothelial cell apoptosis induced by oxidized low-density lipoprotein [J ] . Int J Mol Med , 2020 , 46 ( 4 ): 1433 － 1442 .

吴泊，邵幼姿，薛莉丽，等．芽孢杆菌发酵炮制中药红花增强溶血栓药效研究 [J ] . 世界中医药， 2018 ， 13 ( 7 )： 1759 － 1762 .

YAO R , CAO Y , WANG CM , et al . Taohuajing reduces oxidative stress and inflammation in diabetic cardiomyopathy through the sirtuin 1/nucleotide-binding oligomerization domain-like receptor protein 3 pathway [J ] . BMC Complement Med Ther , 2021 , 21 ( 1 ): 78 .

Views

125

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency

Effect of therapeutic principle of "cultivating soil to restrict water" on ATP content and mitochondrial apoptosis pathway in rats with uremic cardiomyopathy

Protective effect of Honghua（Safflower,Flos Carthami） on acute liver injury induced by carbon tetrachloride in rats and mechanism study

Effects of Cigu Xiaozhi Wan on NF-κB mediated nonalcoholic fatty liver apoptosis

Effects of Yiqi Huoxue Fang on necrosis and apoptosis of myocardial cell in rats with myocardial infarction

Related Author

FAN Fangxin

ZHU Haoru

QIAN Meng

SHI Tuo

ZHU Sitai

XIE Ming

Minlin Zheng

Qin San

Related Institution

School of Traditional Chinese Medicine, Beijing University of Chinese Medicine

School of Integrative Medicine, Fujian University of Traditional Chinese Medicine

Research Institute of Integrative Medicine, Fujian University of Traditional Chinese Medicine

Inner Mongolia Medical University

Gansu University of Traditional Chinese Medicine

AI问答

Postal code：100029
Tel：(010)53911857 Email：jbutcm@bucm.edu.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰