Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice

Yinhu HUANG; Ziling WANG; Kunhang DU; Cheng WANG; Caihong HUANG; Ting YANG; Han WEI; Honghui JIANG; Lu WANG; Qinghua ZHANG; Yaping WANG

doi:10.16016/j.2097-0927.202401061

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Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice

Basic Medicine | 更新时间：2026-01-09

- Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice
- Journal of Army Medical University Vol. 46, Issue 10, Pages: 1123-1131(2024)
- 作者机构：
  
  1.400016 重庆，重庆医科大学：干细胞与组织工程研究室，组织学与胚胎学教研室
  2.400042 重庆，陆军特色医学中心妇产科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202401061
  CLC： R282.71;R339.21;R339.52
- Received：16 January 2024，
  
  Revised：2024-03-06，
  
  Published：30 May 2024
- 稿件说明：
移动端阅览
Yinhu HUANG, Ziling WANG, Kunhang DU, et al. Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice[J]. Journal of Army Medical University, 2024, 46(10): 1123-1131.
DOI：

Yinhu HUANG, Ziling WANG, Kunhang DU, et al. Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice[J]. Journal of Army Medical University, 2024, 46(10): 1123-1131. DOI： 10.16016/j.2097-0927.202401061.

摘要

目的

探讨人参皂苷Rg1减轻热应激致小鼠睾丸损伤的机制。

方法

20只6~8周龄雄性C57BL/6J小鼠按随机数字表法分为4组，每组5只。对照组(Control组)和热应激组(HS组)小鼠腹腔注射0.9%生理盐水[10 mL/(kg·d)×14 d]，热应激+人参皂苷Rg1组(HS+Rg1组)和人参皂苷Rg1组(Rg1组)小鼠腹腔注射人参皂苷Rg1[20 mg/(kg·d)×14 d]。HS组与HS+Rg1组小鼠在给药第7天时，将麻醉后小鼠下腹部置于43 ℃水浴中单次热应激30 min。小鼠精母细胞GC-2spd(ts)分为4组: 对照组(Control组)细胞常规培养48 h；热应激组(HS组)细胞常规培养36 h，置于43 ℃水浴中单次热应激30 min，继续培养12 h；热应激+Rg1组(HS+Rg1组)在细胞培养体系中加入Rg1(终浓度50 μmol/L)，热应激处理同HS组；Rg1组不给予热应激处理，其余同HS+Rg1组。建模完成后1 d采集眼球血，ELISA法检测血清睾酮水平；摘取睾丸观察形态并称量，计算睾丸指数，HE染色观察睾丸组织病理学形态；制备睾丸组织匀浆检测过氧化氢酶(CAT)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平；取附睾精子，计算机辅助精子分析(CASA)系统分析小鼠精子浓度和活力；TUNEL染色检测GC-2spd(ts)细胞凋亡情况；Western blot检测GC-2spd(ts)细胞Nrf2、Keap1、HO-1、Bax、Bcl-2和Caspase3蛋白表达量；RT-qPCR检测GC-2spd(ts)细胞Nrf2蛋白靶基因GCLC、GCLM和NQO1相对表达量。

结果

Rg1可阻止热应激所致睾丸质量和睾丸指数下降，减轻睾丸组织结构损伤，阻止精子浓度和活力下降，拮抗热应激所致睾丸间质细胞数量减少和血清睾酮水平下降，降低睾丸组织中MDA累积，提高抗氧化酶CAT和SOD的活性。Rg1还可减轻GC-2spd(ts)细胞凋亡，下调Bax、Caspase3和Keap1蛋白表达，增强Bcl-2、Nrf2和HO-1蛋白表达量，增强Nrf2蛋白靶基因GCLC、GCLM和NQO1相对表达量。

结论

Rg1对正常小鼠睾丸结构和功能影响不显著，但可有效提高小鼠睾丸抗热应激损伤能力，其机制可能与Rg1激活Nrf2信号通路，提高抗氧化酶活性，减少生精细胞凋亡有关。

Abstract

Objective

To explore the mechanism of ginsenoside Rg1 in alleviating heat stress-induced testicular injury in mice.

Methods

A total of 20 C57BL/6 male mice (6~8 weeks old) were randomly divided into 4 groups (

=5). The mice from the control group and heat stress (HS) group were intraperitoneally injected with 10 mL/(kg·d) 0.9 % normal saline for 14 d

while those in the HS+Rg1 group and the Rg1 group were given an intraperitoneal injection of 20 mg/(kg·d) for 14 d

and then on the 7th day after administration

the mice in the HS group and the HS+Rg1 group had the lower abdomen put into a 43 ℃ water bath for 30 min as a single heat stress after being anesthetized with 4% chloral hydrate. Mouse spermatocytes GC-2spd(ts) were divided into control group (routine culture for 48 h)

HS group (placed in a 43 ℃ water bath for 30 min after 36 h of conventional culture

and cultured till the end of 48 h)

HS+Rg1 group (50 μmol/L Rg1 treatment followed by heat stress injury)

and Rg1 group (no heat stress injury). In 1 d after modeling

the eyeball blood samples were collected to detect serum testosterone with ELISA

and the testicles were extracted to observe the morphology and weighed to calculate the testicular index

. HE staining was used to observe the histopathology of testis

and corresponding reagents and kits was employed to detect the content of malondialdehyde (MDA) and activities of catalase (CAT) and superoxide dismutase (SOD) in testis tissue. After the epididymal sperm were collected

the sperm concentration and motility were analyzed by computer-assisted sperm analysis (CASA) system. In

in vitro

experiments

cell apoptosis was detected with TUNEL staining

the protein levels of Nrf2

Keap1

HO-1

Bax

Bcl-2 and Caspase3 were detected with Western blotting

and the mRNA levels of GCLC

GCLM and NQO1 were detected by RT-qPCR.

Results

Rg1 prevented the decreases in testicular weight and testicular index caused by heat stress

reduced the damage of testicular tissue structure

prevented the decrease of sperm concentration and vitality

antagonized the decreasd number of Leydig cells and serum testosterone level

reduced the accumulation of MDA in testicular tissue

and enhanced the activities of CAT and SOD. Rg1 treatment alleviated the apoptosis of GC-2spd(ts) cells

down-regulated the expression of Bax

Caspase3 and Keap1 proteins

enhanced the expression of Bcl-2

Nrf2 and HO-1 proteins

and increased the transcriptional levels of Nrf2 target genes GCLC

GCLM and NQO1.

Conclusion

Rg1 has no significant effect on the structure and function of mouse testes

but it can effectively improve the ability of mouse testes to resist heat stress injury

which may be related to the activation of Nrf2 signaling pathway

the improvement of antioxidant enzyme activity

and the reduction of apoptosis of spermatogenic cells.

关键词

Keywords

references

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