Transcriptomic Analysis of Wuzi Yanzongwan on Testicular Spermatogenic Function in Semi-castrated Male Mice
10.13422/j.cnki.syfjx.20240268
- VernacularTitle:经典名方五子衍宗丸干预半去势雄性小鼠生精功能的转录组学分析
- Author:
Dixin ZOU
1
;
Yueyang ZHANG
2
;
Xuedan MENG
3
;
Wei LU
4
;
Shuang LYU
5
;
Fanjun ZENG
6
;
Kun CHEN
7
;
Chang LIU
8
;
Zhongxiu ZHANG
1
;
Yu DUAN
1
;
Yihang DAI
3
;
Zhaoyi WANG
3
;
Zhimin WANG
1
;
Ruichao LIN
3
Author Information
1. Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences， Beijing 100700，China
2. Wangjing Hospital of China Academy of Chinese Medical Sciences， Beijing 100102，China
3. Chinese Medicine Quality Evaluation Center，Beijing Key Laboratory， School of Chinese Materia Medica，Beijing University of Chinese Medicine，Beijing 100102，China
4. The Affiliated Hospital of Inner Mongolia Medical University，Hohhot 010050，China
5. The First Hospital of Hebei Medical University，Shijiazhuang 050031，China
6. General Administration of Customs（Beijing） International Travel Health Care Center， Beijing 100013，China
7. Beijing Center for Vaccine Control，Beijing Institute for Drug Control， Beijing 102206，China
8. School of Sport Science，Beijing Sport University，Beijing 100084，China
- Publication Type:Journal Article
- Keywords:
famous classical formulas;
Wuzi Yanzongwan;
semi-castration;
spermatogenesis;
transcriptomics;
male infertility;
Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(1):61-69
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice， and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group， model group， testosterone propionate group（0.2 mg·kg-1·d-1， intramuscular injection） and Wuzi Yanzongwan group（1.56 g·kg-1·d-1， intragastric administration） according to body weight， with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function， while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention， hematoxylin-eosin（HE） staining was used to observe the histopathology of testis， enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of serum testosterone（T）， luteinizing hormone（LH） and follicle stimulating hormone（FSH）. The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group， the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened， and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to verify the transcriptome data. Through the annotation analysis of Gene Ontology（GO） and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes（KEGG）， the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group， the testicular tissue of mice in the model group showed spermatogenic injury， contraction and vacuolization of the seminiferous tubules， reduction of spermatogenic cells at all levels， widening of the interstitial space， obstruction of spermatogonial cell development and other morphological abnormalities， and serum T significantly decreased， LH significantly increased（P<0.01）， and FSH elevated but no statistically significant difference， the count and vitality of epididymal sperm significantly decreased（P<0.01）. There were 882 differentially expressed mRNAs in the testicular tissues， of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group， the damage to testicular tissue in the Wuzi Yanzongwan group was reduced， the structure of the seminiferous tubules was intact， vacuolization was reduced， and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased， LH significantly decreased（P<0.01）， and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased（P<0.01）. Moreover， Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice， of which 32 were up-regulated and 127 were down-regulated， and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis， renewal， differentiation， metabolism， apoptosis and signal transduction of spermatogenic cells， and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function， endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice， and its mechanism may be related to the regulation of testicular transcriptional regulatory network， the synthesis of sex hormones in testicular interstitial cells， the function of spermatogenic stem cells， the whole cell cycle process of spermatogenesis， as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.
