KLHL15基因对雄性小鼠睾丸和肝脏的影响

doi:10.6040/j.issn.1671-7554.0.2022.1324

摘要/Abstract

摘要： 目的探讨KLHL15基因缺失对雄性小鼠睾丸、肝脏组织结构和功能的影响。方法比较野生型(WT)和KLHL15敲除型(KLHL15^-/^-)雄性小鼠的睾丸、肝脏组织在结构和功能上的差异,测定指标包括:(1)生育能力:子代生育数、精子活力及精子质量;(2)解剖学:比较生长情况相近的野生型、KLHL15^-/^-雄性小鼠睾丸、肝脏质量及脏器系数;(3)病理组织学:通过HE观察 KLHL15基因缺失对小鼠肝脏、睾丸结构的影响;(4) 肝功能:原位末端标记法(Tunel)检测肝脏组织细胞凋亡情况;基于串联质量标签(TMT)标记定量蛋白组学分析方法对肝脏差异蛋白进行富集分析。结果 (1)生育能力:与WT雄性小鼠比较,KLHL15^-/^-雄性小鼠精子总数减少,不运动精子(IM)数量增加,精子测定参数平均曲线运动速度(VCL)、平均路径速度VAP降低(P=0.010, P=0.020);(2)解剖学:与WT小鼠比较,KLHL15^-/^-小鼠肝脏、睾丸的质量降低(P<0.001, P=0.026),脏器系数降低(P<0.001);(3)病理组织学:KLHL15^-/^-小鼠睾丸曲细精管管腔变大,生精细胞损失较多,上皮结构明显变薄,精子数目减少;肝脏细胞排列紊乱,肝血窦扩张;(4)肝功能:Tunel结果显示,KLHL15^-/^-小鼠肝脏出现更多凋亡细胞;富集分析结果显示, KLHL15基因缺失影响小鼠肝脏中线粒体和各种细胞器膜的组成,影响过氧化物酶体参与的反应以及多种氨基酸代谢。结论 KLHL15基因的缺失可破坏睾丸结构,并能降低雄性小鼠的精子活力、精子质量而导致其生育力减弱;可导致小鼠肝脏结构损伤,影响其代谢功能。

关键词: KLHL15基因, 敲除小鼠, 精子质量, 肝脏, 睾丸

Abstract: Objective To explore the effects of KLHL15 gene deletion on the structure and function of testis and liver in male mice. Methods The structure and function of testis and liver tissues of wild type(WT)and KLHL15 knockout type(KLHL15^-/^-)male mice were compared. The measurement indexes included: (1) Fertility: number of offspring, sperm vitality and sperm quality; (2) Anatomy: the mass and organ index of testis and liver were compared between WT and KLHL15^-/^- mice in similar growth conditions; (3) Histopathology: the effects of KLHL15 gene deletion on the structure of liver and testis were observed with HE staining; (4) Liver function: apoptosis of liver tissue was detected with Tunel; liver differential proteins were enriched and analyzed with TMT labeled quantitative proteomics. Results (1) Fertility: compared with WT mice, KLHL15^-/^- mice had decreased total sperm count, increased number of immobile sperm(IM), decreased average curve velocity(VCL)and average path velocity(VAP)(P=0.010, P=0.020). (2) Anatomy: compared with WT mice, KLHL15^-/^- mice had decreased mass of liver and testis(P<0.001, P=0.026), and decreased organ index(P<0.001); (3) Histopathology: the lumen of the convoluted seminiferous tubules of KLHL15^-/^- mice testis became larger, many spermatogenic cells were lost, the epithelial structure became thinner, and the number of sperm decreased; the liver cells were disorderly and the hepatic sinusoids were dilated; (4) Liver function: Tunel results showed more apoptotic cells in the liver of KLHL15^-/^- mice; enrichment analysis showed that deletion of KLHL15 gene affected the composition of mitochondria, various organelle membranes in the liver, reactions involved by peroxisomes and the metabolism of various amino acids. Conclusion Deletion of KLHL15 gene can destroy the structure of testis and reduce the sperm vitality and sperm quality of male mice, leading to reduced fertility. It can also cause structural damage to the liver of mice and affect the metabolic function of the liver.

Key words: KLHL15 gene, Knock out mice, Sperm quality, Liver, Testis

中图分类号:

R588.1

刘笑含,石慧,赵振军. KLHL15基因对雄性小鼠睾丸和肝脏的影响[J]. 山东大学学报 (医学版), 2023, 61(2): 25-30.

LIU Xiaohan, SHI Hui, ZHAO Zhenjun. Effects of KLHL15 gene on the testis and liver of male mice[J]. Journal of Shandong University (Health Sciences), 2023, 61(2): 25-30.

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