褪黑素缓解白消安损伤小鼠睾丸的机理研究

doi:10.11843/j.issn.0366-6964.2024.06.020

摘要/Abstract

摘要：

旨在揭示褪黑素(melatonin，MT)缓解白消安损伤睾丸的机理，提高白消安制备精原干细胞移植受体的效率，并探究其对白消安毒害雄性生精机能的治疗作用。本研究以6~7周ICR雄鼠为模型动物，分为对照组，白消安处理组和白消安+褪黑素处理组，每组8个重复，于注射后第7天取样。通过ELISA检测睾丸组织的炎性细胞因子水平，并对睾丸和附睾进行HE染色。此外，免疫组化或免疫荧光检测血睾屏障(blood-testis barrier，BTB)相关蛋白的表达情况，以分析MT对BTB影响。结果表明，小鼠经白消安注射后，联合MT处理，观察到MT显著缓解了睾丸重和睾体率下降的趋势，并减缓了精子数量下降的速度；睾丸组织HE染色和炎性细胞因子检测发现，MT显著减缓了核转录因子κB(nuclear transportation factor κB，NF-κB)、p-NF-κB、白介素-1β(interleukin-1β，IL-1β)、肿瘤坏死因子-ɑ(tumor necrosis factor-ɑ，TNF-α)及髓过氧化物酶(myeloperoxidase，MPO)水平的上升趋势，从而减少了组织中炎性细胞浸润；免疫组化或免疫荧光检测发现，MT缓解了白消安处理导致的BTB组成蛋白(Occludin、connexin-43、N-cadherin)和细胞骨架蛋白(F-actin和Vimentin)含量的下降。综上，MT通过下调睾丸组织中炎性细胞因子水平，缓解白消安诱发的小鼠睾丸炎症反应；并通过减缓BTB相关蛋白含量的下降，缓解白消安对BTB和支持细胞的损伤，进而缓解小鼠生精机能的损伤。

关键词: 褪黑素, 白消安, 睾丸炎, 血睾屏障, 细胞骨架

Abstract:

The study aimed to reveal the mechanism of melatonin in alleviating testicular damage caused by busulfan, improve the efficiency of busulfan in preparing spermatogonial stem cell transplantation recipients, and explore its therapeutic effect on male spermatogenic function caused by busulfan's toxicity. In this study, ICR male mice at 6-7 weeks were used as model animals and divided into control group, busulfan treatment group, and busulfan+melatonin treatment group. Each group had 8 replicates and samples were taken on the 7th day after injection. The levels of inflammatory cytokines in testicular tissue was detected using ELISA, and perform HE staining on the testes and epididymis. In addition, immunohistochemistry or immunofluorescence were used to detect the expression of blood testis barrier related proteins to analyze the effect of melatonin on BTB. The results showed that after injection of busulfan into mice, combined with melatonin treatment, the trend of decreased testicular weight and testicular body rate was significantly alleviated, and the rate of decrease in sperm count was slowed down; HE staining of testicular tissue and detection of inflammatory cytokines showed that melatonin significantly slowed down the upward trend of nuclear transportation factor κB, p-NF-κB, interleukin-1β, tumor necrosis factor-α and myeloperoxidase, thereby reducing the infiltration of inflammatory cells in the tissue; immunohistochemical or immunofluorescence detection showed that melatonin slowed down the decrease in the levels of blood testis barrier constituent proteins (Occludin, connexin-43, N-cadherin) and cytoskeletal proteins (F-actin and Vimentin) caused by busulfan treatment. Melatonin alleviates the mouse orchitis response induced by busulfan by down regulating the levels of inflammatory cytokines in testicular tissue; and by slowing down the decreasing trend of BTB related proteins content, the damage of busulfan to BTB and sertoli cells is alleviated, thereby alleviating the damage to mouse spermatogenic functionn

Key words: melatonin, busulfan, orchitis, blood testis barrier, cytoskeleton

中图分类号:

S814.1

吴开慧, 张留光, 汪超, 许师源, 刘松奇, 原开敏, 王栋, 庞云渭. 褪黑素缓解白消安损伤小鼠睾丸的机理研究[J]. 畜牧兽医学报, 2024, 55(6): 2486-2497.

Kaihui WU, Liuguang ZHANG, Chao WANG, Shiyuan XU, Songqi LIU, Kaimin YUAN, Dong WANG, Yunwei PANG. Study on the Mechanism of Melatonin Relieving Testicular Damage in Mice Induced Busulfan[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2486-2497.

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