双氢睾酮和褪黑素在牦牛毛囊细胞中的互作效应研究  

Study on the Interaction Between Dihydrotestosterone and Melatonin of Hair Follicle Cells in Yak

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作  者:张哓兰 赵志东[1] 石斌刚 胡江[1] ZHANG Xiaolan;ZHAO Zhidong;SHI Bingang;HU Jiang(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China)

机构地区:[1]甘肃农业大学动物科学技术学院,兰州730070

出  处:《中国畜牧兽医》2024年第12期5198-5208,共11页China Animal Husbandry & Veterinary Medicine

基  金:甘肃省青年科技基金(23JRRA1435);甘肃农业大学学科团队项目(GAU-XKTD-2022-22)。

摘  要:【目的】揭示双氢睾酮(dihydrotestosterone,DHT)和褪黑素(melatonin,MLT)在牦牛毛囊细胞中的互作效应,为阐明这两种激素在毛囊周期发育中的调控机制及其作为重要的信号分子在牦牛毛发性状选育中的应用提供依据。【方法】以牦牛真皮毛乳头细胞(dermal papilla cells,DPCs)和表皮毛母质细胞(hair matrix cells,HMCs)为研究对象,利用噻唑蓝(MTT)比色法测定不同浓度DHT(0、0.01、0.1、1、10、20、40、80μmol/L)和MLT(0、250、500、600、750、1000、1500 pg/mL)对牦牛DPCs和HMCs增殖的影响;选择最佳浓度DHT和MLT联合处理HMCs,采用MTT法检测两种激素在牦牛HMCs中的互作效应,利用EdU进一步验证试验结果。利用实时荧光定量PCR检测DHT和MLT联合处理后细胞增殖及毛囊发育相关基因的表达情况,分析DHT和MLT可能的作用机制。【结果】与对照组(0μmol/L)相比,10~40μmol/L DHT可显著或极显著促进DPCs增殖(P<0.05;P<0.01),40~80μmol/L DHT可极显著促进HMCs增殖(P<0.01),其中40μmol/L DHT为促进DPCs和HMCs增殖的最佳浓度,此外,低浓度DHT(1μmol/L)可极显著抑制HMCs增殖(P<0.01)。与对照组(0 pg/mL)相比,500 pg/mL MLT对DPCs的促增殖作用最为明显(P<0.01),而不同浓度MLT对HMCs增殖均无明显作用(P>0.05)。使用40μmol/L DHT和500 pg/mL MLT联合处理HMCs 72 h,与DHT组相比,DHT+MLT联合处理对HMCs的促增殖作用极显著降低(P<0.01)。实时荧光定量PCR结果显示,与对照组相比,DHT处理HMCs后极显著或显著改变了与毛囊发育相关基因LEF 1、SFRP 1、TGFB 2和BCL 2的表达量(P<0.01;P<0.05),提示DHT可能通过影响这些基因的表达参与调控毛囊的发育过程。【结论】牦牛DPCs和HMCs对DHT和MLT作用的敏感性不同,一定浓度的DHT均可显著促进DPCs和HMCs的增殖,低浓度DHT具有抑制HMCs增殖的趋势。500 pg/mL MLT是促进DPCs增殖的最佳浓度,不同浓度MLT对HMCs增殖均无明显作用。在HMCs中,MLT具有抗DHT作用的功能。【Objective】This study was aimed to reveal the interaction effect between dihydrotestosterone(DHT)and melatonin(MLT)of hair follicle cells in yak,so as to provide a basis for elucidating the regulatory mechanism of the two hormones in the development of hair follicle cycle and their application in hair trait selection of yak.【Method】Using the dermal papilla cells(DPCs)and hair matrix cells(HMCs)of yak as research subjects,the effects of different concentrations of DHT(0,0.01,0.1,1,10,20,40 and 80μmol/L)and MLT(0,250,500,600,750,1000 and 1500 pg/mL)on the proliferation of DPCs and HMCs in yak were determined by MTT assay.The concentrations of DHT and MLT that had the most significant effect on hair follicle cells were selected for combined treatment of HMCs and their interaction effect of the two hormones in cell proliferation was detected using MTT method.The test result was further verified by EdU method.The expression of several genes related to cell proliferation and hair follicle development were detected by Real-time quantitative PCR after combined treatment of DHT and MLT,and analyze the underlying mechanisms.【Result】Compared with control group(0μmol/L),DHT concentrations ranging from 10 to 40μmol/L could significantly or extremely significantly promote the proliferation of DPCs(P<0.05 or P<0.01),and the concentration of DHT at 40-80μmol/L could extremely significantly promote the proliferation of HMCs(P<0.01).40μmol/L DHT was the optimal concentration to promote the proliferation of DPCs and HMCs,while low concentration of DHT(1μmol/L)could extremely significantly inhibit the proliferation of HMCs(P<0.01).Compared with control group(0 pg/mL),500 pg/mL MLT had the most significant proliferative effect on DPCs(P<0.01),while all concentrations of MLT had no significant effect on the proliferation of HMCs(P>0.05).When HMCs were combining treated with 40μmol/L DHT and 500 pg/mL MLT for 72 h,compared with DHT group,DHT+MLT extremely significantly reduced the proliferative effect of HMCs(P<0.01)

关 键 词:牦牛 毛乳头细胞(DPCs) 毛母质细胞(HMCs) 双氢睾酮(DHT) 褪黑素(MLT) 

分 类 号:S852.2[农业科学—基础兽医学]

 

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