摘要
目的探讨阴囊轻度热应激对睾丸钙/钙调蛋白依赖性蛋白激酶I(p-CaMKII)表达的影响。方法将24只成年ICR雄性小鼠分为4组:对照组、热应激0.5h组热应激2 h组和热应激6h组。热应激组小鼠的处理方式为:将小鼠身体的下1/3部分浸于43℃水浴中15 min,在热应激后0.5.2.6 h取舉丸样品,分别对应为热应激0.5 h组、热应激2 h组和热应激6h组。将对照组小鼠身体下1/3部分浸人22℃水中15 min,6 h后处死并取出睾丸样品。采用脱氧核苷酸末端转移酶(TdT)介导的脱氧三磷酸尿苷(dUTP)缺口末端标记法检测细胞凋亡;用Westerm blot和免疫组织化学技术检测睾丸组织组织磷酸化钙/钙调蛋白依赖性蛋白繳酶II(p-CaMKI)蛋白的表达。结果阴囊热应激能够使睾丸发生明显的病理学损伤,引起小鼠睾丸生殖细胞凋亡。Westemblot和免疫组织化学技术检测结果均表明,热应激组睾丸组织p-CaMK II、p-ERK1/2蛋白的表达水平明显高于对照组。结论单次热应激能激活睾丸CaMKII,CaMKII的激活可能在热诱导睾丸生殖细胞调亡中起重要作用且CaM-℃aMK II和ERK1/2信号通路可能参与其中。
Objective To investigate the efect of a single mild transient scrotal exposure(43 C for 15 min)on the expression of Calcium/calmodulin-dependent protein kinase I intestes.Methods Twenty-four adult ICR male mice were divided into four groups:control group,0.5 h heat stress group,2 h heat stress group and 6 h heat stress group.The mice in heat stress group were treated by immersing the lower third part of the body in a water bath at 43 C for 15 minutes.Testicular samples were taken at 0.5 h,2 h and 6 h after heat stress,corresponding to 0.5 h heat stress group,2 h heat stress group and 6 h heat stress group,respectively.The control mice were im-mersed in a water bath maintained at 22 C and kll at 6 h after heat exposure.Testicular germ cell apoptosis was detemined by terminal dUTP nick-end labelling(TUNEL).The expressions of testicular phosphor-calciun/calm-odulin-dependent protein kinaseⅡ(p-CaMKⅡ)protein were measured using W estern blot and immunostaining.Results A single,acute scrotal bhyperthermia significantly increased testicular germ cell apoptosis,as determined by terminal dUTP nick-end lblling(TUNEL).The expression levels of p-CaMK I and p-ERK1/2 protein in testis of heat stress group were significantly higher than those in control group.Conclusion A single,acute scrotal hy-pertbhermia significantly induces the expression of testicular p-CaMKⅡ,activation of CaMK I may play an impor-tant role in heat-induced apoptosis of testicular germ cells and the CaM-CaMK I and ERK1/2 signaling pathways may be involved.
作者
吴震
朱文祥
刘洪茂
穆柯瀚
姬艳丽
Wu Zhen;Zhu Wenxiang;Liu Hongmao(Dept of Hygiene Inspection and Quarantine,School of Public Health,Anthui Medical University,Hefei 230032)
出处
《安徽医科大学学报》
CAS
北大核心
2019年第12期1938-1942,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:31571557)
2016年高校优秀中青年骨干人才国外访学研修重点项目(编号:gxfxZD2016039)
安徽医科大学“青年拔尖人才支持计划”
