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目的探讨抑制素B(INH-B)在不同周龄小鼠睾丸和脑组织中的含量变化及α和β亚基在组织中的分布。方法购买1周龄、4周龄、8周龄和28周龄的雄性C57BL/6小鼠各6只。颈椎脱颈致死后分离小鼠睾丸和脑组织,4℃条件下收集组织匀浆上清,ELISA试剂盒方法检测INH-B含量,蛋白免疫印迹(WB)方法检测INH-Bα和β亚基在组织中的分布。结果 ELISA结果显示,睾丸中的INH-B含量随小鼠周龄增长而上升,在成年小鼠(8周龄)达到峰值,28周龄小鼠水平下降,差异有统计学意义(P<0.05);而脑组织中的INH-B含量在不同周龄小鼠中无显著性差异(P>0.05)。WB结果显示:α亚基单克隆抗体检测识别的蛋白形式在睾丸和脑组织中有所不同,40KD蛋白条带出现在所有小鼠睾丸和脑组织中,39KD和37KD出现在新生1周龄雄性小鼠脑组织中,26KD出现在各周龄小鼠睾丸中;β亚基单克隆抗体检测识别的蛋白主要有17KD、26KD、40KD、55KD以及70KD几种形式,其中26KD蛋白条带仅明显存在于1周龄和4周龄小鼠组织中。结论成熟小鼠脑组织和睾丸中INH-B含量丰富,不同周龄小鼠睾丸和脑组织中α和β亚基的单克隆抗体识别的蛋白条带形式有所不同。
Abstract：Objective: To identify the expression and distribution of inhibin B(INH-B)and itsα&βsubunits of in mice testis and brain.Methods: Twenty-four male C57BL/6mice of one week old,four weeks old,eight weeks and twenty-eight weeks were purchased.The testis and brain tissue were separated after sacrificed,followed by collecting the supernatant of tissue homogenate under the condition of 4℃.The inhibin B levels were measured by ELISA kit.The distribution of INH-B subunits in mice testis and brain were detected by Western blot.Results: ELISA results showed that INH-B content in testis increased with age,and reached the peak at adult mice(eight weeks old),but the INH-B levels were significantly decreased at 28 week old(P<0.05).While the INH-B levels were not significantly different in mice brain among all age groups(P>0.05).The results of Western blot analysis showed that the protein forms identified by α-subunit monoclonal antibody in the testis and brain tissue is different;40KD protein bands appeared in all mice testes and brain tissue;39 KD and 37 KD appeared in the 1 week old male mice brain tissue;26 KDappears in the testes of all aged mice.There were 17 KD,26KD,40 KD,55KD and 70 KD proteins in the detection ofβsubunit monoclonal antibody.The 26 KD protein bands were only presented in 1week old and 4week old mice tissues.Conclusions: The testis and brain of mice are rich in INH-B,especially in adult mice.Different molecular forms of INH-B were detected by antibody forα&βsubunits of inhibin B in the testis and brain of different aged mice.
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基本信息:
中图分类号:R339.21
引用信息:
[1]辛玲,赵海豹,侯丽,等.抑制素B亚基在小鼠睾丸和脑组织中的表达及分布[J].生殖医学杂志,2017,26(06):605-609.
基金信息:
中央级公益科研院所基本科研业务专项(2016GJZ07)
2017-06-15
2017-06-15