人外周血淋巴细胞和粒细胞褪黑素受体亚型mt_1的检测  

Detection of melatonin receptor subtype mt_1 in hum an peripheral blood lymphocytes and granulocytes

作　　者：陈向芳[1] 刘志民[1] 邓安梅[2] 仲人前[2] 赵瑛[3] 林炜栋[4] 彭树勋[5] 

机构地区：[1]上海第二军医大学附属长征医院内分泌科,200003 [2]上海第二军医大学附属长征医院检验科,200003 [3]上海第二军医大学附属长征医院神经内科,200003 [4]上海第二医科大学附属瑞金医院烧伤中心 [5]香港大学医学院生理系

出　　处：《中华内分泌代谢杂志》2003年第5期411-413,I004,共4页Chinese Journal of Endocrinology and Metabolism

基　　金：香港裘槎基金资助(396 6 30 0 8)

摘　　要：目的　检测人外周血淋巴细胞和粒细胞的褪黑素受体 (MR)亚型mt1和其细胞内定位。方法　分离出健康成年人外周血淋巴细胞和粒细胞 ,应用RT PCR技术分别检测其亚型mt1的mRNA ,阳性产物用自动测序仪测序 ;同时将所分离的淋巴细胞和粒细胞制成细胞涂片 ,应用免疫组化染色检测mt1在淋巴细胞和粒细胞上的分布。结果　RT PCR产物电泳显示 :人外周血淋巴细胞和粒细胞均存在mt1阳性条带 ,片段长度约 3 70bp ,测序结果显示扩增产物与GenBank的人mt1受体亚型的基因序列相吻合 ;免疫组化结果显示 :人外周血淋巴细胞和粒细胞均存在mt1受体蛋白 ,分布于细胞膜、细胞浆和细胞核 ,以细胞膜和细胞浆为主。结论　人外周血淋巴细胞和粒细胞中均存在MR的亚型mt1,外周血淋巴细胞和粒细胞为褪黑素作用的外周靶器官 。Objective To as sa y the melatonin receptor (MR) subtype mt 1 and its localization in the human pe ripheral blood lymphocytes and granulocytes. Methods Lymphocytes and granulocytes of peripheral blood from healthy volunteers w ere isolated by gradient centrifugation. RT-PCR was applied to examine mt 1 mR NA of the lymphocytes and granulocytes, and target products were sequenced. The immunohistochemistry Envision method was applied to oberserve the distribution o f mt 1 in lymphocytes and granulocytes. Results RT-PCR analysis identified the existence of mt 1 mRNA in the cells investiga ted. Sequencing of RT-PCR product showed that the DNA sequence of product coinc ided with the sequence from Genbank. The mt 1 were observed in human peripheral blood lymphocytes and granulocytes by immunohistochemistry, and positive granul es of mt 1 were scattered primarily in cellular membrane and cytoplasm and rare ly in nucleus. Conclusion MR subtype mt 1 is p resent in human lymphocytes and granulocytes from peripheral blood. It suggests that melatonin may play a role in regulative effects of immune system.

关 键 词：人外周血淋巴细胞 粒细胞褪黑素受体亚型mt1 检测 免疫组化染色 粒细胞 

分 类 号：R446.9[医药卫生—诊断学] R446.6[医药卫生—临床医学]