人外周血淋巴细胞和粒细胞褪黑素受体亚型mt1 的检测

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370bp，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，提示褪黑素在生理及病理情况下可能对免疫系统有调节作用。

Detection of melatonin receptor subtype mt1 in human peripheral blood lymphocytes and granulocytes

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.