67KDa层粘连蛋白在小胶质细胞上的存在及EGCG对其影响的研究

Identification of 67 KDa laminin receptor in microglia and the effects of(-)-epigallocatechin gallate(EGCG)

目的:证明67KDa层粘连蛋白(67LR)在小胶质细胞中的存在,并探讨67LR是否介导了EGCG抑制小胶质细胞的效应。方法:混合培养神经细胞-小胶质细胞,细菌脂多糖(LPS)刺激后分离纯化小胶质细胞,应用免疫荧光细胞双标及Western blot的方法观察小胶质细胞的形态、数量变化,在EGCG干预后观察小胶质细胞形态、数量的变化以及67LR表达量的变化。结果:小胶质细胞上表达一定量的67LR,LPS刺激小胶质细胞后显示其形态变化、数量增多,67LR表达总量增加,给予EGCG干预后小胶质细胞数量减少、胞体变小、67LR的总量减低,接近于静息状态的小胶质细胞。结论:小胶质细胞存在67LR的表达,并可能介导了EGCG抑制小胶质细胞的效应。

Objective：Excessive activation of microglia were closely related to neurodegenerative diseases. Our previous studies showed that（-） -epigallocatechin gallate（EGCG）has a strong inhibitory effects on microglial activation. This study was designed to prove the existence of 67LR in microglial cells, and to explore whether EGCG inhibited microglia through 67LR. Methods： Neuron glia cells were co-cultured for twelve days. Microglial cells were purified by mild digestive method after treated with （lipopolysaccharide （LPS） then observe the changes in microglial morphology and number. Immunofluorescence method was used to observe the presence and expression of 67 KDa laminin receptor （67LR）in microglia. Western blot was applied to detect the change of 67LR amount in microglia after EGCG intervention. Results： 67LR was expressed in microglia. After adding LPS, the number of microglial cells increased and microglial cells transformed into an ameboid shape. LPS could also promote the expression of total 67LR. However, after the administration of EGCG, the number of microglial cells declined and microglial cells recovered to resting state with small body and branches. EGCG could reduce the expression of total 67LR . Conclusion： 67LR is expressed in microglia and can be a potential receptor mediated EGCG inhibiting effects on mi- croglial activation.