肿节风注射液对U937细胞共刺激分子表达的影响及其机制

Effect of Sarcandra Glabra Injection on the Expression of Costimulatory Molecules of U937 Cells and Its Mechanism

关键词:  肿节风注射液  U937细胞  CD86  B7-H1  NF-κB

Key words:  Sarcandra Glabra injection  U937cells  CD86  B7-H1  NF-κB

摘要:

目的 探讨肿节风注射液(Sarcandra glabra injection，SGI)对白血病U937细胞共刺激分子表达的影响及其作用机制。方法 用不同浓度(1.83，2.75，3.66，5.49，7.32 g·L^-1)的SGI处理白血病U937细胞48 h后，提取RNA，RT-PCR检测细胞的CD86和B7-H1 mRNA表达；免疫印迹法观察经SGI处理前后CD86、胞浆和核内NF-κB蛋白表达。结果 SGI处理后，B7-H1 mRNA表达降低；在SGI浓度为3.66~7.32 g·L^-1，CD86 mRNA表达升高；CD86蛋白含量随SGI处理浓度增加而升高；胞浆中的NF-κB蛋白含量下降，胞核中含量增加。结论 SGI在一定浓度下能诱导U937白血病细胞CD86表达，下调B7-H1的表达，其机制可能与核内NF-κB含量增加有关。

Abstract:

OBJECTIVE To explore the effect of Sarcandra glabra injection(SGI) on the expression of costimulatory molecules of U937 cells and its potential mechanism. METHODS The total RNA was extracted when monocytic leukemia U937 cells were treated with different concentration(1.83, 2.75, 3.66, 5.49, 7.32 g·L^-1) of SGI for 48 h. The expressions of costimulatory molecules CD86 and B7-H1 mRNA were detected by RT-PCR. The level of CD86 and NF-κB protein in cytoplasm or intranuclear was measured by Western blotting. RESULTS When treated with SGI for 48 h, the expression of B7-H1 mRNA decreased, the concentration of CD86 protein increased as the concentration of SGI increased, the expression of CD86 mRNA increased when the concentration of SGI were 3.66-7.32 g·L^-1, the concentration of intracytoplasmic NF-κB protein decreased, the intranuclear NF-κB protein increased. CONCLUSION SGI could enhance the expression of CD86 of U937 cells, and reduce the expression of B7-H1. Its mechanisms might be related to the increase of content of intranuclear NF-κB protein.