论文部分内容阅读
目的:探讨八肽胆囊收缩素(CCK-8)对内毒素血症小鼠腹腔巨噬细胞B7.1和B7.2表达及其协同刺激活性的影响。方法:将BALB/c小鼠随机分组(n=4),分别腹腔注射生理盐水(0.2-0.3 mL/mouse),LPS(100μg/mouse)和/或CCK-8(5 nmol/mouse)及CR1409(100μg/mouse)、CR2945(100μg/mouse)。12 h后收集并纯化腹腔巨噬细胞。采用流式细胞术分析细胞表面B7.1和B7.2含量的变化。用免疫磁珠从小鼠脾细胞分离CD4+T细胞,按4∶1数量比与上述处理的腹腔巨噬细胞共同体外培养,同时加入ConA5 mg/L,采用[3H]掺入法测定CD4+T细胞增殖,反映巨噬细胞的协同刺激活性。结果:整体应用CCK-8作用小鼠腹腔巨噬细胞,与对照组相比,CCK-8可上调静息小鼠腹腔巨噬细胞B7.2的表达,而对B7.1的表达则无影响;并且CCK-8使CD4+T细胞[3H]-TdR的掺入率升高,即促进其增殖,增强巨噬细胞的协同刺激活性。CCK-8降低LPS活化的内毒素血症小鼠腹腔巨噬细胞B7.1和B7.2的表达并且降低CD4+T细胞的[3H]-TdR掺入率,即抑制其增殖,抑制其协同刺激活性。CR1409及CR2945均能逆转CCK-8的上述作用,且CR1409的作用较CR2945更明显。结论:CCK-8通过上调巨噬细胞B7.2表达而增强其协同刺激活性;并且降低LPS活化的内毒素血症小鼠腹腔巨噬细胞B7.1和B7.2的表达,抑制其协同刺激活性。该作用由CCK1R及CCK2R共同介导,其中CCK1R起主要介导作用。
Objective: To investigate the effect of cholecystokinin octapeptide (CCK-8) on the expression of B7.1 and B7.2 peritoneal macrophages and their synergistic stimulatory activity in endotoxemic mice. Methods: BALB / c mice were randomly divided into four groups: normal saline (0.2-0.3 mL / mouse), LPS (100 μg / mouse) and / or CCK-8 (100 μg / mouse), CR2945 (100 μg / mouse). After 12 h, peritoneal macrophages were collected and purified. The changes of cell surface B7.1 and B7.2 contents were analyzed by flow cytometry. CD4 + T cells were isolated from mouse spleen cells by immunomagnetic beads and cultured in vitro in the ratio of 4: 1 with the above-treated peritoneal macrophages. At the same time, ConA5 mg / L was added and CD4 + T Cell proliferation, reflecting the synergistic stimulatory activity of macrophages. Results: Compared with the control group, CCK-8 could up-regulate the expression of B7.2 in peritoneal macrophages in CCK-8 mice, but had no effect on the expression of B7.1 ; And CCK-8 increased the incorporation rate of [3H] -TdR in CD4 + T cells, which promoted the proliferation and enhanced the co-stimulatory activity of macrophages. CCK-8 decreases the [3H] -TdR incorporation, decreases the proliferation of CD4 + T cells and inhibits the synergistic effect of CCK-8 on the expression of B7.1 and B7.2 in peritoneal macrophages of LPS-activated endotoxemic mice Stimulate activity. Both CR1409 and CR2945 reversed the above effects of CCK-8, and the effect of CR1409 was more pronounced than that of CR2945. CONCLUSION: CCK-8 enhances its synergistic stimulatory activity by up-regulating the expression of B7.2 in macrophages, and decreases the expression of B7.1 and B7.2 in peritoneal macrophages in LPS-activated endotoxemic mice and inhibits its synergistic stimulation active. This effect is mediated by both CCK1R and CCK2R, of which CCK1R plays a major mediating role.