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目的:观察麝香、乳香含药血浆对前列腺抗原刺激诱导的单核细胞炎症因子释放及前列腺细胞或内皮细胞炎症反应相关蛋白表达的影响。方法:采用SD大鼠制备麝香、乳香含药血浆,BALB/c小鼠制备单核细胞与前列腺抗原。单核细胞以梯度浓度(0,2.5%,5%,10%,20%)的麝香、乳香含药血浆预处理1 h后,前列腺抗原诱导活化,培养96 h时ELISA法检测炎症因子释放水平;前列腺细胞RWPE-1与内皮细胞EA.hy926共培养,梯度浓度DP-M&O预处理1 h后,加入经PAgs诱导活化的单核细胞,培养96 h时Western印迹法检测RWPE-1或EA.hy926细胞炎症反应相关蛋白表达水平。结果:麝香、乳香含药血浆随浓度增加,降低TNF-α、IL-1β、IL-6、IL-8及升高IL-10作用逐渐明显,并呈剂量依赖性,20%麝香、乳香含药血浆预处理后,单核细胞释放TNF-α、IL-1β、IL-6、IL-8、IL-10[(70.8±22.3)pg/ml,(277.5±22.6)pg/ml,(232.7±62.7)pg/ml,(227.3±79.2)pg/ml,(640.2±201.2)pg/ml]水平与前列腺抗原组[(277.1±65.5)pg/ml,(630.4±89.7)pg/ml,(994.2±182.3)pg/ml,(769.3±284.1)pg/ml,(271.1±55.8)pg/ml]比较差异有统计学意义(P均<0.01);10%与20%浓度麝香、乳香含药血浆预处理,RWPE-1细胞MCP-1/CCL2(0.56±0.11,0.34±0.08)及EA.hy926细胞VCAM-1(0.52±0.17,0.38±0.12)表达降低,与前列腺抗原组(MCP-1/CCL2:1.12±0.34,VCAM-1:0.94±0.22)比较,差异有统计学意义(P均<0.05)。结论:麝香配伍乳香可通过抑制前列腺上皮细胞MCP-1/CCL2产生,降低血管内皮细胞VCAM-1表达,从而阻断白细胞粘附的关键步骤达到减弱炎症反应。
OBJECTIVE: To observe the effects of musk and frankincense plasma on the release of monocyte inflammatory cytokines and the expression of inflammatory proteins in prostate or endothelial cells induced by prostatic antigen stimulation. Methods: The musk and frankincense drug-containing plasma were prepared from SD rats and monocytes and prostatic antigens were prepared from BALB / c mice. Monocytes were stimulated with prostatic antigens for 1 h after being pretreated with musk and frankincense plasma in gradient concentrations (0, 2.5%, 5%, 10%, 20%). The release of inflammatory cytokines ; RWPE-1 cells were co-cultured with endothelial cell line EA.hy926. After being pretreated with DP-M & O for 1 h, PAgs-induced monocytes were cultured. Western blotting was used to detect RWPE-1 or EA at 96 h. hy926 cell inflammatory response related protein expression levels. Results: The effects of increasing concentrations of musk and frankincense plasma with decreasing concentrations of TNF-α, IL-1β, IL-6, IL-8 and IL-10 were obvious and in a dose-dependent manner with 20% musk and frankincense TNF-α, IL-1β, IL-6, IL-8, IL-10 [(70.8 ± 22.3) pg / ml, (277.5 ± 22.6) pg / ml, (277.1 ± 65.5) pg / ml, (630.4 ± 89.7) pg / ml, (62.3 ± 6.7) pg / ml, 994.2 ± 182.3) pg / ml, (769.3 ± 284.1) pg / ml, (271.1 ± 55.8) pg / ml] were significantly different between the two groups (P <0.01) MCP-1 / CCL2 (0.56 ± 0.11,0.34 ± 0.08) and VCAM-1 (0.52 ± 0.17,0.38 ± 0.12) in RWPE-1 cells were decreased in plasma pretreatment compared with those in prostate-specific antigen group /CCL2:1.12 ± 0.34, VCAM-1: 0.94 ± 0.22), the difference was statistically significant (all P <0.05). Conclusion: Musk compatibility with frankincense can reduce inflammatory reaction by inhibiting the production of MCP-1 / CCL2 in prostate epithelial cells and decreasing the expression of VCAM-1 in vascular endothelial cells, thereby blocking the adhesion of leukocytes.