原花青素B1对脂多糖介导的炎症反应的抑制作用及其机制探讨

Inhibitory effects and mechanisms of procyanidin B1 on lipopolysaccharide induced inflammatory reaction

目的探索原花青素B1的抗炎活性及其抗炎机制。方法以人急性单核细胞白血病细胞系(THP-1)为模型,设立不含脂多糖(LPS)的磷酸盐缓冲溶液(PBS)处理THP-1细胞为对照组,检测不同浓度原花青素B1(50、100、150、200 mg/L)对THP-1细胞活力的影响。分别应用1 mg/L的LPS(LPS组)、1 mg/L的LPS联合原花青素B1 50 mg/L(处理A组)、1 mg/L的LPS联合原花青素B1 100 mg/L(处理B组)预处理THP-1细胞,收集各组THP-1细胞上清液,应用双抗体夹心亲和素-生物素-过氧化物酶复合物-酶联免疫吸附法测定各组THP-1细胞中TNF-α的释放水平。应用实时定量PCR法测定各组THP-1细胞My D88 m RNA和髓样分化蛋白2(MD-2)m RNA的表达情况。结果不同浓度原花青素的THP-1细胞活力与对照组比较,差异有统计学意义(F=31.35,P<0.001),进一步比较发现,50、100 mg/L原花青素的THP-1细胞活力与对照组比较差异均无统计学意义(P均>0.05),而150、200 mg/L原花青素的THP-1细胞活力均明显低于对照组(P均<0.05)。处理A、B组的TNF-α水平较LPS组均显著减低(P均<0.05),且处理B较处理A组下降更为明显(P<0.05)。对照组、LPS组及处理A组间My D88 m RNA和MD-2m RNA的表达差异均有统计学意义(F=62.500、46.700,P均<0.001),且LPS组的My D88 m RNA和MD-2 m RNA表达均较对照组及处理A组明显升高(P均<0.05),处理A组的My D88 m RNA和MD-2 m RNA表达高于对照组(P均<0.05)。结论原花青素B1可明显抑制LPS介导的炎症反应,负性调节Toll样受体-My D88信号通路,可能是其抑制炎症反应的潜在机制。

Objective To investigate the anti-inflammatory effect of procyanidin B1 and the underlied mechanisms. Methods This study performed in human acute monocytic leukemia cell line （THP-1） cells, excluding lipopolysaccharide （LPS） in phosphate buffer solution （PBS） with THP-1 cells as the control group, the THP-1 cell activity in different concentration of procyanidins B1 （50, 100, 150, 200 mg/L） were detected and compared. THP-1 cells were treated with 1 mg / L LPS （LPS group）, 1 mg / L LPS joint 50 mg/ L procyanidin B1 （group A）, 1 mg/L LPS joint 100mg/L proeyanidin B1 （group B）, respectively. The levels of tumor necrosis factor-alpha （TNF-α） in the supernatants were examined by double-antibody sandwich avidin-biotin-pcroxidase complex enzyme-linked immunosorbant assay. The expression of MyD88 mRNA and myeloid differential protein 2 （MD-2） mRNA were performed by real-time PCR. Results The cell viability of procyanidins B1 in different concentration were significantly different as compared with the control group （F= 31.35, P〈0.001）, and the cell viability of procyanidins B1 with 50, 100 mg/L showed no significant differences as compared with the control group （all P〉 0.05）, the cell viability of procyanidins B1 with 150 and 200 mg/L were much lower than those in the control group （all P 〈 0.05）. The levels of TNF-α in group A and group B were lower than those in the LPS group, and was the lowest in the group B （all P 〈 0.05）. The expression of MyD88 mRNA and MD-2 mRNA in the control group, LPS group and group A all showed significant difference （F= 62.500, 46.700, all P 〈 0.001）, and were much higher in the LPS group than those in the control group and group A, and were higher in the group A than those in the control group （all P〈 0.05）. Conclusion Procyanidin B1 can inhibit LPS-induced inflammatory response through negative modulation Toll-like receptor-MyD88 signaling pathway.