姜黄素类似物A13的抗炎机制及其在SMIR术后持续性疼痛中的作用研究

Curcumin analogue A13 anti-inflammatory mechanism and its role in persistent pain after SMIR

目的针对本课题组活性好的姜黄素类似物A13,深入研究其抗炎机制及镇痛作用。方法建立大鼠SMIR术后持续性疼痛动物模型,进行行为学测定,测定术前及术后1、3、7、12、22、32 d机械性缩足反射阈值(MWT)和热缩足潜伏期(TWL)。提取原代腹腔巨噬细胞采用细胞NF-κB p65核转移试剂盒进行细胞免疫荧光实验,观察脂多糖(LPS)诱导下A13作用后NF-κB P65核转移情况。结果SMIR模型组术后1、3、7、12、22 d MWT均显著低于术前,差异有统计学意义(P<0.05),术后12 d(7.2±0.4)下降最为明显(P<0.05),术后32 d(13.6±0.7)接近于术前水平。与SMIR模型组比较,SMIR+A13给药组术后1、3、7、12、22 d抑制MWT下降,MWT均显著高于SMIR模型组同等时间点,差异有统计学意义(P<0.05),更接近于空白对照组。与Control组比较,SMIR+A13给药组术后3、7 d两天显著低于Control组同等时间点,差异有统计学意义(P<0.05)。三组大鼠的TWL均无明显改变,无显著性差异(P>0.05)。A13能够抑制LPS诱导下从细胞质到细胞核的p65核转移。结论姜黄素类似物A13在SMIR术后持续性疼痛中对机械性痛觉具有明显镇痛作用,但对热痛觉无明显作用,其通过抑制NF-κB通路达到抗炎效果。

Objective To investigate the anti-inflammatory mechanism and analgesic effect of curcumin analog A13 with good activity.Methods An animal model of persistent pain after SMIR operation was established in rats,and behavioral measurements were performed to determine the mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)before and after 1 d,3 d,7 d,12 d,22 d,and 32 d of treatment.The primary peritoneal macrophages were extracted.The cell NF-κB p65 nuclear transfer kit was used to perform cellular immunofluorescence experiments.The nuclear transfer of NF-κB P65 after lipopolysaccharide(LPS)-induced A13 was observed.Results In the SMIR model group,the MWT at 1,3,7,12,and 22 days after operation was significantly lower than that before operation,and the difference was statistically significant(P<0.05).The MWT(7.2±0.4)was decreased most obviously at the 12th day after operation(P<0.05),and the MWT(13.6±0.7)of the 32nd day was close to the preoperative level.Compared with the SMIR model group,the SMIR+A13 administration group inhibited the decline of MWT at 1,3,7,12 and 22 days after operation.The MWT in the SMIR+A13 administration group was significantly higher than that of the SMIR model group at the same time point,and the difference was statistically significant(P<0.05),which was closer to the blank control group.Compared with the control group,the MWT of the SMIR+A13 administration group was significantly lower than that in the control group at the same time point on the 3rd and 7th days after the operation,and the difference was statistically significant(P<0.05).There was no significant change in TWL of the three groups,and the results were not significantly different(P>0.05).A13 can inhibit p65 nuclear transfer from cytoplasm to nucleus induced by LPS.Conclusion Curcumin analogue A13 has obvious analgesic effect on mechanical pain in persistent pain after SMIR operation,but has no obvious effect on thermal pain,which can achieve anti-inflammatory effect by inhibiting NF-κB pathway.