新癀片抗炎镇痛作用机制的蛋白组学研究

目的分析新癀片抗炎镇痛作用机制,通过蛋白组学方法寻找其作用的蛋白靶点。方法大鼠随机分成对照组、模型组、吲哚美辛(2 mg/kg)组以及新癀片23.8、47.5、95.0 mg/kg组。给药组均ig给药10 m L/kg,对照组及模型组ig给予同体积去离子水。1次/d,连续3 d。于末次给药后30 min,除对照组外,在大鼠右后足垫部sc 1%角叉菜胶0.05 m L/只致炎。在致炎前和致炎后1、2 h,记录大鼠抬足潜伏期。剖杀大鼠取肝脏,肝组织蛋白经提取、双向电泳、染色和图谱分析,确定差异表达蛋白,进行蛋白质质谱分析。结果与模型组比较,新癀片23.8、47.5、95 mg/kg组差异表达蛋白个数依次为67、71、94,其中上调个数为10、11、33,下调个数为57、60、61;新癀片95 mg/kg组与吲哚美辛(2 mg/kg)组比较,差异表达蛋白89个,其中上调27个,下调62个。共鉴定出11个差异蛋白质。与模型组比较,新癀片组均可以抑制Shank3、ANXA5、TPI、PSMA2表达,增强PAH、LZTS1表达。结论新癀片可调节的蛋白明显增多,能够抑制多种相关炎症因子和肿瘤因子,增强抗炎因子及抑癌因子的表达,为新癀片"增效"作用机制提供重要理论依据。

Anti-inflammatory and analgesic mechanism of Xinhuang Tablets by proteomics

Objective To analyse anti-inflammatory and analgesic mechanism of Xinhuang Tablets, and to explore drug targeting pathways in protein levels by proteomics technology. Methods SD rats were randomly divided into control group, model group, indomethacin 2 mg/kg group, and Xinhuang Tablets 23.8, 47.5, and 95.0 mg/kg groups. Rats in indomethacin group and Xinhuang Tablets groups were ig administered with the dose of 10 mL/kg, while rats in control group and model group were given the same volume of ionized water, once daily. All rats were treated for 3 d. Except the control group, 1% carrangeenan was sc given into the right footpad of rats 0.05 mL to induce inflammation 30 min after the last administration. Before inflammation and 1 and 2 h after inflammation, foot lift latency was recorded. The animals were sacrificed to obtain livers for proteomics research. The proteins from liver tissue were extracted, two-dimensional electrophoresis, stained, and pattern analysis. Differentially expressed proteins were established, and the identified proteins were determined by the mass spectrometry. Results Compared with the control group, the numbers of differential express proteins in Xinhuang Tablets 23.8, 47.5, and 95.0 mg/kg groups were 67, 71, and 94, respectively. And 10, 11, and 33 proteins in Xinhuang Tablets groups were up-regulated, while 57, 60, and 61 proteins were down-regulated. Compared with the indomethacin group, there were 89 differentially expressed proteins in Xinhuang Tablets groups, including 27 up-regulated proteins and 62 down-regulated proteins. Eleven differentially expressed proteins were identified. Compared with the indomethacin group, Xinhuang Tablets could inhibit the expression of Shank3, ANXA5, TPI, and PSMA2, and enhanced the expression of PAH and LZTS1. Conclusion Xinhuang Tablets can regulate more proteins, and inhibit the expression of inflammatory/tumor-associated factors, enhanced the expression of anti-inflammatory cytokines and tumor suppressor factor, which can provide theoretical basis for impoving efficiency of Xinhuang Tablets mechanism.