槲皮素和姜黄素对脓毒症大鼠炎症因子的影响

Effects of quercetin and curcumin on cytokine in septic rats

目的探讨槲皮素和姜黄素预处理对脓毒症大鼠肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）及核转录因子一KB（NF-κB）的影响。方法将120只Wistar大鼠按随机数字表法分为假手术组（A组）、模型组（B组）、槲皮素预处理组（C1组100mg／kg、C2组50mg／kg）、姜黄素预处理组（D1组100mg／kg、D2组50mg／kg）、槲皮素＋姜黄素预处理组（E1组槲皮素50mg／kg＋姜黄素100mg／kg，E2组槲皮素50mg／kg＋姜黄素50mg／kg，E3组槲皮素100mg／kg＋姜黄素50mg／kg，E4组槲皮素100mg／kg＋姜黄素100mg／kg）。采用盲肠结扎穿孔术（CLP）复制脓毒症模型；A组仅开腹分离盲肠而不结扎。各组于制模前灌胃相应药物，A组和B组仅灌胃0．1％二甲基亚砜（DMsO），均每日1次，连用3d。各组取6只大鼠记录制模后脓毒症严重程度评分及生存时间；余大鼠于制模后24h处死，用酶联免疫吸附试验（ELISA）测定血清TNF-α、IL-6水平，荧光定量聚合酶链反应（PCR）测定肝组织核转录因子-κB（NF-κB）mRNA表达，并分析脓毒症严重程度评分、NF-κB与TNF-α、IL-6的相关性。结果①B组术后24h脓毒症严重程度评分（分）明显高于A组（P〈0．05），半数生存时间较C、D、E组明显缩短（P〈0．05）；各药物预处理组术后脓毒症严重程度评分均较B组明显下降（均P〈0．05），其中E4组（13．83±1．33）下降更显著。②B组TNF-α、IL-6、NF-κBmRNA表达均较A组明显升高（均P〈0．05）；与B组比较，药物预处理组能明显降低TNF-α、IL-6的浓度和NF_KBmRNA的表达水平，且各组组内比较差异有统计学意义（均P〈0．05），其中以E4组下降更显著。③脓毒症严重程度评分与血清TNF-α、IL-6水平均呈正相关（r值分别为0．852、0．878，均P〈0．01）；NF-κBmRNA与血清TNF-α、IL-6水平呈正相关（，值分别为0．852、0．848，均P〈0．01）。结论槲皮素和姜黄素预处理能明显抑制脓毒症时NF-κB的表达和炎症反应，随槲皮素及姜黄素剂量增加作用更为显著；且二药合用较单药效果更佳。

Objective To study the effects of quercetin and curcumin pre-treatment on tumor necrosis factor-α （TNF-α）, intedeukin-6 （IL-6） and nuclear factor-κB （NF-κB） in septic rats. Methods According to the random digits table, 120 Wistar rats were divided into following groups （each n = 12 rats） ： sham-operated group （group A）, the model group （group B）, quercetin pre-treatment groups （group C1 and C2, the dosage of quercetin 100 mg/kg and 50 mg/kg respectively）, cureumin pre-treatment groups （group D1 and D2, the dosage of curcumin 100 mg/kg and 50 mg/kg respectively）, quercetin plus curcumin groups （group E1 ： 50 mg/kg of quercetin plus 100 mg/kg of curcumin, group E2 ： 50 mg/kg of quercctin plus 50 mg/kg of curcumin, group E3 ： 100 mg/kg of quercetin plus 50 mg/kg of curcumin, group E4 ： 100 mg/kg of quereetin plus 100 mg/kg of curcumin）. The rat model of sepsis was reproduced by cecal ligation and perforation （CLP）. In group A, the cecum was isolated without ligation. Before the establishment of rat model, all the rats were administered with quercetin and curcumin intra-gastrically with the different dosages as mentioned above in different groups. In group A and group B, the rats without medical pre-treatment received 0.1% dimethyl sulfoxide （DMSO）via intragastric administration, once every day for 3 days. After the establishment of model, the sepsis score and the survival time of 6 rats in each group were observed. The others in each group were sacrificed at 24 hours after operation and their sepsis scores were recorded. The levels of TNF-α and IL-6 were detected by enzyme-linked immunosorbent assay （ELISA）and the mRNA expression of NF-κB in liver tissue was detected by fluorescent quantitative real-time polymerase chain reaction （PCR）. The correlations of sepsis severity score, NF-κB and TNF-α, IL-6 were analyzed. Results ① The sepsis score in group B was higher than that in group A after operation for 24 hours （P〈0.05）. Compared with groups C, D and E, the semi-survival time was significantly shortened in group B （P〈0.05）. Compared with group B, the sepsis scores of medicine pre-treated groups were significantly decreased, especially obvious in group E4 （13.834±1.33, P〈0.05）. ②In group B, the levels of TNF-oL, IL-6 and NF-κB mRNA expression were higher than those in group A （all P〈 0.05）. And the medicine pre-treated groups showed an obvious descent in these indexes （all P〈0.05）, especially marked in group E4. ③ The correlations between sepsis score and TNF-α, IL-6 levels were positive （r=0.852, 0.878, both P〈0.01）. The correlations between NF-κB mRNA expression and TNF-α, IL-6 levels were also positive （r = 0.852, 0.848, both P〈0.01）. Conclusions The results demonstrate that quercetin and curcumin can significantly inhibit the expression of NF -κB and the inflammatory reaction in sepsis. With the increase of quercetin or curcumin in dosage, the effect is more significant, and the therapeutic effect of quercetin and curcumin in combinationis better than that of either one of them alone.