外源性腐胺和尸胺对兔外周血炎症因子的影响

Influence of exogenous putrescine and cadaverine on pro-inflammatory factors in the peripheral blood of rabbits

目的探讨外源性腐胺、尸胺对兔外周血炎症因子的影响。方法取40只普通级成年新西兰兔，按随机数字表法分成生理盐水组、坏死组织匀浆液（简称匀浆液）组、腐胺组、尸胺组，每组10只。生理盐水组兔腹腔注射生理盐水（1mL／kg），匀浆液组兔腹腔注射匀浆液（1mL／kg），腐胺组兔腹腔注射浓度为10g／L的腐胺（1mL／kg），尸胺组兔腹腔注射浓度10g／L的尸胺（1mL／kg）。注射前及注射后2、6、12、24、30、36、48、60h经兔耳中央动脉抽血2mL，采用ELISA法动态观察血清标中TNF-α、IL-1、IL-6的含量变化。对数据进行重复测量方差分析，用Cubic模型曲线对各组炎症因子含量变化进行曲线拟合，并进行Spearman相关分析。结果（1）各组兔血清炎症因子含量变化：①生理盐水组3种炎症因子含量无明显变化。与注射前比较，匀浆液组、腐胺组及尸胺组注射后大部分时相点3种炎症因子含量呈不同程度升高，与生理盐水组比较差异有统计学意义（F值为3．49～13．58，P值均小于0．05）。②腐胺组血清TNF-α、IL-1、IL-6含量分别于注射后2、6、6h明显上升，此趋势与匀浆液组相似，但较尸胺组（均为注射后2h）延迟。③腐胺组血清TNF-α、IL-1、IL-6含量峰值分别为（339±36）、（518±44）、（265．9±33．5）pg／mL，显著低于尸胺组的（476±86）、（539±22）、（309．4±27．1）pg／mL，F值分别为5．11、1．90、5．56，P值均小于0．05。（2）各组兔血消炎症因子含量变化曲线拟合结果：①腐胺组3种炎症因子含量开始上升时间段（注射后3～4h）较尸胺组（注射后2h）延迟，各炎症因子含量达峰值时间段（注射后18～30h）显著晚于尸胺组（注射后12～30h），且峰值持续时间段（注射后18～30h）短于尸胺组（注射后12～30h）。②腐胺组3种炎症因子含量上升时间段和峰值持续时间段均与匀浆液组（分别为注射后3～5、18—30h）较为接近。③相天系数检验分析显示，腐胺组与匀浆液组炎症因子含量变化趋势呈显著相关（rTNF-α=0．933，P〈0．01；rIL-1=0．967，P〈0．01；rIL-6=0．950，P〈0．01），而尸胺组与匀浆液组仅IL-1、IL-6含量变化趋势有一定相关性（rIL-1=0．913，P〈0．01；rIL-6=0．883，P〈0．05）。结论外源性腐胺和尸胺均可引起机体炎症反应，其中腐胺与匀浆液诱导的机体炎症反应较为相似，提示其在坏死组织分解物诱导的炎症反应中可能起重要作用。

Objective To explore the influence of exogenous putrescine and cadaverine on pro-in- flammatory factors in the peripheral blood of rabbits. Methods Forty ordinary aduh New Zealand rabbits were divided into saline, necrotic tissue homogenate （NTH） , putrescine, and cadaverine groups according to the random number table, with 10 rabbits in each group. Saline, NTH, 10 g/L putrescine, and 10 g/L cadaverine were respectively peritoneally injected into rabbits of corresponding group in the amount of 1 mL/kg. The blood sample in the volume of 2 mL was collected from the central artery of rabbit ears before injection and at 2, 6, 12, 24, 30, 36, 48, 60 hours post injection （PIH）. Contents ofTNF-α, IL-1, and IL-6 in the serum were determined with enzyme-linked immunosorbent assay. Data were processed with re- peated measurement data analysis of variance and Spearman correlation analysis, and cubic model curve was applied in curve fitting for the contents of inflammatory factors. Results （ 1 ） The serum contents of TNF-α, IL-1, and IL-6 were increased in NTH, putrescine, and cadaverine groups in different degrees at most post injection time points. There was no significant change in the concentrations of the three pro-inflam- matory factors in saline group, and they were significantly lower than those of the other three groups at most post injection time points （ with F values from 3.49 to 13.58, P values all below 0.05 ）. The serum contents of TNF-α, IL-1, and IL-6 in putrescine group began to increase at PIH 2, 6, and 6, which was similar to the trend of NTH group, but the changes were delayed compared with those of cadaverine group（ all at PIH 2）. The peak values of TNF-α, IL-1, and IL-6 in putrescine group were respectively （339 ±36） , （518 ± 44）, and （265. 9 ± 33. 5） pg/mL, which were significantly lower than those of cadaverine group [（476±86）, （539 ±22）, and （309.4±27. 1） pg/mL, with F values respectively 5. 11, 1.90, and 5.56, P values all below 0.05. （2） The period of time in which contents of TNF-α, IL-1, and IL-6 began to increase （PIH 3-4） and the peaking time of the three pro-inflammatory cytokines （PIH 18-30） in putres- eine group appeared later than those of cadaverine group （PIH 2 and 12-30）. The duration of peaking time of the three pro-inflammatory cytokines in putrescine group was shorter than that of cadaverine group （ PIH 18-30 vs. PIH 12-30）. The increasing period and the duration of peaking time of TNF-α, IL-1, and IL-6 in putrescine group were close to those of NTH group （PIH 3-5 and 18-30）. The correlation coefficient test a- nalysis showed that the trends of changes in contents of three pro-inflammatory cytokines in putreseine group were significantly correlated with those of NTH group （ r TNF-α = 0. 933, P 〈 0.01 ; r IL-1 = 0. 967, P 〈 0.01 ; r IL-6 = 0. 950, P 〈 0.01 ）. The obvious correlation between cadaverine group and NTH group was only found in the contents of IL-1 and IL-6 （ r IL-1 = 0. 913, P 〈 0.01 ; r IL-6 = 0. 883, P 〈 0.05 ）. Conclusions Both exogenous putrescine and cadaverine can cause inflammatory reaction in rabbits. The trend of the in- flammatory reaction induced by putrescine was similar with that by NTH, suggesting that putrescine may play a leading role in the inflammatory reaction induced by necrotic tissue decomposition.