Effect of IL-8 and ET-1 on secondary liver injury by hepatic arterial embolization in rabbits

Xin Wang ,Yue Xia Zhong,Li Li Zhang ,Yu Xin Huang ,Oin Sheng Wen , Yan Kui Chu(Department of Gastrointerology,Tangdu Hospital,Fourth Military Medical university,Xian 710038,Shaanxi Province,China)

Abstract：AIM to investigate the mechanism and preventive measures of secondary liver injury by hepatic arterial embolization (HAE) in rabbits.METHODS Animal models of HAE were made with lipiedol ultra-fluid. The rabbits were randomly divided into four groups: shamoperation group (Con); hepatic arterial smbolization group (Amb); hAE plus dexamethasone group {Dex); and HAE plus prostaglandin E group(PGE).Before and after the operation, the plasma interleukine-8(IL-8) and endathelin (ET) and serum ALT, AST and LDHwere measured by ELISA and RIA. changes in liver wereevaluated histopathologically on the sixth day after the operation.RESULTS On the second day after the operation, plasma IL-8 levels of amb group (2456.68μg/L±613.22μg/L)were significantly higher than those in con, Dex and PGE groups (182.83μg/L±49.54μg/L; 189.66μg/L±38.76μg/L;368.21μg/L±178.33μg/L; P＜0.01 ). Plasma IL-8 levels of PGE were higher than those in con(P＜0.05). On the sixth day after the operation, plasma ET levels of Amb, dex and PGE group rose remarkably than Con group (203.66μg/L±35.87μg/L;232.12μg/L±38.24μg/L;210.68μg/L±29.44μg/L; 146.36μg/L±26.22μg/L P＜0.01). On the first day after the operation, serum ALT, AST and LDH levels of Amb(316.67U/L±256.86U/L,444.80U/L±162.48U/L,469.50U/L±168.70U/L ), Dex (342.13U/L±240.54U/L,412.17U/L±252.88U/L, 388.20U/L±67.29U/L ) and PGE ( 259.80U/L±237.66U/L,148.00 u/L±35.90 U/L,552.60 U/L±221.23 U/L) groups were significantly higher then those in Con (31.22U/L±7.74 U/L,16.78 U/L±5.52 U/L,156.60 U/L±36.58 u/L,P＜0.01 ), and they were almost equal to the Con group on he sixth day after the operation. Compared with Amb group, serum ALT and LDH levels dropped remarkably in Dex and PGE group on the second and third days after the operations (P＜0.01). Serum AST levels of PGE group were significantly lower than those in Amb group on the first day after the operation. A lot of neutrophils infiltrated around the embolized liver tissue. Liver cells swelled with piecemeal necrosis in Amb group. But the inflarnrnation around the embolized liver tissue was slighter in Dex and PGE group then those in Amb group. CONCLUSION IL-8 end ET play a role in secondary liver injury by HAE. They are important factors in liver cell injury around the embolized liver tissue. dexamethasone and prostaglandin E can prevent partly the secondary liver injury by HAE through decreasing the production of IL-8 and inhibiting the infiltration of neutrophil. Although the changes of plasma ET levels were not affected by daxarnethesone and prostaglandin E, they can reduce the liver injury by improving the microcirculation of liver and antagonizing the effect of ET。

Keywords：hepatic arterial embolization;rabbit; liver injury; interleukin-8; endothelium

0引言经皮肝动脉栓塞化疗(TACE)是当今世界上治疗不能手术切除的中晚期肝癌的首选方法．无论是肝动脉还是门静脉系统(或同时)桂塞治疗肝癌，其目的是尽可能完全地阻断肿瘤的血供而使其坏死．在临床上，有时TACE继发肝损害常致患者出现腹水、上消化道大出血，甚或肝功能衰竭而使患者死亡，从而使TACE疗效不佳或失败．这种继发性肝损害不仅是栓塞剂过量或操作不当致正常肝组织被栓塞，以及化疗药物的毒性所致，还可能有其他因素参与．白介素8(IL-8)是体内强有效的中性粒细胞趋化因子，在多种肝脏炎症损伤中起重要作用[1]．内皮素(ET)是体内最强的收缩血管物质，是介导多种肝损害的主要递质[1-3]，为此，我们观察了它们在实验性兔肝动脉栓塞继发性肝损伤中的作用．并初步探讨其防治方法，1材料和方法1．1材料TACE动物为日本大耳白兔，32只，雌雄兼有，体质量1．5kg～2．5kg，由第四军医大学实验动物中心提供．超液态碘油法国生产；地塞米松针剂；前列腺素E针剂(商品名勃乐思)，重庆药友制药有限责任公司生产；IL-8试剂盒由第四军医大学免疫学教研室提供；ET试剂盒由北京东亚免疫技术研究所提供，1250mAX线透视机，数字减影成象系统德国产品．全自动生化分析仪，导金8000型，日本产品。1．2方法1．2．1实验分组实验动物随机分为4组，每组8只，单只分笼饲养．假手术组(Con)；仅开腹后，再逐层缝合，不做其他处理．肝动脉栓塞组(Amb)；开腹后，进行肝动脉插管栓塞，然后关腹．肝动脉栓塞+地塞米松治疗组(Dex)：肝动脉栓塞后，每日腹膜腔内注入地塞米松2mg．肝动脉栓塞+前列腺素E治疗组;肝动脉栓塞后，每日腹膜腔内注入前列腺素E15mg．1．2．2肝动脉栓塞动物模型制备20g／L戊巴比妥钠经耳缘静脉麻醉(30mg／kg)，常规消毒，铺巾，上腹正中切口暴露胃，把胃体向右侧翻起，沿腹主动脉寻找胃一肝动脉分枝，沿其主干分离至肝固有动脉，自制穿刺导管插管至肝门部，在x光机透视下注入照影剂显示肝内动脉系统显影，逐注入超液态碘油，数字减影显示形成部分肝动脉栓塞，拔管止血后逐层关腹，送回动物房饲养观察。1.2．3血浆IL-8，ET及血清AST和LDH的测定经兔耳缘静脉于术前，术后1，2，3和6d时各抽血2mL，1mL放入加有100g／LEDTA20μL，抑肽酶500IU的试管内，3500r／min，15min，分离血浆-40°C保存待测．另1mL放人千燥管，室温lh后离心，取上清-40°C保存待测．IL-8用ELISA法测定，ET用RlA法测定．ALT，AST和LDH用自动生化分析仪测定．均按说明书严格操作。1．2．4肝组织学观察术后6d处死动物，取栓塞区及周围肝组织常规固定，石蜡包埋，切片后HE染色，光镜下观察。2结果肝动脉介入部分肝栓塞模型的建立分别用1，2和4mL超液态碘油注入肝动脉，数字减影显示4mL碘油几乎栓塞了大部分肝叶，1mL碘油栓塞区很小，而2mL碘油可造成近1／3的肝右叶栓塞，此实验中均选用2mL碘油栓塞[4]。2，1各组动物肝动脉栓塞前后血浆中IL-8的动态变化肝动脉栓塞后Amb组血浆IL-8水平显著升高(P＜0．01)，2d时达高峰，6d时仍明显高于假手术对照组(Con)(P＜0．01)，Dex组各时间点IL-8水平与Con组无显著差异(P＞0．05)．PGE组仅术后2d时IL-8水平高于Con组(P＜0.05)，但仍明显低于Amb组(P＜0．01)．Con组手术前后各时间点IL-8水平无明显差异(P＞0．05，图1)。图1兔肝动脉栓塞后血浆IL-8含量。2．2各组动物肝动脉栓塞前后血浆ET水产动态变化图2示手术后Con组各时间点血浆ET水平无显著变化(P＞0.05)，至6d时仍继续升高(P＜0．01)，而三组间无显著差异(P＞0．05，图2)。图2兔肝动脉栓塞后ET含量2．3各组动物手术前后血清ALT，AST和LDH水平变化手术前后Con组各时间点ALT，AST和LDH水平无显著变化(P＞0．05)．而Arnb，Dex和PGE三组术后此三项指标均显著升高，1d时达高峰(P＜0．01)．术后6d时各项指标均接近正常．Dex和PGE组术后2d，3d时血清ALT<