DBTC诱导SD大鼠慢性胰腺炎及血清AMS、TNF-α含量测定

DBTC Induced SD Rat Model of Chronic Pancreatitis and Determination of Serum AMS and TNF-α Levels

目的应用二氯二丁基酯(DBTC)诱导SD大鼠慢性胰腺炎(CP),研究CP发生发展过程中病理学变化及血清淀粉酶(AMS)、肿瘤坏死因子(TNF-α)含量变化。方法实验组鼠尾静脉注入0.8mg/kg/DBTC80%乙醇液,对照组注入等量80%乙醇液,实验组鼠分别于第3、14、28d和56d处死,对照组鼠于第56d处死,均行大体肉眼观察及胰腺、肝、肺、肾病理组织学观察及评分。血清AMS含量由全自动生化分析仪测定,血清TNF-α含量测定为ELISA法。结果3d组胰腺大体形态及组织学形态基本正常,其他各组胰腺大体形态改变包括不同程度的肿胀、出血、坏死及与周围组织有粘连等;镜下组织学观察发现:14d组(8只)1只轻度CP;28d组(10只),5只轻度CP,1只中度CP和1只重度CP;56d组(12只)4只轻度CP,3只中度CP和4只重度CP;28d组和56d组病变程度评分、纤维化程度评分及CP发生率明显高于14d组(P<0.05或P<0.01),28d组病变程度评分和纤维化程度评分明显低于56d组(P<0.05)。实验组中部分肝和肺大体形态表现为不同程度出血、坏死及脓肿形成,肾无明显肉眼形态改变;镜下表现与大体形态表现基本一致。对照组胰腺、肝、肺和肾大体形态及组织学形态表现均正常。各实验组血清AMS、TNF-α含量均明显高于对照组(P<0.01),28d组明显低于14d组和56d组(P<0.05或P<0.01),56d组和14d组之间无明显差异(P>0.05)。结论DBTC一次性尾静脉注入能成功地建立SD大鼠CP模型,具体操作简单、时间短、发病率高及费用低廉等优点,但对肝和肺等主要脏器有一定非致死性毒副作用。该模型较符合人类CP病理形态学特征及血清AMS、TNF-α含量改变,TNF-α在CP发生发展中可能起重要作用。

Objective To establish a model of chronic pancreatitis （CP） in Sprague- Dawely （SD） rats induced by single tail vein injection of dibutyltin dichloride （DBTC）, and to observe the pathology and serum AMS, TNF a alterations during the development of CP. Metheds SD rats in the experimental group were treated by single tail vein injection with 0.8mg/kg DBTC in 80% ethanol, and the same volume of 80% ethanol was injected into the control group rats. The rats of experimental group were sacrificed at 3d, 14d, 28d and 56d respectively after injection of DBTC and the control group rats were sacrificed at 56d. The pancreas and major organs including liver, lungs and kidneys were inspected macro - and micro - scopicaliy, the levels of serum AMS were detected by autobiochemical assay apparatus, and the levels of serum TNF - a were determined by ELISA. Results No macro- and micro- scopic pathology of pancreas, liver and lung was found in the 3d group, but the macroscopic pathology including interstitial edema, hemorrhage, abscess and conglutination in the adjacent tissues showed in the other experimental groups. Microscopic histology revealed （1/8） mild CP in the 14d group, 7/10 cases including 5 mild CP, 1 middle and 1 severe CP in the 28d group, but in the 12 rats of the 56d group, only 1 case was normal, 4 cases with mild CP, 3 moderate and 4 severe CP. The pathological evaluation, fibrosis degree and prevalence of CP were increased significantly in the 28d and the 56d groups than that in the 14d group （P〈0.05 or P〈0.01）. The changes of liver, lung and kidney were in accordance with the general pathology. No pathological change of pancreas, liver, lung and kidney was observed in the control group. The levels of serum AMS and TNF - α of experimental group rats were significantly higher than those in the control group rats （P〈0.01）, and the levels of serum AMS and TNF-α were significantly lower in the 28d group than those in the 14d and the 56d groups （P〈0.05 or P〈0.01）. Conclusions The single tail vein injection of DBTC successfully induces CP of SD rats with a simple method, spending less time, high prevalence and low cost. But to a certain extent, DBTC has the toxic side - effect to some major organs, such as liver and lung. The model of SD rat induced by DBTC is similar with human CP in pathological characters and levels of serum AMS and TNF - α, and serum TNF - αmay play an important role in the development of CP.