早期间断腹腔灌洗联合抗生素对重症急性胰腺炎大鼠的影响

目的 通过动物实验初步评估早期间断腹腔灌洗(PL)联合抗生素对重症急性胰腺炎(severe acute pancreatitis,SAP)治疗的影响.方法 成年 Wistar大鼠56只,雌雄各半,随机分组:假手术(SO)对照组、SAP无抗生素+未置管组(SNANT)、SAP无抗生素+置管组(SNAT)、SAP抗生素+置管组(SAT)、SAP抗生素+未置管组(SANT)、SAP-PL抗生素组(APL)和SAP-PL无抗生素组(NAPL);每组8只.采用胆胰管逆行性缓慢注射5%牛黄胆酸钠溶液(0.02 ml/100 g)建立大鼠SNANT和SANT模型;在SNANT模型的基础上于右下腹部放置腹腔引流管建立大鼠重症急性胰腺炎SNAT、SAT、APL和NAPL模型,引流管固定于鼠尾部.PL组在成功造模后12 h于麻醉状态下行腹腔灌洗,每12 h 1次,共4次.应用抗生素组则于造模之后12 h或灌洗结束时行腹腔内注射抗生素[左氧氟沙星注射液40 mg/(kg·d)+甲硝唑注射液90 mg/(kg·d)],每12 h 1次,共4次.用药和(或)灌洗完成12 h后收集标本.用酶联免疫吸附测定(ELISA)法检测血清中细胞因子肿瘤坏死因子(TNF-α)、白介素-6(IL-6)、白介素-8(IL-8)、内毒素即脂多糖(LPS)的含量变化,HE染色光镜下观察胰腺组织病理形态学改变并评分.结果 ①与SO组相比,SAP组和PL组血中LPS、TNF-α、IL-6、IL-8水平均显著升高(P＜0.05);与SNANT组相比,SNAT组和NAPL组血中LPS、TNF-α、IL-6、IL-8水平无显著差异(P＞0.05);与SANT组相比,SAT组和APL组血中LPS、TNF-α、IL-6、IL-8水平无显著差异(P＞0.05);与SNANT组、SNAT组和NAPL组相比,SANT组、SAT组和APL组血中LPS、TNF-α、IL-6、IL-8水平均显著降低(P＜0.05);与NAPL组相比,APL组血中LPS、TNF-α、IL-6、IL-8水平均显著降低(P＜0.05).②胰腺病理检查:SAP各组胰腺组织小叶间隙增大、粒细胞浸润、脂肪坏死、实质出血、小叶坏死或缺失;PL各组胰腺组织小叶间隙较SAP组明显增大、粒细胞浸润、脂肪坏死、实质出血、小叶坏死或缺失;SO组仅见散发胰腺小叶间区域水肿.SAP各组、PL各组胰腺病理学评分显著高于SO组(P＜0.05);PL各组胰腺组织病理学总评分与SAP各组胰腺组织病理学总评分相比无统计差异(P＞0.05).结论 重症急性胰腺炎早期间断腹腔灌洗联合抗生素治疗可显著降低SAP大鼠血中LPS、TNF-α、IL-6、IL-8水平(P＜0.05),从而可能有助于降低SAP的全身损害,达到促进SAP好转和改善预后的目的.

Study on the therapeutic effect of interrupted peritoneal lavage combined with antibiotics on severe acute pancreatitis in rats

Objective To evaluate the therapeutic effect of interrupted peritoneal lavage(PL) combined with antibiotics on the severe acute pancreatitis in rats.Methods Fifty-six adult Wistar rats(280 g-350 g) were randomly divided into sham operation(SO) group,SAP groups of non-antibiotics+non-drainage tube(SNANT) group,of non-antibiotics+drainage tube(SNAT) group,of antibiotics+drainage tube(SAT) group,of antibiotics + non-drainage tube(SANT),of SAP+PL groups included antibiotic+peritoneal lavage(APL) group,and of non-antibiotics+peritoneal lavage(NAPL) group,with 8 rats in each group.SNANT and SANT models were produced by slow retrograde infusion of 5% sodium taurocholate solution(0.02ml/100g) into the common biliopancreatic duct in Wistar rats.In the SNAT、SAT、APL and NAPL groups,abdominal cavity drainage tube with side holes was introduced into the abdominal cavity at the right lower quadrant of the abdomen based on the SNANT models,and was tied to the rat tail.Peritoneal lavage was performed with normal saline at ambient temperature and narcosis after 12 hours of successful induction of animal models,once per 12 hours and four times in total.The intraperitoneal injection of antibiotics(Left oxygen Buddha injection 40 mg/kg+Metronidazole Injection 90 mg/kg) was performed once in every 12 hours after successful induction of animal models in SANT and SAT groups or next to the termination of peritoneal lavage in APL group.Blood sample and pancreatic tissue were obtained after 12 hours of injection of antibiotics and peritoneal lavage.The blood levels of tumor necrosis factor-α(TNF-α),Interleukin-6(IL-6),Interleukin-8(IL-8),lipopolysaccharide(LPS) were measured by enzyme-linked immunosorbent assay(ELISA).The changes of pathological morphology of pancreatic tissue were evaluated and scored under light microscope after staining with hematoxylineosin-stain.Results(1) Compared with the SO group,the blood levels of TNF-α、IL-6、IL-8、LPS in SAP groups and SAP+PL groups were increased remarkably(P<0.05);Compared with the SNANT group,there were no significantly changes(P>0.05) in the SNAT group and the NAPL group;Compared with the SANT group,there were no significantly changes(P>0.05) in the SAT group and the APL group in the blood levels of LPS、TNF-α、IL-6、IL-8;Compared to the groups without antibiotics including SNANT group,SNAT group and NAPL group,the blood levels of LPS、TNF-α、IL-6、IL-8 in the groups with antibiotics including SANT group,SAT group and APL group were decreased remarkably(P<0.05);Compared with the NAPL group,the blood levels of LPS、TNF-α、IL-6、IL-8 in the APL groups were decreased remarkably(P<0.05).(2) The histological examination of pancreatic tissue showed that: the enlargement of interlobular space,inflammatory infiltration,fat necrosis,hemorrhage,and the destruction or deletion of pancreatic acini were observed in the SAP groups;while in the SAP+PL groups,the enlargement of interlobular space was larger than in the SAP groups,but there was no significantly changes(P>0.05) for the inflammatory infiltration,fat necrosis,hemorrhage,and the destruction or deletion of pancreatic acini between them;there was only dispersive regional edema of interlobular space in the SO group.The total histopathologic scores were significantly increased in SAP groups and PL groups compared to SO group(P<0.05),and there was no statistically significant differences between SAP groups and PL groups(P>0.05).Conclusion This study suggests that,in the early stage of SAP,interrupted peritoneal lavage combined with antibiotics reduces significantly the blood levels of LPS、TNF-α、IL-6 and IL-8(P<0.05),and it maybe contributed to the reduction of the systemic damage to promote SAP get better and improve the prognosis of SAP.