重症急性胰腺炎大鼠免疫抑制点的研究  

The investigation on the turning point from over-inflammation to immunosuppression in rat severe acute pancreatitis model

作　　者：翟朝霞 刘超[1] 邓冬梅[1] 匡梅[1] 岑彦艳[1] 尚圣兰 覃容欣[1] 周红[1] ZHAI Zhaoxia LIU Chao DENG Dongmei KUANG Mei CEN Yanyan SHANG Shenglan QIN Rongxin ZHOU Hong(Department of Pharmacology, College of Pharmacy, Third Military Medical University, Chongqing 400038, Chin)

机构地区：[1]第三军医大学药学系药理学教研室,重庆400038

出　　处：《免疫学杂志》2017年第11期921-929,共9页Immunological Journal

基　　金：重庆市基础科学与前沿技术研究专项(cstc2015jcyjBX0049);国家自然科学基金(81673495);第三军医大学科技成果转化基金(2013XZH03)

摘　　要：目的研究重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠模型中是否存在从过度炎症反应状态向免疫抑制状态转变的病理生理过程以及免疫抑制转折点的发生时间。方法采用胰胆管逆行注射不同浓度牛磺胆酸钠的方法建立SAP大鼠模型;大鼠尾静脉注射不同浓度的铜绿假单胞菌(Pseudomonas aeruginosa,PA),观察大鼠7 d死亡率,筛选出PA亚致死剂量;然后在SAP建模后不同时间点给予亚致死剂量PA攻击,观察大鼠死亡率;在SAP建模后0 h、12 h、24 h给予PA攻击,ELISA检测各组大鼠血清、肝、肺及脾中TNF-α、IL-1β水平;在SAP建模后24 h给予PA攻击,五分类血液分析仪测定各组血常规,平板菌落计数的方法测定各组血清、肺及脾中细菌清除情况,ELISA检测各组大鼠血清、肝、肺及脾中TNF-α、IL-1β水平。结果采用胰胆管逆行注射5.0%牛磺胆酸钠的方法成功复制出48%死亡率的SAP大鼠模型;PA攻击大鼠的亚致死剂量为4.0×108CFU/kg;SAP大鼠在建模后不同时间点对PA的敏感性不同;SAP建模后24 h,大鼠对PA攻击呈现出高细菌敏感性,表现为血液中的高细菌负荷和低致炎细胞因子水平;此外,淋巴细胞和单核细胞数量和比例也呈现出显著降低状态。结论 SAP大鼠模型中存在从过度炎症反应状态向免疫抑制状态转变的病理生理过程;SAP大鼠模型建立后24 h是免疫抑制转折点。Severe acute pancreatitis（SAP） is a severe clinical condition with significant morbidity and mortality. Multiple organs dysfunction（MOD） and/or the systemic inflammatory response syndrome（SIRS） are the leading causes of SAP-related death. SAP shares a similar profile of inflammation with sepsis, but there is no systemic report about immunosuppression during SAP. Herein, whether there is immunosuppression to happen during SAP, and the turning point from over-inflammation to immunosuppression in SAP rat model was investigated in this study. SAP rat model was established by a standard retrograde infusion of sodium taurocholate solution into the biliopancreatic duct after laparotomy. Then the mortalities of rats were observed in SAP rats, which were challenged with Pseudomonas aeruginosa（PA） at different time-points after model establishment. In addition, the levels of TNF-α and IL-1β in each group were measured at 0, 12, and 24 h. And the levels of proinflammatory cytokines, bacterial loads, and immune cells were tested at 24 h. Data showed that SAP rats presented a different bacterial sensitivity to PA, and at 24 h after model establishment they had highest sensitivity than Sham rats. At this time-point, SAP rats challenged with PA presented higher bacterial loads and lower levels of pro-inflammatory cytokines. Additionally, the amounts and proportion of lymphocytes and monocytes significantly decreased,too. The above results suggested that there is immunosuppression in SAP rat model; and 24 h after the model establishment is the turning point from over-inflammation to immunosuppression.

关 键 词：重症急性胰腺炎 免疫抑制 脓毒症 铜绿假单胞菌 细胞因子 

分 类 号：R576[医药卫生—消化系统]