早期腹腔穿刺引流对重症急性胰腺炎大鼠腹腔巨噬细胞表型极化的影响  

Abdominal paracentesis drainage ameliorates severe acute pancreatitis through regulating peritoneal macrophage mediated inflammation in rats

作　　者：刘若鸿 刘春雨 张誉凡 孙红玉[2] 汤礼军 LIU Ruo-hong;LIU Chun-yu;ZHANG Yu-fan;SUN Hong-yu;TANG Li-jun(Graduate School of Army Medical University,Chongqing 400038,China;PLA Center of General Surgery &Pancreatic Injury and Repair Key Laboratory of Sichuan Province,Chengdu Military General Hospital,Chengdu 610083,China;physical Examination Center,Jiaotong Hospital Affiliated to Sichuan Provincial People's Hospital,Chengdu 611730,China)

机构地区：[1]陆军军医大学研究生院,重庆400038 [2]成都军区总医院全军普通外科中心,四川省胰腺损伤与修复重点实验室,成都610083 [3]四川省人民医院交通医院体检办,成都611730

出　　处：《解放军医学杂志》2018年第10期834-839,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81772001);国家临床重点专科建设项目(41792113)

摘　　要：目的探索早期腹腔穿刺引流(APD)对重症急性胰腺炎(SAP)大鼠腹腔巨噬细胞表型极化的影响。方法健康成年雄性SPF级SD大鼠36只,随机分为假手术组(Sham组)、SAP组和APD组,每组12只。SAP组和APD组大鼠采用5%牛磺胆酸钠胰胆管逆行注射法建立SAP模型,APD组大鼠右下腹留置腹腔引流管,Sham组大鼠不注射药物,仅在腹腔内留置引流管后关腹。术后12h取材,HE染色观察胰腺组织病理损伤程度并进行评分,ELISA法测定血清淀粉酶、脂肪酶、促炎因子和抗炎因子分泌情况,流式细胞术检测腹腔巨噬细胞表型极化情况,qPCR检测腹腔细胞中肿瘤坏死因子α(TNF-α)、诱导型一氧化氮合酶(iNOS)、CD163、CD206 mRNA的表达情况。结果与SAP组比较,APD组胰腺组织病理损伤明显减轻,病理评分明显降低(P<0.05),血清淀粉酶、脂肪酶、促炎因子TNF-α和白细胞介素1β(IL-1β)浓度明显降低,而抗炎因子IL-10、转化生长因子β(TGF-β)浓度明显升高(P<0.05)。与SAP组比较,APD组腹腔巨噬细胞中CD163阳性细胞明显增多,TNF-α阳性细胞明显减少(P<0.05),腹腔细胞中M2型巨噬细胞标记物CD163、CD206mRNA表达明显上调(P<0.05),而M1型巨噬细胞标记物TNF-α、iNOSmRNA的表达两组比较差异无统计学意义(P>0.05)。结论腹腔穿刺引流可通过增加腹腔巨噬细胞的M2极化抑制炎症反应,进而起到缓解SAP病情的作用。Objective To investigate the effect of early stage abdominal paracentesis drainage(APD) on phenotype transition of peritoneal macrophage in rats with severe acute pancreatitis(SAP). Methods A total of 36 adult male Sprague Dawley(SD) rats were randomized into SAP, APD, and sham-operated(Sham) groups(n=12). Pancreatitis in SAP and APD groups were induced by 5% sodium taurocholate retrograde injection. In the APD group, a drainage tube was placed in the lower right abdomen of rats with pancreatitis before closure. In the Sham group, rats didn’t receive the injection procedure, and a segment of the drainage tube was placed in the abdomen. The rats were sacrificed 12 hours after model established. The histopathological injury was evaluated by HE staining and scored using a previously described scoring system. The level of amylase and lipase, pro/anti-inflammatory mediators in serum, were determined by ELISA. The polarization of peritoneal macrophage was analyzed by flow cytometry. The mRNA expression of M1-associated protein tumor necrosis factor-α(TNF-α), inducible nitric oxide synthase(iNOS), CD163 and CD206 in peritoneal cells were detected by qPCR. Results The histopathological injury and its score of the pancreas in APD group decreased as well as the serum level of amylase, lipase, TNF-α, interleukin-1β(IL-1β) compared with SAP group, however, the level of anti-inflammatory cytokines interleukin-10(IL-10) and transforming growth factor-β(TGF-β) in plasma increased significantly(P<0.05). Moreover, compared with SAP group, in the APD group, the number of CD163(+) macrophages and the mRNA expression of M2-associated protein CD163 and CD206 increased in peritoneal cells, while the number of TNF-α(+) macrophages decreased(P<0.05). However, the mRNA expression of M1-associated protein TNF-α and iNOS did not show any significant difference between each group(P>0.05). Conclusion APD ameliorates SAP by enhancing the M2 polarization of peritoneal macrophage and inhibiting systematic inflammation.

关 键 词：重症急性胰腺炎 腹腔穿刺引流 巨噬细胞 极化 

分 类 号：R657.51[医药卫生—外科学]