摘要
目的:探讨血必净注射液(XBJ)通过调控甲酰肽受体(FPRs)/核苷酸结合寡聚化结构域样受体3(NLRP3)炎症途径对重症急性胰腺炎急性肺损伤(SAP-ALI)的治疗作用.方法:将60只大鼠随机分为假手术组,SAP-ALI模型组,XBJ低、中、高剂量组(4、8、12 mL·kg-1),阳性药(BOC2,0.2 mg·kg-1)组.假手术组仅在开腹后轻轻翻动胰腺,即关闭腹腔,其余5组以5％牛磺胆酸钠(Na-Tc)胆胰管逆行注射制备SAP-ALI大鼠模型.XBJ及BOC2在造模前3 d和造模后0.5 h腹腔注射给药.造模6 h后腹主动脉采血,酶联免疫吸附测定法(ELISA)测定血浆中白细胞介素(IL)-1β、IL-6及肿瘤坏死因子(TNF)-α的表达情况;测量腹水量及胰腺、肺组织干湿重比;取胰腺、肺组织行苏木素-伊红(HE)染色观察病理变化并评分;采用免疫组化法检测肺脏组织甲酰肽受体(FPR)1、FPR2和NLRP3的蛋白表达水平;蛋白免疫印迹法(Western blot)检测肺脏组织中FPR1、FPR2及NLRP3的表达.实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织FPR1、FPR2及NLRP3 mRNA的表达.结果:与假手术组比较,SAP-ALI模型大鼠肺组织干湿重比值显著降低(P＜0.01),肺组织病变严重,病理评分显著上升(P＜0.01),肺脏组织FPR1、FPR2和NLRP3蛋白及mRNA表达量显著增多(P＜0.01).BOC2干预后,上述检测指标均显著逆转(P＜0.01).经XBJ治疗后,各剂量组取得了与BOC2 一致的效果.结论:XBJ可有效改善SAP-ALI大鼠肺脏炎症反应,减轻损伤,其机制可能与抑制肺脏组织FPRs和NLRP3的表达,降低IL-1β,同时拮抗炎症因子IL-6和TNF-α的释放有关.
Abstract
Objective:To explore the therapeutic effect of Xuebijing injection(XBJ)on severe acute pancreatitis induced acute lung injury(SAP-ALI)by regulating formyl peptide receptors(FPRs)/nucleotide-binding oligomerization domain-like receptor 3(NLRP3)inflammatory pathway.Methods:Sixty rats were randomly divided into a sham group,a SAP-ALI model group,low-,medium-,and high-dose XBJ groups(4,8,and 12 mL·kg-1),and a positive drug(BOC2,0.2 mg·1kg-1)group.For the sham group,the pancreas of rats was only gently flipped after laparotomy,and then the abdomen was closed,while for the remaining five groups,SAP-ALI rat models were established by retrograde injection of 5％sodium taurocholate(Na-Tc)via the biliopancreatic duct.XBJ and BOC2 were administered via intraperitoneal injection once daily for 3 d prior to modeling and 0.5 h after modeling.Blood was collected from the abdominal aorta 6 h after the completion of modeling,and the expression of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)in plasma was measured by enzyme-linked immunosorbent assay(ELISA).The amount of ascites was measured,and the dry-wet weight ratios of pancreatic and lung tissue were determined.Pancreatic and lung tissue was taken for hematoxylin-eosin(HE)staining to observe pathological changes and then scored.The protein expression levels of FPR1,FPR2,and NLRP3 in lung tissue were detected by the immunohistochemical method.Western blot was used to detect the expression of FPR1,FPR2,and NLRP3 in lung tissue.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of FPR1,FPR2,and NLRP3 in lung tissue.Results:Compared with the sham group,the SAP-ALI model group showed significantly decreased dry-wet weight ratio of lung tissue(P＜0.01),serious pathological changes of lung tissue,a significantly increased pathological score(P＜0.01),and significantly increased protein and mRNA expression levels of FPR1,FPR2,and NLRP3 in lung tissue(P＜0.01).After BOC2 intervention,the above detection indicators were significantly reversed(P＜0.01).After treatment with XBJ,the groups of different XBJ doses achieved results consistent with BOC2 intervention.Conclusion:XBJ can effectively improve the inflammatory response of the lungs in SAP-ALI rats and reduce damage.The mechanism may be related to inhibiting the expression of FPRs and NLRP3 in lung tissue,which thereby reduces IL-1β and simultaneously antagonize the release of inflammatory factors IL-6 and TNF-α.
国家科技期刊平台
NSTL
维普
万方数据