摘要
目的通过研究重症急性胰腺炎大鼠应用甘草甜素后其肿瘤坏死因子-α、白介素-1β、白介素- 6、核转录因子-κB、肠粘膜ICAM-1的动态变化,探讨甘草甜素对急性重症胰腺炎治疗的可能作用机制。
方法150只健康雄性Sprague-Dawley大鼠,体重160-200g,随机(随机表法)分为假手术组(S组)、重症急性胰腺炎(P组)和重症急性胰腺炎甘草甜素治疗组(GL组),每组50只。每组10只,检测72h内的生存时间,P组和GL组采用戊巴比妥钠腹腔注射麻醉下逆行胰胆管穿刺泵推5%牛磺胆酸钠建立SAP模型。GL组大鼠在术后腹腔内注射甘草甜素(100mg/kg)。分别于术后3h、6h、12h、24h四个时间点采用腹主动脉放血法将大鼠分批处死(每组10只),全自动生化分析仪检测血淀粉酶,采用放射免疫法测血清炎症细胞因子TNF-α、IL-1β、IL-6的水平;取胰腺组织免疫组化检测胰腺组织NF-κBp65活性,同时进行胰腺组织病理学评分的测定,取回肠组织免疫组化检测回肠黏膜ICAM-1的表达,同时进行回肠黏膜组织病理学评分的测定,并观察甘草甜素预处理后上述各指标的变化。应用SPSS16.0版统计软件进行分析,主要统计指标进行正态性检验和方差齐性检验,各统计数据均以( X±s)表示;多组均数间的比较采用one-wayANOVA检验,多组均数间的两两比较采用DunnettT3法, P<0.05为差异显著有统计学意义。
结果(1)大鼠生存时间GL组与P组相比,GL组的生存时间明显延长,P=0.021(P<0.05)差异具有统计学意义。血清血淀粉、TNF-α、IL-1β、IL-6含量变化:与S组相比,P组、GL组各时间点均明显升高(P<0.005),P组与GL组相比,GL组低于同时间点P组,但TNF-α、IL-1β在3h差异无统计学意义,分别为P=0.280、P=0.350 (P>0.05),血淀粉、IL-6在3h差异有统计学意义,分别为P=0.023、P=0.024 (P<0.05)但在6h、12h、18h均有显著性差异.其中6h分别:P=0.019、P=0.012、P=0.022、P=0.018 (P<0.05).12h分别为:P=0.027、P=0.024、P=0.015、P=0.013 (P<0.05),24h分别为:P=0.027、P=0.025、P=0.032、P=0.015 (P<0.05)。
(2)胰腺组织、肠粘膜病理学评分:与S组相比,P组、GL组各时间点均明显升高,差异有统计学意义(P<0.05),与P组相比,GL组低于同时间点P组,在3h差异无统计学意义,分别为P=0.261、P=0.378(P>0.05)但在6h、12h、18h有显著性差异,胰腺组织病理比较,分别为6h:P=0.025、12h:P=0.021、24h:P=0.019(P<0.05)肠粘膜组织病理比较,分别为6h:P=0.032、12h:P=0.022、24h:P=0.017(P<0.05)。
(3)胰腺组织NF-κBp65,肠粘膜组织ICAM-1活性的变化:与S组相比,P组、GL组各时间点均明显升高,差异有统计学意义(P<0.05).胰腺组织NF-κBp65:P组与GL组相比,在24h差异无统计学意义P=0.280(P>0.05),但在3h、6h、12h差异有显著性,分别为:3h:P=0.016、6h:P=0.022、12h:P=0.025(P<0.05)。肠粘膜组织ICAM-1:P组与GL组相比,在3h差异无统计学意义,P=0.248(P>0.05)但在6h、12h、24h差异有显著性,分别为:6h:P=0.025、12h:P=0.018、24h:P=0.022(P<0.05)。
结论1.甘草甜素可降低SAP大鼠血清中TNF-α、IL-1β、IL-6浓度,抑制胰腺组织中NF-κBp65、肠粘膜组织ICAM-1的活性,具有抑制炎症作用。
2.甘草甜素具有糖皮质激素样作用,可以减轻重症胰腺炎炎症反应,这可能为今后治疗SAP提供了一条新途径。
Objectives To research the changes of the tumor necrosis factor-α, IL-1β, interleukin-6, nuclear factor-κB, ICAM-1 of the mucosa after the application of glycyrrhizin in rats with severe acute pancreatitis, To study the possible effect mechanism of glycyrrhizin on severe acute pancreatitis
Methods 150 healthy male Sprague-Dawley rats weighing 160-200g, which were randomly divided into sham operationgroup( S group) , severe acute pancreatitis group(P group) and the treatment group with glycyrrhizin of the severe acute pancreatitis (GL group),each group are 50. , the ten detect the survival time of the 72h with each group, P group and GL group are established the SAP model by Puncturing retrograde cholangiopancreatography and pumping 5% sodium taurocholate with intraperitoneal injecting anesthesia through abdominal of the Sodium pento-barbital.Rats in GL group were injected glycyrrhizin after surgery(100mg/kg) ,and at 3h, 6h, 12h, 24h four time points rats were sacrificed in groups using abdominal aortic phlebotomy (n = 10), serum amylas measured by automatic biochemical analyzer ,serum inflammatory cytokines IL-1β, IL-6, TNF-αlevels measured by radioimmunoassay, the activity of NF-κBp65 in pancreatic were detected by immunohistochemistry , at the same time, the scores of pancreatic tissue pathology were detected,the expression of ICAM-1 in ileal mucosa adhesion were detected by immunohistochemistry , at the same time the scores of ileal mucosa pathology were detected and observed the changes of the above indicators after glycyrrhizin pretreatment .All Statistical analysis were performed with SPSS16.0.Main Statistical indicators should be taken normality and homogeneity test,each data were expressed with Mean±standard deviation value,and Several more groups were compared by one-way ANOVA,two groups were analysed by DunnettT3 ,.If the P is less than 0.05 ,It is considered statistical significance. Results
Results (1) the survival time of GL groupRats compared with the P group, the survival time of GL group were significantly prolonged , P = 0.021 (P<0.05), difference was statistically significant. The content change of the blood serum amylase, TNF-α、IL-1βand IL-6 is: Compared with S group , the P group and GL group were significantly increased at each time point, P group compared with the GL, GL group was lower than SAP group at the same time point,but TNF-α、IL-1βshowed no significant difference in 3 h point, they respectively are :P = 0.280、P = 0.350 (P>0.05), serum amylase、IL-6 showed significantly difference in 3h point , they respectively are :P = 0.023、P = 0.024(P<0.05),but they are significantly different in 6h, 12h, 18h point . In the 6h point they respectively are: P = 0.019、P = 0.012、P = 0.022、P = 0.018 (P<0.05) .In the 6h point they respectively are: P = 0.027、P = 0.024、P = 0.015.、P = 0.013 (P<0.05) In the 6h point they respectively are : P = 0.027.、P = 0.025、P = 0.032、P = 0.015 (P<0.05)
(2) the pathological scores of pancreas and intestinal mucosa: P group and GL group were significantly higher than S group at each time point, they are significantly different (P<0.05), Compared with the P group, GL group was lower than P group at the same time point ,It showed no significant difference in 3 h point, they respectively are : P = 0.261、P = 0.378 (P>0.05) but they are significantly different in 6h, 12h, 18h point, the comparison of pancreatic pathology :they respectively are:6h: P = 0.025、12h: P = 0.021、24h: P = 0.019 (P<0.05) .the comparison of intestinal pathology :they respectively are 6h : P = 0.032、12h: P = 0.022、24h: P = 0.017 (P<0.05).
(3) the activity change of NF-κB p65 in pancreatic and ICAM-1 in intestinal mucosa: P group and GL group were significantly higher than S group at each time point, they are significantly different (P<0.05), the activity change of NF-κB p65 in pancreatic,GL group was lower than P group at the same time point ,It showed no significant difference in 24h point P = 0.280 (P>0.05), but they are significantly different in 3h、6h、12h point, they respectively are:3h: P = 0.016、6h: P = 0.022、12h: P = 0.025 (P<0.05) .the activity change of ICAM-1 in intestinal mucosa ,GL group was lower than P group at the same time point ,It showed no significant difference in 3h point P = 0.248 (P>0.05), but they are significantly different in 6h、12h、24h point, they respectively are:6h: P = 0.025、12h: P = 0.018、24h: P = 0.022 (P<0.05) .
Conclusions 1. Glycyrrhizin may reduce the concentration of TNF-α, IL-1β, IL-6 in the serum of the Severe acute pancreatitis and have inhibit inflammation.
2.Glycyrrhizin with glucocorticoid-like effects, may reduce inflammation in severe acute pancreatitis, which may treat SAP to provide a new way in the future
引文
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