连翘对重症急性胰腺炎大鼠肝组织中NF-κB和Foxp3表达的影响

Effects of Forsythia suspensa on expression of NF-κB and Foxp3 during liver injury in rats with severe acute pancreatitis

目的探讨核因子NF-κB和Foxp3在重症急性胰腺炎(SAP)肝损伤中的作用及连翘对其表达活性的影响。方法雄性Wistar大鼠80只,随机分成假手术组(SO组)、SAP组和干预组,其中干预组分连翘高、中、低剂量组和阳性对照组(PDTC)。牛磺胆酸钠溶液在胰胆管远端注射造模,SO和SAP组于术后3、6、12 h,干预组于术后12 h处死大鼠,分别留取标本。测各组血淀粉酶(AMY)、ALT及TNFα水平,鲎试剂法测血浆内毒素水平,流式细胞术测外周血Treg百分数,对胰腺及肝脏进行病理学检查及评分,RT-PCR法检测肝脏组织中NF-κBmRNA和Foxp3mRNA表达量。组间比较采用单因素方差分析,进一步进行多重比较,采用LSD法进行统计学处理,各指标间相关性分析采用直线相关分析。结果与SO组比较,SAP组中各项指标均随时间升高,于12 h达高峰。与SAP12 h组相比,干预组(大鼠死亡率为0)肝脏组织中的NF-κBmRNA和Foxp3mRNA表达明显降低(P<0.01),与Treg呈正相关(r=0.738,P<0.01)。随连翘剂量增加,AMY、ALT及TNFα水平均明显降低,肝脏和胰腺组织炎症明显减轻,高剂量组和阳性对照组相比较无明显差异(P>0.05)。结论 NF-κB的激活参与SAP肝损伤的发生,连翘能显著降低NF-κB的活性及肝脏组织中NF-κBmRNA和Foxp3mRNA的表达,减轻SAP肝损伤的严重程度。

Objective To investigate the roles of nuclear factor - kappa B （ NF - KB） and forkhead box P＇3 （ Foxp3 ） during liver injury in rats with severe acute pancreatitis （SAP） and the effects of Forsythia suspensa on their expression. Methods Eighty nlale Wistar rats were randomly allocated into sham operation （SO） group, SAP group, and intervention group. The intervention group was further divided into high -dose, middle -dose, and low -dose Forsythia suspensa subgroups and positive control （PDTC） subgroup. A rat model was induced by injecting sodium tauro- cholate into the bile - pancreatic duet. The rats in SO and SAP groups were sacrificed at 3, 6, and 12 h after operation, and those in intervention group were sacrificed at 12 h after operation. The serum levels of amylase （Amy）, alanine aminotransferase （ALT）, and tumor necrosis factor （TNF） ot were measured. The eudotoxin content in plasma was determined using a limulus reagent. The percentage of Treg cells among peripheral blood lymphocytes was measured by flow cytometty. The pathological examination and scoring of the liver and pancreas were performed. The mRNA expression levels of NF - KB and Foxp3 in liver tissue were measured by RT - PCR. One - way analysis of variance was used for comparison among groups ; multiple comparison was performed by LSD test ; the indices were subjected to linear correlation analysis. Results Compared with those in the SO group, all the indices in the SAP group increased over time and reached the peak levels at 12 h after operation. Compared with the rats in SAP group that were sacrificed at 12 h after operation, the intervention group （mortality = 0） had significantly decreased InRNA expression of NF - KB and Foxp3 in liver tissue （P 〈0.01 ）. The NF - KB mRNA level was positively correlated with the percentage of Treg cells （r =0. 738, P 〈 0.01 ）. As the dose of Forsythia suspensa rose, Amy, ALT, and TNFa levels decreased significantly, and the inflammation of liver and pancreas was significantly alleviated. There were no significant differences between the high -dose Forsythia suspensa subgroup and PDTC subgroup （P 〉 0. 05 ）. Conclusion Activation of NF - KB contributes to the liver injury in rats with SAP. Forsythia suspensa can significantly reduce the activation of NF - KB and the mRNA expression of NF - KB and Foxp3 in liver tissue, thus alleviating the liver injury in rats with SAP.