摘要
目的:探究谷氨酰胺(Gln)强化肠内营养对重症急性胰腺炎(SAP)大鼠调节性T细胞/辅助性T细胞17(Treg/Th17)平衡的影响及机制。方法:80只大鼠随机分为假手术组(Sham)、SAP组、肠内营养组(EN)、Gln联合肠内营养组(Gln+EN),每组20只。采用逆行胰胆管注射5%牛黄胆酸钠构建SAP大鼠模型;造模4 h后,各组采用鼻空肠置管法进行营养治疗,EN组、Gln+EN组分别给予肠内营养液、添加Gln的肠内营养液;Sham组、SAP组给予等体积0.9%氯化钠溶液。48 h后,HE染色检测胰腺组织病理损伤;ELISA检测血清淀粉酶、脂肪酶、IL-6、TNF-α、IL-10、IL-17水平;流式细胞术检测脾脏和胰腺Th17、Treg细胞比例;RT-qPCR检测脾脏Foxp3、CTLA-4、GITR、RORγt、IL-17A mRNA水平;免疫组化染色、免疫荧光染色、Western blot检测胰腺髓过氧化物酶(MPO)、CD68、IL-6、TNF-α、IL-10、IL-17水平。结果:Sham组大鼠胰腺结构正常,无出血及坏死;SAP组胰腺结构破坏,有明显出血、坏死及炎症浸润;EN组和Gln+EN组胰腺仅存在少量出血、坏死及炎症浸润。与Sham组相比,SAP组胰腺组织学评分、血清淀粉酶、脂肪酶水平升高,胰腺MPO+中性粒细胞和CD68+巨噬细胞增多,血清和胰腺IL-6、TNF-α、IL-17水平升高,IL-10水平降低,Th17细胞百分比、RORγt、IL-17A mRNA水平升高,Treg细胞百分比、Treg/Th17、Foxp3、CTLA-4、GITR mRNA水平降低(均P<0.05);与SAP组相比,EN组和Gln+EN组胰腺组织学评分、血清淀粉酶、脂肪酶水平降低,胰腺MPO+中性粒细胞和CD68+巨噬细胞减少,血清和胰腺IL-6、TNF-α、IL-17水平降低,IL-10水平升高,Th17细胞百分比、RORγt、IL-17A mRNA水平降低,Treg细胞百分比、Treg/Th17、Foxp3、CTLA-4、GITR mRNA水平升高(均P<0.05);且Gln+EN对SAP的治疗作用优于EN(均P<0.05)。结论:Gln强化肠内营养能够调控SAP大鼠Treg/Th17平衡,降低机体全身炎症反应。
Objective:To explore the effect and mechanism of glutamine(Gln)enhanced enteral nutrition on regulatory T cells/T helper cell 17(Treg/Th17)balance in rats with severe acute pancreatitis(SAP).Methods:Eighty rats were randomly divided into Sham group,SAP group,enteral nutrition group(EN)and Gln+EN group,with 20 rats in each group.SAP model were constructed by retrograde injection of 5%sodium taurocholate into the pancreatic duct.After 4 hours of modeling,nasojejunostomy catheterization was used to nutritional treatment,EN group and Gln+EN group were given enteral nutrition solution and added Gln enteral nutrition solution,respectively;sham group and SAP group were given equal volume of 0.9%NaCl solution.After 48 hours,HE staining was used to detect pathological damages in pancreatic tissue;ELISA was used to detect amylase,lipase,IL-6,TNF-α,IL-10 and IL-17 levels in serum;flow cytometry was used to detect Th17 and Treg cells proportion in spleen and pancreas;RT-qPCR was used to detect Foxp3,CTLA-4,GITR,RORγt,IL-17A mRNA levels in spleen;immunohistochemical staining,immunofluorescence staining,Western blot were used to detect MPO,CD68,IL-6,TNF-α,IL-10,IL-17 levels in pancreas.Results:Pancreatic structure in Sham group was normal,without bleeding or necrosis;pancreatic structure in SAP group was disrupted with significant bleeding,necrosis and inflammatory infiltration;pancreatic structure in EN group and Gln+EN group were only a small amount of bleeding,necrosis and inflammatory infiltration.Compared with Sham group,histological score,amylase and lipase levels in serum in SAP group were increased,and MPO+neutrophils and CD68+macrophages in pancreas were increased,IL-6,TNF-α,IL-17 levels in serum and pancreas were increased,IL-10 level was decreased,Th17 cell percentage,RORγt,IL-17A mRNA levels were increased,while Treg percentage,Treg/Th17,Foxp3,CTLA-4 and GITR mRNA levels were decreased(all P<0.05);compared with SAP group,histological score,amylase and lipase levels in serum in EN group and Gln+EN group were decreased,and MPO+neutrophils and CD68+macrophages in pancreas were decreased,IL-6,TNF-α,IL-17 levels in serum and pancreas were decreased,IL-10 level was increased,Th17 cell percentage,RORγt,IL-17A mRNA levels were decreased,while Treg cell percentage,Treg/Th17,Foxp3,CTLA-4 and GITR mRNA levels were increased(all P<0.05);and Gln+EN has a better therapeutic effect on SAP than EN(all P<0.05).Conclusion:Gln enhanced enteral nutrition can regulate Treg/Th17 balance in SAP rats and reduce systemic inflammatory response.
作者
朱晓丽
刘宇
张翼
汪雯
ZHU Xiaoli;LIU Yu;ZHANG Yi;WANG Wen(Department of Nephrology,Beijing Anzhen Nanchong Hospital,Capital Medical University&Nanchong Central Hospital,Nanchong 637000,China;Department of Neurology,Beijing Anzhen Nanchong Hospital,Capital Medical University&Nanchong Central Hospital,Nanchong 637000,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1951-1957,共7页
Chinese Journal of Immunology
