大黄素对重症急性胰腺炎小鼠Treg/Th17平衡的影响及机制研究
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摘要
目的:探讨大黄素对重症急性胰腺炎小鼠Treg/Th17平衡的影响及可能的机制。方法:将60只C57BL/6小鼠按照随机数表法分为3组:正常对照组(n=20)、模型对照组(n=20)和药物组(n=20),采用雨蛙素联合脂多糖法制备重症急性胰腺炎小鼠模型。模型构建成功后,药物组腹腔注射2.5 mg/(kg·d)大黄素,模型对照组和正常对照组均给予同等剂量的生理盐水。给药后48 h,将小鼠麻醉后处死。ELISA法检测血清中淀粉酶、脂肪酶、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、二胺氧化酶(diamine oxidase,DAO)和白细胞介素-10(interleukin-10,IL-10)水平;流式细胞术法检测脾脏组织中调节性T细胞(regulatory T-cell,Treg)及辅助性Th17细胞(T helper cell 17,Th17)比例;q-PCR法检测脾脏组织叉头状/翼状螺旋转录因子3(forkhead or winged helix transcription,FOXP3)、视黄酸相关的孤儿受体γt(retinoid acid related orphan receptor gamma t,RORγt)和Toll样受体4(Toll-like receptor 4,TLR4)mRNA的表达。结果:与正常对照组比较,模型对照组血清脂肪酶、血清淀粉酶、TNF-α、DAO、IL-10水平及Foxp3、RORγt和TLR4 mRNA表达量明显升高;Treg及Th17细胞数均升高,但Th17细胞上升比例高,Treg/Th17降低。与模型对照组比较,药物组血清脂肪酶、血清淀粉酶、TNF-α、DAO、IL-10水平及Foxp3、RORγt和TLR4 m RNA表达量明显降低。Treg及Th17细胞数均有所降低,Treg/Th17升高,均具有统计学意义(P<0.05)。结论:大黄素可明显降低TNF-α、DAO和IL-10的表达,缓解炎症应激和氧化应激反应,同时可调节Treg/Th17平衡,改善免疫紊乱,从而发挥治疗重症急性胰腺炎的作用。
Objective:To investigate the effect of emodin on regulatory T cell(Treg)/T helper 17 cell(Th17) balance in mice with severe acute pancreatitis and the possible mechanism. Methods:A total of 60 C57 BL/6 mice were divided into normal control group,model control group,and drug group using a random number table,with 20 mice in each group. Cerulein combined with lipopolysaccharide was used to establish a mouse model of severe acute pancreatitis. After the model was established successfully,the mice in the drug group were given intraperitoneally injected with emodin at a dose of 2.5 mg/kg/day,and those in the model control group and the normal control group were given normal saline at the same dose. The mice were sacrificed after anesthesia at 48 hours after administration. ELISA was used to measure the serum levels of amylase,lipase,tumor necrosis factor-α(TNF-α),diamine oxidase(DAO),and interleukin-10(IL-10);flow cytometry was used to measure the percentages of Treg cells and Th17 cells in the spleen;q-PCR was used to measure the mRNA expression of forkhead or winged helix transcription factor(Foxp3),retinoid acid-related orphan receptor gamma t(RORγt),and Toll-like receptor 4(TLR4) in the spleen. Results:Compared with the normal control group,the model control group had significant increases in the serum levels of lipase,amylase,TNF-α,DAO,and IL-10,the m RNA expression of Foxp3,RORγt,and TLR4,and the number of Treg and Th17 cells,as well as a significantly greater increase in the percentage of Th17 cells and a significant reduction in Treg/Th17 ratio(all P<0.05). Compared with the model control group,the drug group had significant reductions in the serum levels of lipase,amylase,TNF-α,DAO,and IL-10,the mRNA expression of Foxp3,RORγt,and TLR4,and the number of Treg and Th17 cells and a significant increase in Treg/Th17 ratio(all P<0.05). Conclusion:Emodin can significantly reduce the expression of TNF-α,DAO,and IL-10 and relieve inflammatory stress and oxidative stress response. It can also regulate Treg/Th17 balance,improve immune disorders,and thus play a role in the treatment of severe acute pancreatitis.
Objective:To investigate the effect of emodin on regulatory T cell(Treg)/T helper 17 cell(Th17) balance in mice with severe acute pancreatitis and the possible mechanism. Methods:A total of 60 C57 BL/6 mice were divided into normal control group,model control group,and drug group using a random number table,with 20 mice in each group. Cerulein combined with lipopolysaccharide was used to establish a mouse model of severe acute pancreatitis. After the model was established successfully,the mice in the drug group were given intraperitoneally injected with emodin at a dose of 2.5 mg/kg/day,and those in the model control group and the normal control group were given normal saline at the same dose. The mice were sacrificed after anesthesia at 48 hours after administration. ELISA was used to measure the serum levels of amylase,lipase,tumor necrosis factor-α(TNF-α),diamine oxidase(DAO),and interleukin-10(IL-10);flow cytometry was used to measure the percentages of Treg cells and Th17 cells in the spleen;q-PCR was used to measure the mRNA expression of forkhead or winged helix transcription factor(Foxp3),retinoid acid-related orphan receptor gamma t(RORγt),and Toll-like receptor 4(TLR4) in the spleen. Results:Compared with the normal control group,the model control group had significant increases in the serum levels of lipase,amylase,TNF-α,DAO,and IL-10,the m RNA expression of Foxp3,RORγt,and TLR4,and the number of Treg and Th17 cells,as well as a significantly greater increase in the percentage of Th17 cells and a significant reduction in Treg/Th17 ratio(all P<0.05). Compared with the model control group,the drug group had significant reductions in the serum levels of lipase,amylase,TNF-α,DAO,and IL-10,the mRNA expression of Foxp3,RORγt,and TLR4,and the number of Treg and Th17 cells and a significant increase in Treg/Th17 ratio(all P<0.05). Conclusion:Emodin can significantly reduce the expression of TNF-α,DAO,and IL-10 and relieve inflammatory stress and oxidative stress response. It can also regulate Treg/Th17 balance,improve immune disorders,and thus play a role in the treatment of severe acute pancreatitis.
引文
[1]唐学军,王小云,彭晓斌.生长抑素联合肠内营养对老年急性胰腺炎患者肠通透性与免疫反应的影响[J].实用老年医学,2016,30(10):812-814.
[2]冼丽娜,杨远征,陈伟. VitB1联合新斯的明足三里注射治疗重症急性胰腺炎胃肠动力障碍的作用[J].海南医学,2017,28(24):4059-4060.
[3]齐文杰,张苗苗,文艳,等.重症急性胰腺炎大鼠舌组织小窝蛋白表达及大黄素干预的实验研究[J].北京中医药,2018,37(1):40-43.
[4]石占利,方堃,孙静,等.大黄素对重症急性胰腺炎大鼠胰腺细胞凋亡的干预作用[J].中华全科医学,2017,15(11):1830-1834.
[5] Lew D,Afghani E,Pandol S. Chronic pancreatitis:current status and challenges for prevention and treatment[J]. Dig Dis Sci,2017,62(7):1702-1712.
[6]郭静,张双双,牛国平.以3倍正常上限值为cut off值评价血清脂肪酶和淀粉酶对急性胰腺炎的诊断价值[J].检验医学与临床,2016,13(3):336-337,340.
[7]刘海珊,符照康,李永超,等.生大黄灌肠辅助治疗急性胰腺炎的临床疗效及对淀粉酶与炎性炎症因子的影响[J].辽宁中医杂志,2017,44(5):962-965.
[8]刘瑞霞,齐文杰.大黄素治疗重症急性胰腺炎的作用与机制研究进展[J].临床和实验医学杂志,2016,15(2):193-195.
[9]Rohan JD,Osman HG,Patel S. Severe acute pancreatitis attacks are associated with significant morbidity and mortality[J]. Curr Probl Surg,2014,51(9):370-372.
[10]杨宝晶,尤胜义,张志远,等.大黄素治疗重症急性胰腺炎肺损伤机制研究[J].中国全科医学,2016,19(24):2943-2947.
[11]陈静.急性胰腺炎患者外周血中Th17和Treg细胞及其细胞因子水平变化[J].空军医学杂志,2016,32(6):368-371.
[12]马镇,苏彧.早期腹腔灌洗对重症急性胰腺炎患者外周血中Th17/Treg细胞平衡的影响[J].中国急救医学,2016,36(9):829-833.
[13]张志远.大黄素治疗重症急性胰腺炎作用机制的研究[D].天津:天津医科大学,2015.
[14]杨宝晶. TLR4信号途经对重症急性胰腺炎Treg/Thl7平衡的影响及大黄素治疗作用机制的研究[D].天津:天津医科大学,2016.
[15]邱会芬,王雯,王忠永,等.白癜风患者外周血Th17和Treg细胞特异性转录因子的检测及意义[J].实用医学杂志,2016,32(17):2881-2883.
[16]杨小猛,赵丹,杜丽伟,等.习惯性流产患者绒毛和蜕膜中Toll样受体4与Treg/TH17细胞免疫平衡的相关性研究[J].临床检验杂志,2015,33(3):193-196.
[17]龚小军,郜朝霞,王传明.大黄素对急性胰腺炎大鼠相关炎性因子表达的影响[J].药物分析杂志,2015,35(12):2095-2098.
[2]冼丽娜,杨远征,陈伟. VitB1联合新斯的明足三里注射治疗重症急性胰腺炎胃肠动力障碍的作用[J].海南医学,2017,28(24):4059-4060.
[3]齐文杰,张苗苗,文艳,等.重症急性胰腺炎大鼠舌组织小窝蛋白表达及大黄素干预的实验研究[J].北京中医药,2018,37(1):40-43.
[4]石占利,方堃,孙静,等.大黄素对重症急性胰腺炎大鼠胰腺细胞凋亡的干预作用[J].中华全科医学,2017,15(11):1830-1834.
[5] Lew D,Afghani E,Pandol S. Chronic pancreatitis:current status and challenges for prevention and treatment[J]. Dig Dis Sci,2017,62(7):1702-1712.
[6]郭静,张双双,牛国平.以3倍正常上限值为cut off值评价血清脂肪酶和淀粉酶对急性胰腺炎的诊断价值[J].检验医学与临床,2016,13(3):336-337,340.
[7]刘海珊,符照康,李永超,等.生大黄灌肠辅助治疗急性胰腺炎的临床疗效及对淀粉酶与炎性炎症因子的影响[J].辽宁中医杂志,2017,44(5):962-965.
[8]刘瑞霞,齐文杰.大黄素治疗重症急性胰腺炎的作用与机制研究进展[J].临床和实验医学杂志,2016,15(2):193-195.
[9]Rohan JD,Osman HG,Patel S. Severe acute pancreatitis attacks are associated with significant morbidity and mortality[J]. Curr Probl Surg,2014,51(9):370-372.
[10]杨宝晶,尤胜义,张志远,等.大黄素治疗重症急性胰腺炎肺损伤机制研究[J].中国全科医学,2016,19(24):2943-2947.
[11]陈静.急性胰腺炎患者外周血中Th17和Treg细胞及其细胞因子水平变化[J].空军医学杂志,2016,32(6):368-371.
[12]马镇,苏彧.早期腹腔灌洗对重症急性胰腺炎患者外周血中Th17/Treg细胞平衡的影响[J].中国急救医学,2016,36(9):829-833.
[13]张志远.大黄素治疗重症急性胰腺炎作用机制的研究[D].天津:天津医科大学,2015.
[14]杨宝晶. TLR4信号途经对重症急性胰腺炎Treg/Thl7平衡的影响及大黄素治疗作用机制的研究[D].天津:天津医科大学,2016.
[15]邱会芬,王雯,王忠永,等.白癜风患者外周血Th17和Treg细胞特异性转录因子的检测及意义[J].实用医学杂志,2016,32(17):2881-2883.
[16]杨小猛,赵丹,杜丽伟,等.习惯性流产患者绒毛和蜕膜中Toll样受体4与Treg/TH17细胞免疫平衡的相关性研究[J].临床检验杂志,2015,33(3):193-196.
[17]龚小军,郜朝霞,王传明.大黄素对急性胰腺炎大鼠相关炎性因子表达的影响[J].药物分析杂志,2015,35(12):2095-2098.
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