Effect of the warming and heat-clearing method on the percentage of regulatory T lymphocytes/T helper 17 cells in rats with acute-on-chronic liver failure
-
摘要:
目的观察清温并用法对慢加急性肝衰竭(ACLF)大鼠调节性T淋巴细胞(Treg)/辅助性T淋巴细胞(Th) 17表达的影响,探索其抗肝衰竭的作用机制。方法 SD大鼠140只随机分为正常组、模型组、清法组、温法组及清温并用法组。采用牛血清白蛋白致敏建立免疫性肝纤维化大鼠模型,然后以D-氨基半乳糖+脂多糖攻击建立ACLF模型。观察24 h内各组大鼠血清肝功能(ALT、TBil)、肝组织病理学改变及外周血Treg、Th17细胞数,并计算Treg/Th17比值。计量资料多组间比较釆用One-Way ANOVA比较,进一步两两比较选用LSD-t法检验。结果与正常组比较,模型组大鼠ALT、TBil在各时间点明显升高(P值均<0. 01),肝小叶结构被破坏,肝细胞见桥接及片状坏死,汇管区可见炎细胞浸润,外周血Treg、Th17细胞比例明显升高(P <0. 01),且Th17高于Treg,Treg/Th17比值明显降低(P <0. 01)。与模型组比较,清法组、温法组、清温并用法组ALT、TBil明显降低(P <0. 05),肝小叶结构较完整,肝细胞可见小片状、点状坏死,门区少量...
-
关键词:
- 肝功能衰竭 /
- T淋巴细胞,调节性 /
- T淋巴细胞,辅助诱导 /
- 温法 /
- 清法
Abstract:Objective To investigate the effect of the warming and heat-clearing method on the percentage of regulatory T (Treg) /T helper 17 (Th17) cells in rats with acute-on-chronic liver failure (ACLF) and the mechanism of its anti-liver failure activity. Methods A total of 140 Sprague-Dawley rats were randomly divided into normal group, model group, heat-clearing group, warming group, and warming + heat-clearing group. A rat model of immune liver fibrosis was established by sensitization with bovine serum albumin, and then an ACLF model was established by D-galactosamine + lipopolysaccharide attack. Serum liver function parameters [alanine aminotransferase (ALT) and total bilirubin (TBil) ], liver pathological changes, and number of peripheral Treg and Th17 cells were observed within 24 hours, and Treg/Th17 ratio was calculated. One-way ANOVA was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in ALT and TBil at each time point (all P < 0. 01) , with the damage of lobular structure, bridging and piecemeal necrosis in hepatocytes, and inflammatory cell infiltration in the portal area. Compared with the normal group, the model group also had significant increases in the percentage of peripheral Treg and Th17 cells (P < 0. 01) , and the percentage of Th17 cells was higher than that of Treg cells; there was a significant reduction in Treg/Th17 ratio in the model group (P < 0. 01) . Compared with the model group, the heat-clearing group, the warming group, and the warming + heat-clearing group had significant reductions in ALT and TBil (P< 0. 05) , with a more complete lobular structure, patchy and spotted necrosis in hepatocytes, and mild fibrous tissue hyperplasia at the portal area; these three groups also had significant reductions in the percentage of peripheral Treg and Th17 cells (P < 0. 05) , and Th17 cells had a greater reduction than Treg cells; the warming + heat-clearing group had significantly greater reductions than the heat-clearing group and the warming group (P < 0. 05) . Conclusion〗 The warming and heat-clearing method can regulate Treg/Th17 imbalance in rats with ACLF and is one of the mechanisms of its anti-liver failure activity.
-
[1] Liver Failure and Artificial Liver Group, Chinese Society of Infectious Diseases, Chinese Medical Association; Severe Liver Diseases and Artificial Liver Group, Chinese Society of Hepatology, Chinese Medical Association. Guidelines for diagnosis and treatment liver failure[J]. Chin J Clin Infect Dis, 2012, 5 (6) :321-327. (in Chinese) 中华医学会感染病学分会肝衰竭与人工肝学组, 中华医学会肝病学分会重型肝病与人工肝学组.肝衰竭诊治指南 (2012年版) [J].中华临床感染病杂志, 2012, 5 (6) :321-327. [2] RICHARD M, VICENTE A. Acute-on-chronic liver failure:A new clinical entity[J]. Clin Gastroenterol Hepatol, 2015, 13 (5) :836-841. [3] DENG H. Correlation between Treg/Th17 imbalance and the liver injury caused by systemic inflammatory response and oxidative stress in children with sepsis[J]. J Hanan Med Coll, 2017, 23 (21) :2968-2971. (in Chinese) 邓辉.脓毒症患儿Treg/Th17失衡与全身炎症反应、氧化应激所致肝损伤的相关性[J].海南医学院学报, 2017, 23 (21) :2968-2971. [4] CHEN B, WANG J, PENG J, et al. Effect of Wen-Yang Jie-Du Hua-Yu granules on TLR4 and NF-κB expression of rat hepatic failure model with intestinal endotoxemia[J]. World Sci Technol Modern Tradit Chin Med, 2013, 15 (5) :818-824. (in Chinese) 陈斌, 王杰, 彭杰, 等.温阳解毒化瘀颗粒对肝衰竭肠源性内毒素血症大鼠TLR4、NF-κB表达的影响[J].世界科学技术-中医药现代化, 2013, 15 (5) :818-824. [5] ZHU WF, SUN KW, CHEN B, et al. Effect of Wenyang Jiedu Huayu prescription on intestinal bacteria in patients with HBV related liver failure[J]. Chin J Integr Tradit West Med Liver Dis, 2014, 24 (4) :214-216. (in Chinese) 朱文芳, 孙克伟, 陈斌, 等.温阳解毒化瘀方对HBV相关肝衰竭患者肠道菌群的影响[J].中西医结合肝病杂志, 2014, 24 (4) :214-216. [6] ZHU QG, FANG BW, ZHU JN, et al. Animal models of bovine serum albumin-induced immune liver fibrosis[J]. Chin Jf Integr Tradition Western Med Liver Dis, 1992, 2 (1) :20-24. (in Chinese) 朱起贵, 方步武, 竺稽能, 等.牛血清白蛋白免疫性肝纤维化动物模型的研究[J].中西结合肝病杂志, 1992, 2 (1) :20-24. [7] LIU XH, CHEN Y, ZHANG J, et al. Human serum albumin and a rat's model for d-galactosamine/lipopolysacchar ide induced acute-on-chronic liver failure[J]. Sci Technol Rev, 2007, 25 (18) :32-37. (in Chinese) 刘旭华, 陈煜, 张晶, 等.大鼠慢加急性肝衰竭实验模型的建立及其病理机制初步研究[J].科技导报, 2007, 25 (18) :32-37. [8] XU Y, LIN H, ZHENG W, et al. Matrine ameliorates adriamycin-induced nephropathy in rats by enhancing renal function and modulating Th17/Treg balance[J]. Eur J Pharmacol, 2016, 15 (7) :491-501. [9] YANG C, CUI F, CHEN LM, et al. Correlation between Th17 and nTreg cell frequencies and the stages of progression in chronic hepatitis B[J]. Mol Med Rep, 2016, 13 (1) :853-862. [10] MING M, LUO Z, LV S, et al. Inactivated Mycobacterium phlei inhalation ameliorates allergic asthma through modulating the balance of CD4+CD25+regulatory T and Th17 cells in mice[J]. Iran J Basic Med Sci, 2016, 19 (9) :953-959. [11] SHI WJ, JIA JT, LI CD, et al. Expression and significance of Th17/Treg cytokines in patients with hepatitis B-related acute-on-chronic liver failure[J]. Chin Hepatol, 2016, 21 (2) :92-94. (in Chinese) 施文娟, 贾瑾堂, 李彩东, 等. Th17/Treg细胞因子在乙型肝炎慢加急性肝衰竭患者中的表达及意义[J].肝脏, 2016, 21 (2) :92-94. [12] GAO HB, WANG XM, LIU BR, et al. Features of regulatory T cells and T helper 17 cells and their association with pre-C/BCP mutations in HBV in patients with HBV-related acute-on-chronic liver failure[J]. Chin Hepatol, 2018, 23 (1) :18-21. (in Chinese) 高海兵, 王香梅, 刘宝荣, 等. HBV相关慢加急性肝衰竭患者Treg细胞、Th17细胞表达特点及其与HBV前C区/BCP区变异的相关性[J].肝脏, 2018, 23 (1) :18-21. [13] ZHU L, XI Y, ZHAO L, et al. Intervenient study of Shengjiang San on Th17/Treg imbalances and related regulatory factors in sepsis patients[J]. J Clin Emerg, 2017, 18 (6) :406-409. (in Chinese) 朱亮, 奚耀, 赵雷, 等.升降散对脓毒症患者Th17/Treg失衡及相关调节因子的干预作用[J].临床急诊杂志, 2017, 18 (6) :406-409. [14] JIA L, WU J, QIU ZJ, et al. Effects of Yupingfeng Powder on balance of Th17/Treg in allergic rhinitis mice[J]. China J Tradit Chin Med Pharma, 2016, 31 (8) :3260-3262. (in Chinese) 贾翎, 吴珺, 邱泽计, 等.玉屏风散对过敏性鼻炎小鼠Th17/Treg平衡的影响[J].中华中医药杂志, 2016, 31 (8) :3260-3262. [15] WU LM, ZHAO JF. Role of Treg and Th17 cells in the pathogenesis of hepatitis B-induced acute-on-chronic liver failure[J].J Pract Hepatol, 2017, 20 (5) :639-642. (in Chinese) 吴丽蒙, 赵久法. Treg和Th17细胞在慢加急性乙型肝炎肝衰竭发病机制中的作用研究进展[J].实用肝脏病杂志, 2017, 20 (5) :639-642. [16] KAN YT, GAN JH, SUN W, et al. Changes in regulatory T cells and T helper 17 cells in patients with HBV-related acute-onchronic liver failure and related clinical significance[J]. Chin Hepatol, 2016, 21 (2) :89-91. (in Chinese) 阚延婷, 甘建和, 孙蔚, 等. HBV相关慢加急性肝衰竭患者Treg、Th17变化及其与临床相关性[J].肝脏, 2016, 21 (2) :89-91. [17] YU SJ, JIANG R, MAZZU YZ, et al. Epigallocatechin-3-gallate prevents triptolide-induced hepatic injury by restoring the Th17/Treg balance in mice[J]. Am J Chin Med, 2016, 44 (6) :1221-1236. [18] XUE S, CHEN Z, YIN Z, et al. Changes in circulating Foxp3+regulatory T cells and interleukin-17-producing T helper cells during HBV-related acute-on-chronic liver failure[J]. World J Gastroenterol, 2014, 20 (26) :8558-8571. [19] MICROAL B. Systemic inflammation, and multiorgan dysfunction in decompensated cirrhosis:Evidence for a nonfunctional mechanism of hepatorenal syndrome[J]. Hepatology, 2017, 11 (3) :242-244. 期刊类型引用(10)
1. 李晏杰,王娜,王沙,唐鲲,毛德文,石清兰,龙富立. HBV相关性肝衰竭发病中的Th细胞免疫调控机制及中医药干预的研究进展. 内蒙古中医药. 2023(11): 134-138 . 百度学术
2. 霍瑞静,张翠丽,李静,何晨熙,李帅,高健. NF-κB信号通路对慢性乙型肝炎患者Th17/Treg相关细胞因子水平变化的影响. 中华医院感染学杂志. 2022(17): 2570-2573 . 百度学术
3. 黄杏,刘文志,陈斌. 清温并用法治疗肝衰竭的中医理论及作用机制探讨. 中国医学创新. 2021(10): 164-167 . 百度学术
4. 陈斌,丁秀丽,彭杰,黎运芳,王若宇,周意. 清温并用法对慢加急性肝衰竭肠源性内毒素血症大鼠结肠组织FoxP3, ROR-γt表达的影响. 中国实验方剂学杂志. 2020(02): 19-25 . 百度学术
5. 谭年花,陈斌,彭杰,朱文芳,张涛,周意,王若宇. 乙肝相关性慢加急性肝衰竭“阳黄-阴阳黄-阴黄”不同证候分型患者Treg/Th17的表达. 中国实验方剂学杂志. 2020(02): 33-38 . 百度学术
6. 陈斌,黎运芳,张涛,朱文芳,彭杰,张茜茜. 清温并用法对慢加急性肝衰竭大鼠肝组织Treg/Th17型细胞因子及转录因子的影响. 中华中医药杂志. 2020(09): 4618-4622 . 百度学术
7. 李薇,于佳卉,王新婷,王春梅,李贺,陈建光,孙靖辉. 北五味子总木脂素对环磷酰胺诱导的免疫功能低下小鼠的调节作用. 吉林大学学报(医学版). 2019(02): 294-299+472 . 百度学术
8. 陈斌,彭杰,张涛,朱文芳,张茜茜,丁秀丽,黎运芳. 清温并用法对慢加急性肝衰竭大鼠结肠组织调节性T淋巴细胞/辅助性T淋巴细胞17细胞因子的影响. 湖南中医药大学学报. 2019(05): 573-577 . 百度学术
9. 张龙跃,张序瑞. 赤芍解毒汤保留灌肠治疗慢加急性肝衰竭临床研究. 系统医学. 2019(13): 138-140 . 百度学术
10. 梁琼,赵隽,苏静,梁琳,张吉发. 凉血化瘀解毒法对急性肝衰竭模型大鼠免疫功能、Toll样受体及VEGF表达的影响. 中国中医急症. 2019(10): 1700-1704 . 百度学术
其他类型引用(5)
-