辣蓼对大肠杆菌性腹泻小鼠炎症因子和细胞色素P450酶表达的影响

doi:10.5246/jcps.2022.08.052

中国药学(英文版) ›› 2022, Vol. 31 ›› Issue (8): 622-633.DOI: 10.5246/jcps.2022.08.052

辣蓼对大肠杆菌性腹泻小鼠炎症因子和细胞色素P450酶表达的影响

黄嘉欢^#, 岳玲^#, 张铭儒, 杨全, 程轩轩^*()

广东药科大学中药学院, 国家中医药管理局岭南药材生产与开发重点研究室,国家中药材产业技术体系广州综合试验站, 广东省南药规范化种植与综合开发工程技术研究中心, 广东广州 510006

收稿日期:2022-01-14 修回日期:2022-02-16 接受日期:2022-03-29 出版日期:2022-09-03 发布日期:2022-09-03
通讯作者: 程轩轩
作者简介:
+ Tel.: +86-20-39352176, E-mail: gdyxycxx@126.com
基金资助:
Key Project at Central Government Level: The ability establishment of sustainable use for valuable Chinese medicine resources (Grant No. 2060302-2004-03).

Effects of Polygonum hydropiper on the expressions of inflammatory cytokines and cytochrome P450 enzymes in mice with E. coli-induced diarrhea

Jiahuan Huang^#, Ling Yue^#, Mingru Zhang, Quan Yang, Xuanxuan Cheng^*()

Key Laboratory of State Administration of Traditional Chinese Medicine for Production & Development of Cantonese Medicinal Materials, Guangzhou Comprehensive Experimental Station of National Industrial Technology System for Chinese Materia Medica, Guangdong Engineering Research Center of Good Agricultural Practice & Comprehensive Development for Cantonese Medicinal Materials, School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou 510006, China

Received:2022-01-14 Revised:2022-02-16 Accepted:2022-03-29 Online:2022-09-03 Published:2022-09-03
Contact: Xuanxuan Cheng
About author:
# These authors contributed equally to this article.

摘要/Abstract

摘要：

本文旨在探讨辣蓼水提液对大肠杆菌性腹泻小鼠炎症因子和细胞色素P450 (CYPs) 酶表达的影响。将BALB/c小鼠随机分为正常组、模型组、恩诺沙星组、辣蓼水提液5 g/kg组和10 g/kg组。腹腔注射致病性大肠杆菌建立小鼠腹泻模型, 于造模前两天各组给予相应药物灌胃, 模型组和正常组给予等量的生理盐水, 连续给药11天。HE染色法观察十二指肠和肝脏的病理组织学特征, ELISA法检测十二指肠和肝脏中炎症因子和CYPs主要亚型的含量, qRT-PCR和Western Blot法检测CYPs的mRNA及蛋白表达。结果表明, 辣蓼水提液可明显缓解致病性大肠杆菌诱导的小鼠肠道及肝脏损伤, 降低炎症因子的含量及mRNA表达, 对CYPs主要亚型的mRNA和蛋白表达水平有明显的调节作用。辣蓼对大肠杆菌性腹泻有明显的治疗作用, 其机制可能与抑制炎症因子释放、调节药物代谢酶CYPs的表达有关。

关键词: 辣蓼, 致病性大肠杆菌, 腹泻, 炎症因子, 细胞色素P450

Abstract:

In the present study, we aimed to investigate the effects of Polygonum hydropiper aqueous extract (PHE) on the expression levels of inflammatory cytokines and cytochrome P450 (CYPs) in mice with E. coli-induced diarrhea. BALB/c mice were randomly divided into the control group, model group, enrofloxacin-treated group, and two PHE-treated groups with different doses. The diarrhea model was established by intraperitoneal injection of enteropathogenic E. coli (EPEC) in mice. The enrofloxacin-treated group was given enrofloxacin at 5 mg/kg by intragastric gavage (i.g.) for 11 d. PHE-treated groups were given PHE at 5 g/kg and 10 g/kg by i.g. for 11 d. The histopathological characteristics of the duodenum and liver were observed by HE staining. The levels of inflammatory cytokines and CYPs in the duodenum and liver of mice were determined by ELISA. The mRNA and protein expressions of CYPs were determined by qRT-PCR and Western blotting analysis, respectively. The results showed that PHE could significantly alleviate the injury of the duodenum and liver induced by EPEC infection, reduce the contents and mRNA expressions of inflammatory cytokines, and regulate the mRNA and protein expressions of the major subtypes of CYPs. These findings indicated that PHE had an apparent therapeutic effect on EPEC-induced diarrhea, and its mechanism might be related to inhibition of inflammatory cytokines and regulation of CYPs.

Key words: Polygonum hydropiper L., Enteropathogenic E. coli, Diarrhea, Inflammatory cytokines, Cytochrome P450

Supporting:

黄嘉欢, 岳玲, 张铭儒, 杨全, 程轩轩. 辣蓼对大肠杆菌性腹泻小鼠炎症因子和细胞色素P450酶表达的影响[J]. 中国药学(英文版), 2022, 31(8): 622-633.

Jiahuan Huang, Ling Yue, Mingru Zhang, Quan Yang, Xuanxuan Cheng. Effects of Polygonum hydropiper on the expressions of inflammatory cytokines and cytochrome P450 enzymes in mice with E. coli-induced diarrhea[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(8): 622-633.

图/表 8

Table 1. Primer sequences.

Figure 1. Effects of PHE on histopathological characteristics of the duodenum (A) and liver (B) in experimental mice. (200 ×) Representative H&E-stained sections showed necrosis, nuclear pyknosis or fragmentation (red arrow), inflammatory cell infiltration and exudation, eosinophilic?cytoplasm (yellow arrow), capillary congestion, and tissue edema (black arrow), loose cytoplasm (green arrow).

Table 2. Contents of TNF-α, IL-1β, IL-6, and LIF in duodenum and liver of mice.

Table 3. Contents of CYPs in duodenum and liver of experimental mice.

Table 4. Relative mRNA expression levels of TNF-α, IL-1β, IL-6, and LIF in duodenum and liver of mice.

Table 5. Relative mRNA expressions of CYPs in the duodenum of mice.

Table 6. Relative mRNA expressions of CYPs in the liver of mice.

Figure 2. Effects of PHE on the protein expressions of CYP1A1 and CYP1B1. The expressions of CYP1A1 and CYP1B1 at the protein level in the duodenum (A) and liver (C) of experimental mice were detected using Western blotting analysis. Results showed the expressions of CYP1A1 and CYP1B1 in the duodenum (B) and liver (D) of experimental mice. Data are presented as mean ± SD (n = 5). *P < 0.05 and **P < 0.01 vs. CON group; #P < 0.05 and ##P < 0.01 vs. MOD.

参考文献 37

[1]	Luo, Q.Z. Study on prevention and treatment of calf diarrhea with traditional Chinese medicine. Chin. J. Trad. Vet. Sci. 2019, 3, 55.
[2]	Wang, Z.B. Reasons and control measures of diarrhea in broilers. Mod. Anim. Husb. 2017, 9, 27.
[3]	Wei, H.; Li, J.J.; Wang, W.H. Reasons for colibacilosis in pig and control measures. Shaanxi J. Agr. Sci. 2018, 64, 92–95.
[4]	Chen, C.R.; Han, M.M.; Zhang, N.J.; Wang, W.B.; Yao, Y.B.; Xiao, S.; Li, Y. Association of biofilm formation with drug-resistance and virulence of Escherichia coli isolated from diarrhea piglet. Microbiol. China. 2016, 43, 2234–2241.
[5]	Seki, E.; Schnabl, B. Role of innate immunity and the microbiota in liver fibrosis: crosstalk between the liver and gut. J. Physiol. 2012, 590, 447–458.
[6]	Hakansson, A.; Molin, G. Gut microbiota and inflammation. Nutrients. 2011, 3, 637–682.
[7]	Chinese, VPC Veterinary pharmacopoeia of the people’s republic of China. Volume III. China Agricultural Press. 2011, 235.
[8]	Huang, H.H.; Zhen, H.S. Research progress of Polygonum hydropiper L. in recent years. Chin. J. Ethnomed. Ethnopharm. 2013, 22, 38–40.
[9]	Yu, X.; Li, W. Research progress on the regulation of cytochrome P450 expression in inflammatory diseases. J. Dalian Med. Univ. 2017, 39, 87–91.
[10]	Song, F.X.; Wang, R.; Yuan, Y.F. Application progress in cytochrome P450 enzymes and research methods of drug metabolism. Med. Recapit. 2017, 23, 665–669.
[11]	Tian, M.Y.; Fan, J.H.; Zhuang, Z.W.; Dai, F.; Wang, C.Y.; Hou, H.T.; Ma, Y.Z. Effects of silymarin on p65 NF-κB, p38 MAPK and CYP450 in LPS-induced hoof dermal inflammatory cells of dairy cows. BMC Vet. Res. 2019, 15, 127.
[12]	Chen, Y.; Li, S.; Su, P.; Zhang, H.J.; Zhang, G.P.; Ye, Z.G. Anti-diarrhea effect and mechanism of traditional Chinese medicine. Chin. J. Exp. Tradit. Med. Formulae. 2019, 25, 203–211.
[13]	Hu, N.; Huang, Y.J.; Gao, X.J.; Li, S.; Yan, Z.X.; Wei, B.; Yan, R. Effects of dextran sulfate sodium induced experimental colitis on cytochrome P450 activities in rat liver, kidney and intestine. Chem. Biol. Interact. 2017, 271, 48–58.
[14]	Xiao, Z.H.; Luo, X.Y.; Cheng, X.X.; Zheng, S.J.; Yang, Q.; Yang, H.W. Effect of polygoni hydropiperis herba on intestinal mucosal repair in diarrhea mice induced by escherichia coli. Chin. J. Exp. Tradit. Med. Formulae. 2018, 24, 120–126.
[15]	Kawauchi, S.; Nakamura, T.; Yasui, H.; Nishikawa, C.; Miki, I.; Inoue, J.; Horibe, S.; Hamaguchi, T.; Tanahashi, T.; Mizuno, S. Intestinal and hepatic expression of cytochrome P450s and mdr1a in rats with indomethacin-induced small intestinal ulcers. Int. J. Med. Sci. 2014, 11, 1208–1217.
[16]	Zheng, S.F.; Jin, X.; Chen, M.H.; Shi, Q.X.; Zhang, H.F.; Xu, S.W. Hydrogen sulfide exposure induces jejunum injury via CYP450s/ROS pathway in broilers. Chemosphere. 2019, 214, 25–34.
[17]	Xie, F.; Ding, X.X.; Zhang, Q.Y. An update on the role of intestinal cytochrome P450 enzymes in drug disposition. Acta Pharm. Sin. B. 2016, 6, 374–383.
[18]	Almazroo, O.A.; Miah, M.K.; Venkataramanan, R. Drug metabolism in the liver. Clin. Liver Dis. 2017, 21, 1–20.
[19]	Gideon, D.A.; Kumari, R.; Lynn, A.M.; Manoj, K.M. What is the functional role of N-terminal transmembrane helices in the metabolism mediated by liver microsomal cytochrome P450 and its reductase? Cell Biochem. Biophys. 2012, 63, 35–45.
[20]	Zhou, X.Y.; Tian, LX; Huang, Q.; Liang, H.P. Research progress on the relationship between cytochrome P4501A1 (CYP1A1) and immune diseases. Infection Inflammation Repair. 2017, 18, 120–124.
[21]	Gonzalez, J.; Marchand-Geneste, N.; Giraudel, J.L.; Shimada, T. Docking and QSAR comparative studies of polycyclic aromatic hydrocarbons and other procarcinogen interactions with cytochromes P450 1A1 and 1B1. SAR QSAR Environ. Res. 2012, 23, 87–109.
[22]	Liu, M.C.; Meng, Q.Q.; Cui, J.H.; Li, S.S. Recent research progress on the CYP1B1 inhibitors. Prog. Mod. Biomed. 2017, 17, 4190–4196.
[23]	Udomsuk, L.; Juengwatanatrakul, T.; Putalun, W.; Jarukamjorn, K. Bimodal action of miroestrol and deoxymiroestrol, phytoestrogens from Pueraria candollei var. mirifica, on hepatic CYP2B9 and CYP1A2 expressions and antilipid peroxidation in mice. Nutr. Res. 2012, 32, 45–51.
[24]	Pan, Y.J.; Omori, K.; Ali, I.; Tachikawa, M.; Terasaki, T.; Brouwer, K.L.R.; Nicolazzo, J.A. Altered expression of small intestinal drug transporters and hepatic metabolic enzymes in a mouse model of familial Alzheimer’s disease. Mol. Pharm. 2018, 15, 4073–4083.
[25]	Zhang, L.Q.; Ba, L.; Yu, S.; Ha, S. Advance in cytochrome P450 2J subfamily. Prog. Vet. Med. 2014, 35, 84–89.
[26]	Madanayake, T.W.; Fidler, T.P.; Fresquez, T.M.; Bajaj, N.; Rowland, A.M. Cytochrome P450 2S1 depletion enhances cell proliferation and migration in bronchial epithelial cells, in part, through modulation of prostaglandin E(2) synthesis. Drug Metab. Dispos. Biol. Fate Chem. 2012, 40, 2119–2125.
[27]	Zhao, C. Expression detection and phenotypic analysis of cytochrome CYP3A11 knock down mice. D. Southwest Univ. 2011.
[28]	Thesseling, F.A.; Hutter, M.C.; Wiek, C.; Kowalski, J.P.; Rettie, A.E.; Girhard, M. Novel insights into oxidation of fatty acids and fatty alcohols by cytochrome P450 monooxygenase CYP4B1. Arch. Biochem. Biophys. 2020, 679, 108216.
[29]	Barć, J.; Karpeta, A.; Gregoraszczuk, E.Ł. Action of halowax 1051 on enzymes of phase I (CYP1A1) and phase II (SULT1A and COMT) metabolism in the pig ovary. Int. J. Endocrinol. 2013, 2013, 590261.
[30]	Lian-Sheng, W. Progress in down-regulation of cytochrome P450 enzyme activities and mechanisms during inflammation. Chin. J. Pharm. Toxicol. 2010, 24, 236–240.
[31]	Gandhi, A.; Moorthy, B.; Ghose, R. Drug disposition in pathophysiological conditions. Curr. Drug Metab. 2012, 13, 1327–1344.
[32]	Monshouwer, M.; Witkamp, R.F. Cytochromes and cytokines: changes in drug disposition in animals during an acute phase response: a mini-review. Vet. Q. 2000, 22, 17–20.
[33]	Lee, C.M.; Pohl, J.; Morgan, E.T. Dual mechanisms of CYP3A protein regulation by proinflammatory cytokine stimulation in primary hepatocyte cultures. Drug Metab. Dispos. Biol. Fate Chem. 2009, 37, 865–872.
[34]	Stejskalova, L.; Pavek, P. The function of cytochrome P450 1A1 enzyme (CYP1A1) and aryl hydrocarbon receptor (AhR) in the placenta. Curr. Pharm. Biotechnol. 2011, 12, 715–730.
[35]	Li, F.; Zhu, W.F.; Gonzalez, F.J. Potential role of CYP1B1 in the development and treatment of metabolic diseases. Pharmacol. Ther. 2017, 178, 18–30.
[36]	Zhao, Y.Y.; Yao, X.P.; Jiao, T.Y.; Tian, J.N.; Gao, Y.; Fan, S.C.; Chen, P.P.; Jiang, Y.M.; Zhou, Y.Y.; Chen, Y.X.; Yang, X.; Huang, M.; Bi, H.C. Schisandrol B promotes liver enlargement via activation of PXR and YAP pathways in mice. Phytomedicine. 2021, 84, 153520.
[37]	Ye, Z.; Liu, Z.P.; Henderson, A.; Lee, K.; Hostetter, J.; Wannemuehler, M.; Hendrich, S. Increased CYP4B1 mRNA is associated with the inhibition of dextran sulfate sodium-induced colitis by caffeic acid in mice. Exp. Biol. Med. (Maywood ). 2009, 234, 605–616.

辣蓼对大肠杆菌性腹泻小鼠炎症因子和细胞色素P450酶表达的影响

Effects of Polygonum hydropiper on the expressions of inflammatory cytokines and cytochrome P450 enzymes in mice with E. coli-induced diarrhea

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 37

相关文章 12

编辑推荐

Metrics

本文评价

[1]	卢磊, 邱成英, 陈谦. 莲花清瘟颗粒联合头孢噻肟钠对小儿肺炎患儿疗效及血清炎症因子的影响[J]. 中国药学(英文版), 2023, 32(11): 947-953.
[2]	李睿, 孔燕茹. 利格列汀对初诊2型糖尿病患者炎症因子和动脉硬化的影响[J]. 中国药学(英文版), 2021, 30(8): 692-698.
[3]	张烨, 郑丹萍, 水梦洋, 刘晔, 刘晓岩, 王银叶. 新化合物W026B通过抑制炎性细胞因子的产生减轻缺血性脑损伤, 与tPA合用作用增强[J]. 中国药学(英文版), 2018, 27(10): 675-685.
[4]	杨会霞, 陈恳, 梁舒瑶, 翟所迪, 易湛苗. 合用CPY450酶抑制剂对伏立康唑安全性和药代动力学的影响的系统评价[J]. 中国药学(英文版), 2017, 26(3): 202-211.
[5]	杨倩, 陈刚, 杨洋, 蔡雪婷, 庞中化, 胡春萍, 张双全, 曹鹏. 刺芒柄花素通过诱导Nrf2表达缓解DSS诱导的小鼠溃疡性结肠炎[J]. 中国药学(英文版), 2016, 25(3): 178-188.
[6]	侯凤琴, 孙新婷, 王贵强. 成人粪白细胞1-14/HPF的非血便急性感染性腹泻患者病原及抗生素应用的探讨[J]. 中国药学(英文版), 2015, 24(2): 121-127.
[7]	林珠灿, 张龙, 张睿卓, 黄安玉, 郭素华. 基于炎症细胞模型的景天三七醋酸乙酯部位抗炎活性及其HPLC指纹图谱的研究[J]. 中国药学(英文版), 2015, 24(10): 647-653.
[8]	侯凤琴, 王艳, 李俊, 刘颖, 王贵强. 北京成人急性腹泻的诊治与指南存差距[J]. 中国药学(英文版), 2014, 23(2): 132-137.
[9]	侯凤琴, 孙新婷, 王贵强^*. 教学医院不合理应用抗菌药物治疗成人急性感染性腹泻调查[J]. , 2010, 19(3): 229-234.
[10]	孔德涛, 凌笑梅^, 韩方斌, 龚京莉, 葛泽梅, 李润涛, 崔景荣^*. TM208在大鼠肝微粒体中代谢产物与细胞色素P450亚型代谢酶鉴别研究[J]. , 2008, 17(1): 30-34.
[11]	周江泉^*, 汤致强. 大鼠肝微粒体代谢研究槲皮素对CYP1A2,CYP2E1,CYP3A2活性的影响及抑制机制[J]. , 2005, 14(4): 231-236.
[12]	周江泉, 汤致强^*. 肿瘤酶学研究中细胞色素P450活化前体药物的基因治疗[J]. , 2005, 14(1): 1-9.