基于Toll样受体4/核因子-κB信号通路研究莪术醇抗肝纤维化的分子机制

郑洋; 王嘉孺; 刘露露; 王佳慧; 赵铁建

doi:10.3969/j.issn.1001-5256.2020.07.013

基于Toll样受体4/核因子-κB信号通路研究莪术醇抗肝纤维化的分子机制

DOI: 10.3969/j.issn.1001-5256.2020.07.013

1. 广西中医药大学赛恩斯新医药学院医学系
2. 广东医科大学护理学院
3. 广西中医药大学第一附属医院教学部
4. 广西中医药大学基础医学院生理教研室

基金项目:

国家自然科学基金项目(81960751,81660705)；广西中青年教师科研基础能力提升项目(2020KY59009)；广西壮瑶药重点实验室(GXZYZZ2019-1)；广西中医药大学一流学科(2019XK141)；

详细信息

中图分类号: R285.5
计量
- 文章访问数: 1465
- HTML全文浏览量: 40
- PDF下载量: 158
- 被引次数: 0
出版历程
- 出版日期: 2020-07-20

Molecular mechanism of the anti-liver fibrosis effect of curcumol: An analysis based on the TLR4/NF-κB signaling pathway

1. Department of Medicine, Faculty of Chinese Medicine Science, Guangxi University of Chinese Medicine, Nanning 530021, China
2. School of Nursing, Guangdong Medical University, Dongguan, Guangdong 523000, China
3. Department of Education, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530022, China
4. Department of Physiology, School of Basic Medical Sciences, Guangxi University of Chinese Medicine, Nanning 530021, China

Research funding:

摘要

摘要:
目的研究莪术醇对肝窦内皮细胞TLR4/NF-κB信号通路的作用,以此来分析莪术醇抗肝纤维化的分子机制。方法将50只清洁级KM小鼠随机分为空白组（n=10）、模型组（n=19）以及莪术醇组（n=21）,除去建模过程中死亡的小鼠,最后每组各10只;细胞分为空白对照组、阳性对照组、莪术醇干预组以及PDTC（NF-κB特异性抑制剂）组。用HE和Masson染色观察各组小鼠肝脏结构的改变,用Western Blot和实时荧光定量PCR（RT-PCR）检测各组细胞TLR4/NF-κB信号通路上关键分子TLR4和NF-κB mRNA和蛋白的表达水平,用免疫荧光检测各组细胞NF-κB的表达及入核情况。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 RT-PCR检测发现莪术醇干预组TLR4 mRNA和NF-κB mRNA表达水平较阳性对照组下降,差异均有统计学意义（P值均<0. 05）; Western Blot检测发现莪术醇干预组TLR4和磷酸化NF-κB的表达水平较阳性对照组下降,差异均有统计学意义（P值均<0. 05）;免疫荧光检测发现莪术醇干预组NF-κ...
- 肝硬化 /
- 莪术醇 /
- 肝窦内皮细胞 /
- Toll样受体4 /
- NF-κB /
- 信号传导
Abstract:
Objective To investigate the molecular mechanism of the anti-liver fibrosis effect of curcumol by observing the effect of curcumol on the TLR4/NF-κB signaling pathway in liver sinusoidal endothelial cells. Methods A total of 50 mice were randomly divided into blank group,model group,and curcumol group,and cells were divided into blank control group,LPS positive control group,curcumol intervention group,and PDTC group. HE staining and Masson staining were used to observe the change in liver structure; Western blot and quantitative real-time PCR( RT-PCR) were used to measure the protein and mRNA expression of the key molecules TLR4 and NF-κB in the TLR4/NF-κB signaling pathway; immunofluorescence assay was used to observe the expression and nuclear import of NF-κB in cells. A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups. Results RT-PCR showed that compared with the positive control group,the curcumol intervention group had significant reductions in the mRNA expression of TLR4 and NF-κB( both P < 0. 05). Western blot showed that compared with the positive control group,the curcumol intervention group had significant reductions in the expression of TLR4 and NF-κB( both P <0. 05). Immunofluorescence assay showed that compared with the positive control group,the curcumol intervention group had significant improvement in NF-κB nuclear import. Conclusion Curcumol can exert an anti-liver fibrosis effect possibly by inhibiting the activity of the TLR4/NF-κB signaling pathway.
- liver cirrhosis /
- curcumol /
- liver sinusoidal endothelial cells /
- Toll-like receptor 4 /
- NF-kappa B /
- signal transduction

HTML全文

参考文献(30)

[1]ZHAO J,QI YF,YU YR.The role of oxidative stress in the development of liver fibrosis[J].J Clin Hepatol,2019,35(9):2067-2071.(in Chinese)赵杰，齐永芬，鱼艳荣．氧化应激在肝纤维化发生发展中的作用[J]．临床肝胆病杂志，2019,35(9):2067-2071．

[2]ALTAMIRANO-BARRERA A,BARRANCO-FRAGOSO B,MENDEZ-SANCHEZ N.Management strategies for liver fibrosis[J].Ann Hepatol,2017,16(1):48-56．

[3]ZOUBEK ME,TRAUTWEIN C,STRNAD P.Reversal of liver fibrosis:From fiction to reality[J].Best Pract Res Clin Gastroenterol,2017,31(2):129-141．

[4]SØRENSEN KK,SIMON-SANTAMARIA J,MCCUSKEY RS,et al.Liver sinusoidal endothelial cells[J].Compr Physiol,2015,5(4):1751-1774．

[5]YANG L,JIANG YY.Research progress of hepatic sinusoidal endothelial cells and hepatic sinusoidal capillarization[J].Infect Dis Info,2010,23(3):183-186.(in Chinese)杨丽，江宇泳．肝窦内皮细胞与肝窦毛细血管化研究进展[J]．传染病信息，2010,23(3):183-186．

[6]SVISTOUNOV D,WARREN A,MCNERNEY GP,et al.The relationship between fenestrations,sieve plates and rafts in liver sinusoidal endothelial cells[J].PLo S One,2012,7(9):e46134．

[7]DU YQ,WU J,YANG DL.Role of liver sinusoidal endothelial cells in the development and progression of liver fibrosis and related therapeutic strategy[J].J Clin Hepatol,2017,33(3):438-444.(in Chinese)杜艳芹，吴珺，杨东亮．肝窦内皮细胞在肝纤维化发生发展中的机制及相关治疗策略[J]．临床肝胆病杂志，2017,33(3):438-444．

[8]TERATANI T,TOMITA K,SUZUKI T,et al.A high-cholesterol diet exacerbates liver fibrosis in mice via accumulation of free cholesterol in hepatic stellate cells[J].Gastroenterology,2012,142(1):152-164．

[9]ZHU XJ,ZHANG F,KONG DS,et al.Progress in lipopolysaccharide/Toll-like receptor 4 signaling transduction in hepatic fibrosis[J].Chin J Pharmacol Toxicol,2013,27(1):106-109.(in Chinese)朱晓静，张峰，孔德松，等．脂多糖/Toll样受体4信号转导与肝纤维化的研究进展[J]．中国药理学与毒理学杂志，2013,27(1):106-109．

[10]PAIK YH,LEE KS,LEE HJ,et al.Hepatic stellate cells primed with cytokines upregulate inflammation in response to peptidoglycan or lipoteichoic acid[J].Lab Invest,2006,86(7):676-686．

[11]TANG SC,LIAO PY,HUNG SJ,et al.Topical application of glycolic acid suppresses the UVB induced IL-6,IL-8,MCP-1 and COX-2inflammation by modulating NF-κB signaling pathway in keratinocytes and mice skin[J].J Dermatol Sci,2017,86(3):238-248．

[12]IE CI,APPA RS,HILDEN I,et al.Rat liver sinusoidal endothelial cells(LSECs)express functional low density lipoprotein receptor-related protein-1(LRP-1)[J].J Hepatol,2011,55(6):1346-1352．

[13]ISHAK K,BAPTISTA A,BIANCHI L,et al.Histological grading and staging of chronic hepatitis[J].J Hepatol,1995,22(6):696-699．

[14]SEBASTIANI G,GKOUVATSOS K,PANTOPOULOS K.Chronic hepatitis C and liver fibrosis[J].World J Gastroenterol,2014,20(32):11033-11053．

[15]TACKE F,TRAUTWEIN C.Mechanisms of liver fibrosis resolution[J].J Hepatol,2015,63(4):1038-1039．

[16]ZHENG Y,WANG JH,LIANG TJ,et al.Effect of curcumol on Rho-ROCK signaling pathway in hepatic stellate cells[J].Chin Hosp Pharm J,2019,39(15):1517-1520.(in Chinese)郑洋，王佳慧，梁天坚，等．莪术醇对肝星状细胞Rho-ROCK信号通路作用的实验研究[J]．中国医院药学杂志，2019,39(15):1517-1520．

[17]BRAET F,WISSE E.Gentle palpating liver sinusoidal endothelial cells reveals the dynamic behavior and formation of fenestrae:A new window for biomedical research[J].Hepatology,2018,67(6):2460-2461．

[18]ELVEVOLD K,SMEDSRD B,MARTINEZ I.The liver sinusoidal endothelial cell:A cell type of controversial and confusing identity[J].Am J Physiol Gastrointest Liver Physiol,2008,294(2):g391-g400．

[19]SCHAFFNER F,POPER H.Capillarization of hepatic sinusoids in man[J].Gastroenterology,1963,44:239-242．

[20]HILSCHER MB,SEHRAWAT T,ARAB JP,et al.Mechanical stretch increases expression of CXCL1 in liver sinusoidal endothelial cells to recruit neutrophils,generate sinusoidal microthombi,and promote portal hypertension[J].Gastroenterology,2019,157(1):193-209．

[21]NI Y,LI JM,LIU MK,et al.Pathological process of liver sinusoidal endothelial cells in liver diseases[J].World J Gastroenterol,2017,23(43):7666-7677．

[22]DELEVE LD,MARETTI-MIRA AC.Liver sinusoidal endothelial cell:An update[J].Semin Liver Dis,2017,37(4):377-387．

[23]KONG LJ,LI H,DU YJ,et al.Vatalanib,a tyrosine kinase inhibitor,decreases hepatic fibrosis and sinusoidal capillarization in CCl4-induced fibrotic mice[J].Mol Med Rep,2017,15(5):2604-2610．

[24]KAUR S,ROBERTS DD.CD47 applies the brakes to angiogenesis via vascular endothelial growth factor receptor-2[J].Cell Cycle,2011,10(1):10-12．

[25]LIN YT,CHEN LK,JIAN DY,et al.Visfatin promotes monocyte adhesion by upregulating ICAM-1 and VCAM-1 expression in endothelial cells via activation of p38-PI3K-Akt signaling and subsequent ROS production and IKK/NF-κB activation[J].Cell Physiol Biochem,2019,52(6):1398-1411．

[26]SHI X,WANG S,LUAN H,et al.Clinopodium chinense attenuates palmitic acid-induced vascular endothelial inflammation and insulin resistance through TLR4-Mediated NF-κB and MAPK pathways[J].Am JChin Med,2019,47(1):97-117．

[27]MENON D,COLL R,O'NEILL LA,et al.GSTO1-1 modulates metabolism in macrophages activated through the LPS and TLR4 pathway[J].J Cell Sci,2015,128(10):1982-1990．

[28]ZHU Q,ZOU L,JAGAVELU K,et al.Intestinal decontamination inhibits TLR4 dependent fibronectin-mediated cross-talk between stellate cells and endothelial cells in liver fibrosis in mice[J].J Hepatol,2012,56(4):893-899．

[29]GUO J,LOKE J,ZHENG F,et al.Functional linkage of cirrhosispredictive single nucleotide polymorphisms of Toll-like receptor 4 to hepatic stellate cell responses[J].Hepatology,2009,49(3):960-968．

[30]SHI H,DONG L,DANG X,et al.Effect of chlorogenic acid on LPS-induced proinflammatory signaling in hepatic stellate cells[J].Inflamm Res,2013,62(6):581-587.

施引文献

资源附件(0)

访问统计