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Efficacy of Sishen Wan(四神丸) on dinitrobenzene sulfonic acid-induced ulcerative colitis and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB signal pathway

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Journal of Traditional Chinese Medicine 2022年第4期42卷 565-575页

作者： ZHANG Zhaohua LIU Rong DU Nana ZHU Xiangdong School of Basic Medicine Gansu university of Chinese MedicineLanzhou 730000China College of Chinese Medicine Ningxia medical universityYinchuan 750000China Department of Thoracic Surgery Gansu Provincial Cancer HospitalLanzhou 730000China

OBJECTIVE:To investigate the therapeutic effect of Sishen Wan(四神丸,SSW)on ulcerative colitis(UC)induced by dinitrobenzene sulfonic acid and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB(TLR2/IRAK4/NF-κB)signaling pathway in colonic ***:In this study,120 Sprague–Dawley rats were randomly divided into blank and model *** experimental UC model in rats was established by subcutaneous injection of hydrocortisone+senna gavage for 21 d+dinitrobenzene sulfonic acid(DNBS)/ethanol solution *** successful model rats were randomly divided into the model group;mesalazine(0.36 g/kg)group;and high-,medium-,and low-dose SSW(24,12,and 6 g/kg)*** model and blank groups were gavaged with equal volumes of distilled water once a day for 21 *** general condition of the rats was observed,and the body mass,fecal properties,and occult blood were recorded for calculating the disease activity index(DAI)*** colonic tissue of the rats was collected,and its general morphology and pathological form were noted for obtaining the colonic mucosal injury index(CMDI)***-eosin staining was used to view the pathological changes of the colon tissue in each group,apoptosis of the cells was detected using terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling staining,and quantitative real-time polymerase chain reaction was used to measure the expressions of TLR2,myeloid differentiation primary response gene 88(My D88),IRAK4,and NF-κB p65 mRNA in the colon *** expressions of TLR2,My D88,IRAK4,and NF-κB p65 protein were detected using western blotting and immunohistochemistry assay,and the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the colon tissue were determined using enzyme linked immunosorbent ***:Compared with the blank group,the general condition of the model group was relatively *** DAI and CMDI scores of the model group increased signific

关键词： colitis,ulcerative toll-like receptor 2 interleukin-1 receptor-associated kinases NF-kappa B signal transduction therapeutic uses Sishen Wan

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Protection against hepatic ischemia/reperfusion injury via downregulation of toll-like receptor 2 expression by inhibition of Kupffer cell function

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World Journal of Gastroenterology 2005年第28期11卷 4423-4426页

作者： Jin-Xiang Zhang He-Shui Wu Yang Wang Hui Wang Jin-Hui Zhang Qi-Chang Zheng Department of Emergency Surgery Union Hospital Tongji Medical CollegeHuazhong University of Science and Technology Wuhan 430030Habei Province China Hereditary Department Tongji Medical CollegeHuazhong University of Science and Technology Wuhan 430030Habei Province China Department of General Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM： To elucidate the mechanism of liver protection by inhibition of Kupffer cells （KCs） function.METHODS： All the animals were randomly divided into three groups. Blockade group （gadolinium chloride solution （GdCl3） injection plus ischemia/reperfusion （I/R） injury）：GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group （saline solution injection plus I/R injury）： saline instead of GdCl3 as a control was injected as in the blockade group. Sham group： saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. toll-like receptor2 （TLR2） was immunostained with a goat antimouse polydonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-α （TNF-α） and alanine aminotransferase （ALT）, an indicator of liver function.I/R injury via downregulation of theexpression of TLR2.RESULTS： Compared to non-blockade group, CD^68＋ cells significantly reduced in blockade group （OPTDI, optical density integral）： 32.97±10.55 vs 185.65±21.88, P〈0.01） and the liver function impairment was relieved partially （level of ALT： 435.89±178.37 U/L vs 890.21±272.91 U/L,P〈0.01）. The expression of TLR2 protein in blockade group significantly decreased compared to that in non- group（method of immunohistochemistry, OPDTI： 75.74±17.44 vs 170.58±25.14, P〈0.01; method of Western blot, A value： 125.89±15.49 vs 433.91±35.53, P〈0.02）. The latter correlated with the variation of CD68 staining （r = 0.745,P〈0.05）. Also the level of portal vein TNF-α decreased in blockade group compared to that in non-blockade group （84.45±14.73 ng/L vs 112.32±17.56 ng/L, P〈0.05）, butwas still higher than that in sham group （84.45±14.73

关键词： toll-like receptor 2 Reperfusion injury Kupffer cell Liver

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Relationship between the expression of toll-like receptor 2 and 4 in mononuclear cells and postoperative acute lung injury in orthotopic liver transplantation

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Chinese Medical Journal 2009年第8期122卷 895-899页

作者： CHI Xin-jin CAI Jun LUO Chen-fang CHENG Nan HEI Zi-qing LI Shang-rong LUO Gang-jian Department of Anesthesiology Third Affiliated Hospital Sun Yat-sen University Guangzhou Guangdong 510630 China

Background The aim of this study was to investigate the potential relationship between the dynamic expression of toll-like receptor 2 and 4 （TLR2/4） in peripheral blood mononuclear cells as well as changes in serum concentration of inflammatory factors and acute lung injury （ALl） in patients after orthotopic liver transplantation （OLT）. Methods The peripheral blood samples of 27 patients （23 men and 4 women with ASA Ⅲ to Ⅳ） who received OLT were collected for measurement of TLR2/4 at T1 （after induction of anesthesia）, T2 （25 minutes after anhepatic phase）, T3 （3 hours after graft reperfusion） and T4 （24 hours after graft reperfusion）. The expression of TLR2/4 in mononuclear cells was measured by flow cytometry. The serum concentrations of tumor necrosis factor （TNF）-α, interleukin （IL）-1β and IL-8 were measured by enzyme-linked immunosorbent assay （ELISA）. Twenty-seven patients were assigned to ALl group （n=-9） and non-ALl group （n=-18） according to the diagnostic criteria of ALl. The expression of TLR2/4 in the ALl group or non-ALl group was analyzed. Results Compared to the non-ALl group, the volumes of blood loss, ascites, total output and transfused red blood cells were higher in the ALl group, and the anhepatic phase lasted longer （P 〈0.05, P 〈0.01）. The expression of TLR2/4 in mononuclear cells increased significantly at T3 and T4, and serum concentrations of TNF-α, IL-1β and IL-8 increased significantly too. There was no significant difference in Child-Turcotte-Pugh （CTP） scores between the ALl group and non-ALl group （P 〉0.05）. The expression of TLR2/4 in mononuclear cells increased significantly at T3 and T4 in the ALl group （P 〈0.05, P〈0.01）. A positive correlation was noted between the expression of TLR4 in mononuclear cells and the serum concentrations of TNF-α, IL-1β （P=0.041, P=0.046） in the ALl group. In the non-ALl group, statistical results showed that the expression level of TLR2/4 in mononuclear cells was not significantly different during the pe

关键词： orthotopic liver transplantation toll-like receptor 2 toll-like receptor 4 acute lung injury

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Ureaplasma urealyticum-derived lipid-associated membrane proteins introduce IL-6, IL-8, and TNF-α cytokines into human amniotic epithelial cells via toll-like receptor 2

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2018年第8期19卷 654-661页

作者： Guang-yong YE Ke-yi WANG Qiao-di GUI Min WANG Women's Hospital Zhejiang University School of Medicine Central Laboratory Hangzhou First People's Hospital Department of Clinical Laboratory Shaanxi Provincial People's Hospital

Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on toll-like receptors （TLRs）. Methods： LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells （HAECs） were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay （ELISA） and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results： LAMPs induced HAECs to produce inflammatory cytokines interleukin （IL）-6, IL-8, and tumor necrosis factor （TNF）-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor （anti-hTLR2-IgA）. Conclusions： LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.

关键词： Ureaplasma urealyticum Lipid-associated membrane protein Human amniotic epithelial cell toll-like receptor 2

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Different expression of dectin-1 and toll-like receptor 2 in the lungs of different immune status mice infected with Aspergillus fumigatus

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Chinese Medical Journal 2009年第17期122卷 2017-2021页

作者： YANG Jian-xun LIU Wan LU Qiao-yun WAN Zhe WANG Xiao-hong LI Ruo-yu Department of Dermatology Research Center of MedicalMycology Peking University First Hospital Beijing 100034China

Background Invasive pulmonary aspergillosis （IPA） is a severe and frequently fatal disease in patients receiving treatment with immunosuppressive agents such as cyclophosphamide. Aspergillus fumigatus （A. fumigatus） now is a leading cause of IPA. Dectin-1 and toll-like receptor 2 （TLR2） are important pattern recognition receptors involved in immune responses to A. fumigatus in vitro. However, the expression of the two receptors during the infection of A. fumigatus in vivo is not completely understood. The effects of cyclophosphamide treatment on the expression of the receptors need to be further studied. Methods We established different immune status in mice models with or without A. fumigatus infection. On days 1, 3 and 5 post inoculation, pulmonary tissues from mice of the different groups were harvested. Dectin-1 and TLR2 mRNA expression in the lungs of the mice were investigated by real-time PCR. The pulmonary fungal burden in the mice with A. fumigatus infection was also evaluated. Results In the immunocompetent mice, three days after A. fumigatus inoculation, dectin-1 and TLR2 expression increased markedly compared with the normal control group. Cyclophosphamide inhibited the clearance of pathogens and the expression of dectin-1 with or without A. fumigatus infection in the lungs as well. There was no statistical difference in TLR2 expression between the different immune status groups. Conclusions Our results suggest that in vivo, dectin-1 and TLR2 are activated during the experimental period which would provide a broad range of possibilities for a specific and effective inflammatory response to kill A. fumigatus. Inhibition of dectin-1 expression may be one of the mechanisms of cyclophosphamide in the development of IPA. Chin Med J 2009; 122（17）：2017-2021

关键词： Aspergillus fumigatus dectin-1 toll-like receptor 2 real-time polymerase chain reaction

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Pudilan Anti-inflammatory Oral Liquid and Organic Acid Component from Taraxaci Herba Attenuate Allergic Asthma in Young Mice Through toll-like receptor 2/toll-like receptor 4 Signaling Pathway

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World Journal of Traditional Chinese Medicine 2024年第3期10卷 294-302页

作者： Yu-Zhi Mao Chen-Zi Li Wei-Quan Bu Bing Yang Ya-Ping Chen Jun Liu Jing Zhao E Sun Xiao-Bin Jia Liang Feng State Key Laboratory of Natural Medicines School of Traditional Chinese Pharmacy China Pharmaceutical University College of Pharmacy Xuzhou Medical University Third Clinical Medical College Nanjing University of Chinese Medicine Jumpcan Pharmaceutical Co. Ltd.

Objective: Allergic asthma(AA) is a chronic airway infiammatory disease characterized by airway hyper-responsiveness(AHR). Pudilan anti-infiammatory oral liquid(PDL), along with its main medicinal material, Taraxaci Herba(Taraxacum mongolicum Hand.-Mazz, TH),has been widely used to treat upper respiratory tract infections. Research has shown that the major ingredient of TH, the organic acid component(OAC), possesses favorable AA activity. However, the attenuated effects of PDL and OAC from TH(TH-OAC) on AA and their possible mechanisms remain poorly understood. This study analyzed the attenuating effects of PDL and TH-OAC on AA and the underlying mechanisms. Methods: Young BALB/c mice were sensitized and stimulated to develop asthma using ovalbumin. Histological examinations were performed by hematoxylin and eosin staining. Western blotting, immunohistochemistry, and protein expression detection of toll-like receptor 2(TLR2), TLR4, and orosomucoid 1-like protein 3(ORMDL3) were performed to detect the presence of infiammatory components in the lung tissue. The messenger RNA(mRNA) expression levels were determined using quantitative real-time polymerase chain ***: Results showed that PDL and TH-OAC alleviated augmented AHR and typical asthmatic pathological changes, including infiammatory infiltration and thickening of the alveolar wall. They also significantly reduced the levels of the immunoglobulin E, IL-4, IL-5, IL-6, tumor necrosis factor-α, and Nitric oxide(NO) in lung tissues of mice. Protein and mRNA expression levels of TLR2, TLR4, and ORMDL3 were downregulated following treatment with PDL and TH-OAC. Conclusions: PDL and TH-OAC can reduce asthma-induced infiammatory damage to the bronchi. These results provide a theoretical basis for the treatment of asthma in clinical settings.

关键词： Allergic asthma organic acid component Pudilan anti-inflammatory oral liquid Taraxaci Herba toll-like receptor 2 toll-like receptor 4

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Role of toll like-receptor 2 in inflammatory activity of macrophage infected with a recombinant BCG expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum

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Asian Pacific Journal of Tropical Biomedicine 2018年第7期8卷 333-339页

作者： Nor Munirah Zakaria Rapeah Suppian Norazmi Mohd Nor Nor Fazila Che Mat School of Health Sciences Health Campus Universiti Sains Malaysia

Objective: To investigate the role of toll-like receptor 2(TLR2) in inflammatory activity of macrophage infected with the recombinant Mycobacterium bovis bacillus Calmette-Guerin(rBCG). Methods: Mouse macrophage cell line J774 A.1 was infected with Mycobacterium bovis bacillus Calmette-Guerin(BCG) and rBCG cultures for 48 h in the presence or absence of 10 μg/mL of TLR2 inhibitor. Untreated macrophages were used as a negative control while lipopolysaccharide-stimulated macrophages were used as a positive control. The ability of the macrophage to engulf the BCG and rBCG in the absence or presence of TLR2 inhibitor was assessed using a phagocytic assay, while the production of inflammatory cytokines and nitric oxide by the infected macrophages was evaluated using ELISA and Griess reagent method, while the expression of the inducible nitric oxide synthase was determined using Western blot analysis. Results: The results showed that blocking TLR2 function reduced the phagocytic activity, nitric oxide production and proinflammatory cytokine secretion such as TNF-α, IL-1β and IL-12 p40 as well as inducible nitric oxide synthase expression in the infected macrophages. These data showed the importance of TLR2 in the activation of macrophages following BCG and r BCG infections. Conclusions: Through exploring the immunological mechanism which underlies the protection conferred by the candidate vaccine, this study will improve our understanding of the vaccine candidate's mechanism to protect the host from malaria infection.

关键词： Macrophages Mycobacterium bovis bacillus Calmette-Guerin Plasmodium falciparum Proinflammatory cytokine toll-like receptor 2

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Argon preconditioning protects neuronal cells with a toll-like receptor-mediated effect

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Neural Regeneration Research 2023年第6期18卷 1371-1377页

作者： Stefanie Scheid Adrien Lejarre Jakob Wollborn Hartmut Buerkle Ulrich Goebel Felix Ulbrich Department of Anesthesiology and Critical Care Medical Center-University of FreiburgFaculty of MedicineFreiburgGermany Department of Anesthesiology Perioperative and Pain MedicineBrigham and Women’s HospitalHarvard Medical SchoolBostonMAUSA Department of Anesthesiology and Critical Care Medicine St.Franziskus-HospitalMuensterGermany

The noble gas argon has the potential to protect neuronal cells from cell *** far,this effect has been studied in treatment after acute *** using argon has not yet been *** this study,human neuroblastoma SH-SY5Y cells were treated with different concentrations of argon(25%,50%,and 74%;21%O_(2),5%CO_(2),balance nitrogen)at different time intervals before inflicting damage with rotenone(20μM,4 hours).Apoptosis was determined by flow cytometry after annexin V and propidium iodide *** expressions of toll-like receptors 2 and 4 were also *** were also processed for analysis by western blot and qPCR to determine the expression of apoptotic and inflammatory proteins,such as extracellular-signal regulated kinase(ERK1/2),nuclear transcription factor-κB(NF-κB),protein kinase B(Akt),caspase-3,Bax,Bcl-2,interleukin-8,and heat shock *** staining was performed for TLR2 and 4 and *** were also pretreated with OxPAPC,an antagonist of TLR2 and 4 to elucidate the molecular *** showed that argon preconditioning before rotenone application caused a dose-dependent but not a time-dependent reduction in the number of apoptotic *** with 74%argon for 2 hours was used for further experiments showing the most promising *** decreased the surface expression of TLR2 and 4,whereas OxPAPC treatment partially abolished the protective effect of *** increased phosphorylation of ERK1/2 but decreased NF-κB and *** inhibited mitochondrial apoptosis and the heat shock *** also suppressed the expression of the pro-inflammatory cytokine *** confirmed the alteration of TLRs and *** reversed the argon effect on ERK1/2,Bax,Bcl-2,caspase-3,and interleukin-8 expression,but not on NF-κB and the heat shock *** together,argon preconditioning protects against apoptosis of neuronal c

关键词： apoptosis inflammation interleukin-8 neuroprotection rotenone SH-SY5Y toll-like receptor 2 toll-like receptor 4

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Lipopolysaccharide upregulates the expression of toll-like receptor 4 in human vascular endothelial cells

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Chinese Medical Journal 2002年第2期115卷 286-289,158页

作者：杨清武朱佩芳王正国蒋建新第三军医大学大坪医院野战外科研究所四室重庆400042

OBJECTIVE: To investigate the expression of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA was extracted from ECV304 cells and isolated human umbilical vein endothelial cells (HUVECs) exposed to LPS, respectively. The quantification of TLR2 and TLR4 mRNA in HUVECs and EVC304 cells was carried out by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ECV304 cells and HUVECs were able to express TLR2 and TLR4 mRNA, but the expression levels of TLR4 appeared to be stronger than those of TLR2. LPS could upregulate the expression levels of TLR4 obviously, whereas it had no effect on the expression level of TLR2. CONCLUSIONS: Our data indicate that TLR4 may be the LPS signal transducer in endothelial cells and plays important roles in the cell activation of LPS. The ECV304 cell line is a better experimental model than isolated HUVECs in the research of endothelial cells.

关键词： Drosophila Proteins Cell Line Dose-Response Relationship, Drug Endothelium, Vascular Gene Expression Regulation Humans Lipopolysaccharides Membrane Glycoproteins RNA, Messenger receptors, Cell Surface Research Support, Non-U.S. Gov't Time Factors toll-like receptor 2 toll-like receptor 4 toll-like receptors Umbilical Veins Up-Regulation

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Efficacy of catgut embedding in Baihui(GV20)and Feishu(BL13)and Pishu(BL20)on lung tissue,brain tissue and blood related indexes in rats with allergic rhinitis of lung deficiency type

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Journal of Traditional Chinese Medicine 2023年第5期43卷 973-982页

作者： HUANG Heyin WANG Yichen ZHANG Qinxiu WU Xiaojuan WANG Jiang Li Xiangyu PANG Rui FAN Li CAO Liu TANG Luo Department of ENT Sichuan Integrative Medicine HospitalChengdu 610041China Department of orthopaedics Sichuan Province Orthopaedic HospitalChengdu 610000China Department of ENT Affiliated Hospital of Chengdu University of Traditional Chinese MedicineChengdu 610000China Department of ENT Quxian Traditional Chinese Medicine HospitalQuxian 635299China

OBJECTIVE:To investigate the effects of acupoint catgut embedding for 3 weeks on lung tissue,blood immunoglobulin E(IgE)and interleutin-4(IL-4),brain tissue microglia x-42(OX-42)and toll-like receptor-2(TLR-2)in rats with allergic rhinitis of lung deficiency ***:Forty-five female Sprague-Dawley rats were randomly divided for two *** first time,they were randomly divided into model group and blank group(Group C)according to 2∶1,and the second time,the model group were randomly divided into model control group(Group B)and intervention treatment group(Group A)according to 1∶1.15 in each *** Group A and Group B,the lung deficiency model was made by"sulfur–moxa fumigation",and then the allergic rhinitis model was established by"ovalbumin(OVA)sensitization".Then catgut embedding was performed at acupoints in Group A and not in Group *** 3 weeks,collect lung tissue samples for hematoxylin-eosin staining,then take blood to observe the concentration of IgE and IL-4,and finally take brain tissue to observe the results of OX-42 and ***:IgE level(μg/m L)was(3.11±0.20)in the Group A,(4.19±0.44)in the Group B,and(2.29±0.30)in the Group C(all P<0.001).IL-4 level(pg/mL)was(14.2±0.7)in the Group A,(18.6±2.4)in the Group B,and(11.4±1.2)for the Group C(all P<0.001).The mean OD for OX-42 is(0.1728±0.0016)in the Group A,(0.1810±0.0046)in the Group B and(0.1674±0.0025)in the Group C(all P<0.001).CONCLUSION:Although 3 weeks of acupoint catgut embedding already showed obvious efficacy on rats with allergic rhinitis,the allergic reaction in the body still *** achieve further treatment,prolonging the catgut embedding time is necessary.

关键词： rhinitis,allergic acupoint catgut embedding enzyme-linked immunosorbent assay immunoglobulin E interleukin-4 hippocampus toll-like receptor 2

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