Effects of Compound Fo'ercao Mixture on TLR4/MyD88/NF-κB signaling pathway in a rat model of chronic obstructive pulmonary disease / 中成药

AIM To explore the effect of Compound Fo'ercao Mixture on TLR4/MyD88/NF-κB signaling pathway in a rat model of chronic obstructive pulmonary disease(COPD).METHODS Rats were randomly divided into the blank group(n=10),and the model group(n=50)for the establishment of a rat model of COPD by 12-week cigarette smoke exposure combined with intratracheal injection of LPS.The successful rat models were randomly divided into the model group,the dexamethasone group(0.5 mg/kg)and the low,medium and high dose Compound Fo'ercao Mixture groups(6.8,13.6 and 27.2 g/kg),with 10 rats in each group.After 24 weeks of drug intervention,the rats had their lung function detected by animal lung function meter;their pathological changes of lung tissue observed by HE staining;their serum TNF-α,IL-1β,IL-6,and MDA levels and SOD activity detected by ELISA;their pulmonary mRNA expressions of TLR4,MyD88,NF-κB and caspase-3 detected by RT-qPCR;and their pulmonary protein expressions of TLR4,MyD88,NF-κB and TNF-α detected by Western blot.RESULTS Compared with the blank group,the model group displayed obviously pulmonary ventilation dysfunction,damaged lung tissue and bronchus,decreased SOD activity(P＜0.01);increased serum TNF-α,IL-1β,IL-6 and MDA levels(P＜0.01);and increased pulmonary expressions of TLR4,MyD88,NF-κB and caspase-3 mRNA and TLR4,MyD88,NF-κB and TNF-α proteins(P＜0.01).Compared with the model group,all Compound Fo'ercao Mixture groups shared improvement in lung function indices levels and lung tissue damage;decrease in the levels of serum TNF-α,IL-1β,IL-6 and MDA(P＜0.05,P＜0.01);and decrease in the pulmonary expressions of TLR4,MyD88,NF-κB and caspase-3 mRNA and TLR4,MyD88,NF-κB and TNF-α protein(P＜0.05,P＜0.01)in a dose-dependent manner.CONCLUSION Compound Fo'ercao Mixture can improve the lung dysfunction and pathological injury in a rat model of COPD,and its mechanism may be associated with the regulated TLR4/MyD88/NF-κB signaling pathway.

Inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia on transplanted tumor of colorectal cancer in nude mice / 中国药房

OBJECTIVE To study the inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia （TF-MPL） on transplanted tumor of colorectal cancer in nude mice. METHODS The transplanted tumor model of colorectal cancer was induced by injecting 0.2 mL colorectal cancer cell LoVo subcutaneously via the right armpit of nude mice. After successful modeling， nude mice were randomly divided into model group， 5-fluorouracil group （positive control， 10 mg/kg）， TF-MPL high- dose and low-dose groups （25， 12.5 mg/kg）； a normal group （normal saline containing 0.3% carboxymethyl cellulose sodium） without modeling was additionally set up， with 6 mice in each group. Each group was intraperitoneally injected with the corresponding drug solution/solvent for 21 consecutive days. The inhibitory rate of the transplanted tumor， liver and spleen index， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected after the last medication； the morphological changes of tumor tissue were observed； immunohistochemical staining was used to detect protein expressions of Toll- like receptor 4 （TLR4） and nuclear factor-κB subunit p65 （NF-κB p65） in tumor tissue of nude mice. Western blot assay was used to detect protein expressions of TLR4， myeloid differentiation factor 88 （MyD88）， TNF receptor-associated factor 6 （TRAF6）， interleukin-1 receptor-associated kinase 1 （IRAK-1）， NF-κB p65 and caspase-3 in tumor tissue of nude mice. RESULTS Compared with the model group， TF-MPL high-dose group showed a significant decrease in tumor weight （inhibitory rate of 36.91%）， liver and spleen index， serum levels of TNF-α and IL-6 and protein expressions of TLR4， MyD88， TRAF6，IRAK-1 and NF- κB p65 （P＜0.05 or P＜0.01）； the expression of caspase-3 protein was increased significantly （P＜0.05）， and more tumor cell shrinkage and deformation， nuclear pyknosis and fragmentation were observed. CONCLUSIONS TF-MPL can significantly inhibit the growth of transplanted tumor of colorectal cancer in nude mice， the mechanism of which may be associated with reducing inflammatory response， inhibiting TLR4/MyD88/NF-κB signaling pathway， and promoting apoptosis in colorectal cancer cells.

Effects of Wheat Grain Moxibustion on the Expression of TLR4/MyD88/NF-κB Signaling Pathway in Spinal Cord Tissue of Rats with Sciatic Nerve Injury / 中国中医药信息杂志

Objective To observe the effects of wheat grain moxibustion for"Huantiao"on sciatic nerve function,pathological morphology of sciatic nerve stem and expressions of TLR4/MyD88/NF-κB signaling pathway expression in spinal cord tissue of rats with sciatic nerve injury(SNI);To explore the possible mechanism of wheat grain moxibustion for the treatment of SNI.Methods Totally 24 SD male rats were randomly divided into blank group,sham-operation group,model group and wheat grain moxibustion group,with 6 rats in each group.The model group and the wheat grain moxibustion group used a rat model with sciatic nerve clamping injury.From the 7th day after modeling,the rats were treated with moxibustion on the affected side of"Huantiao"for 6 strokes each time,once a day,for consecutive 10 days.The sciatic nerve function index(SFI)of rats on the 7th day after modeling and after intervention were observed,mechanical withdraw threshold(MWT)in rats were measured using a fiber optic pain gauge,ELISA was used to detect NO and iNOS content in spinal cord tissue,HE staining was used to observe the morphology of sciatic nerve stem,the expression of TLR4,NF-κBp65,p-NF-κBp65,MyD88,IκBα and p-IκBα in spinal cord tissue were detected by Western blot.Results Compared with the sham-operation group,the SFI and MWT of the rats in the model group significantly decreased(P<0.01),the arrangement of nerve fibers in sciatic nerve stem was disordered,with a significant increase in the number of Schwann cells and a large number of vacuolar degeneration,the content of NO,iNOS and the expression of TLR4,p-NF-κBp65,MyD88,p-IκBα protein in spinal cord tissue significantly increased(P<0.01).Compared with the model group,the SFI and MWT of the rats in the wheat grain moxibustion group increased significantly(P<0.01),the damage of sciatic nerve stem was reduced,with orderly cell arrangement,a decrease in the number of Schwann cells,and a decrease in axonal demyelination and cellular vacuolar degeneration,the content of NO,iNOS and the expression of TLR4,p-NF-κBp65,MyD88,p-IκBα in spinal cord tissue significantly decreased(P<0.05).Conclusion Wheat grain moxibustion for"Huantiao"can down-regulate TLR4,p-NF-κBp65,MyD88 and p-IκBα protein expressions in spinal cord tissue of SNI rats,reduce the secretion of NO and iNOS,thereby relieve pain and damaged nerve tissue inflammation response.

BMSCs inhibit inflammation in mice with LPS-induced acute lung injury by regulating TLR4/MyD88/NF-κB signaling pathway / 安徽医科大学学报

Objective To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on the inflamma-tory response of lipopolysaccharide(LPS)induced acute lung injury(ALI)in mice.Methods 32 SPF KM mice,aged 4 weeks were randomly divided into four groups,control group,LPS group,dexamethasone treatment group(LPS+DEX)and BMSCs treatment group(LPS+BMSCs).The latter three groups were injected with LPS by tra-cheal puncture to establish mouse ALI model 24 h after modeling,BMSCs isolated from the femur of mice were in-jected into the caudal vein,and DEX were injected into caudal vein at the same time in LPS+DEX group for 3 consecutive days.On the 4th day after cell transplantation or 24 h after DEX injection,the survival quantity of mice was recorded,lung function was detected,and the wet/dry weight ratio(W/D)of lung was measured.Then in-flammatory cells in bronchoalveolar lavage fluid(BALF),lung pathological changes and serum inflammatory cyto-kines were collected.Green fluorescent protein(GFP)staining was used to observe the homing of BMSCs in lung tissues.The mRNA and protein expression of TLR4,MyD88 and NF-κB in lung tissues were detected by RT-PCR and Western blot assay respectively.Results Compared with the control group,LPS model group showed de-creased lung function,significantly increase in the W/D weight ratio of lung,inflammatory cytokines in serum and inflammatory cells in BALF,and severe damage in lung tissue.Compared with LPS group,LPS+DEX group and LPS+BMSCs group showed improved lung function,reduced lung tissue damage,significantly decrease in the W/D weight ratio of lung,inflammatory cytokines in serum and inflammatory cells in BALF.And the expression of TLR4-MyD88-NF-κB signaling pathway-related genes and proteins decreased,the survival quantity increased.Conclusion Homologous BMSCs transplantation can effectively treat LPS-induced acute lung injury,and the mechanism may be related to the regulation of TLR4-MyD88-NF-κB signaling pathway and the reduction of inflam-matory response.These findings provide the experimental basis for BMSCs homologous transplantation for ALI.

Acupuncture Improve Scognitive Function of Vascular Dementia Rats by Regulating TLR-4/MyD88/NF-κB Pathway / 世界科学技术-中医药现代化

Objective To study the effect of acupuncture on the effect of acupuncture on the learning and memory ability and inflammatory factor expression in rats with vascular dementia,to provides an experimental basis for acupuncture and treatment of vascular dementia.Methods Randomly dividing thirty-six SPF grade SD male rats into 12 rats in sham surgery(A),model(B),and acupuncture(C).Groups B and C will prepare the VD model,and Group A will only surgically isolated the bilateral common carotid arteries without ligation.In Group C,acupuncture with"Baihui","Shenshu"and"Fenglong"points were used for intervention,each course lasts for 6 days,for a total of 2 sessions;Group A and Group B were fed normally without intervention.Morris water maze experiment was used to assess spatial learning and memory,ELISA for serum IL-1β and TNF-α,Real-time PCR for TLR-4,MyD88 and NF-κB mRNA,positive expression of Iba1 and NF-κB,and Western blot for relative expression levels of IL-1β and TNF-α in hippocampus.Results Compared with group B,group C rats had a shorter escape latency period(P<0.01)and had crossed the platform more often(P<0.05);The serum concentrations of TNF-α and IL-1β were significantly decreased(P<0.01);The expression amount of mRNA of TLR-4/MyD88/NF-κB was significantly reduced in the hippocampus(P<0.01);The positive expression of Iba1 and NF-κB protein in the brain was significantly reduced(P<0.01);The relative expression level of TNF-α and IL-1β in the hippocampus was significantly reduced(P<0.01).Conclusion Needle"Baihui","Shenshu"and"Fenglong"can improve vascular dementia rats cognitive dysfunction,and the mechanism may be related to the inhibition of neuroinflammatory response by inhibiting TLR-4/MyD88/NF-κB signaling pathway activity.

Effects of Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination on inflammatory responses in atherosclerotic mice / 中国中药杂志

The study aims to observe the effects and explore the mechanisms of Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination in the treatment of the inflammatory response of mice with atherosclerosis(AS) via the Toll-like receptor 4(TLR4)/myeloid differentiation primary response protein 88(MyD88)/nuclear factor-κB(NF-κB) signaling pathway. Male ApoE~(-/-) mice were randomly assigned into a model group, a Buyang Huanwu Decoction group, an Astragali Radix-Angelicae Sinensis Radix combination group, and an atorvastatin group, and male C57BL/6J mice of the same weeks old were used as the control group. Other groups except the control group were given high-fat diets for 12 weeks to establish the AS model, and drugs were administrated by gavage. Aortic intimal hyperplasia thickness, blood lipid level, plasma inflammatory cytokine levels, M1/M2 macrophage markers, and expression levels of proteins in TLR4/MyD88/NF-κB pathway in the vessel wall were measured to evaluate the effects of drugs on AS lesions and inflammatory responses. The results showed that the AS model was successfully established with the ApoE~(-/-) mice fed with high-fat diets. Compared with the control group, the model group showed elevated plasma total cholesterol(TC), triglyceride(TG), and low-density lipoprotein cholesterol(LDL-c) levels(P<0.05), thickened intima(P<0.01), and increased plasma tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels(P<0.01). Moreover, the model group showed increased expression of vascular cell adhesion molecule-1(VCAM-1) and inducible nitric oxide synthase(iNOS)(P<0.01), inhibited expression of endothelial nitric oxide synthase(eNOS) and cluster of differentiation 206(CD206)(P<0.01), and up-regulated mRNA and protein levels of TLR4, MyD88, NF-κB inhibitor alpha(IκBα), and NF-κB in the vessel wall(P<0.05). Compared with the model group, Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination lowered the plasma TC and LDL-c levels(P<0.01), alleviated the intimal hyperplasia(P<0.01), and reduced the plasma TNF-α and IL-6 levels(P<0.05). Moreover, the two interventions promoted the expression of eNOS and CD206(P<0.05), inhibited the expression of VCAM-1 and iNOS(P<0.01), and down-regulated the mRNA and protein levels of TLR4, MyD88, IκBα, and NF-κB(P<0.05) in the vessel wall. This study indicated that Buyang Huanwu Decoction and Astragali Radix-Angelicae Sinensis Radix combination could delay the progression of AS, inhibit the polarization of vascular wall macrophages toward M1 type, and attenuate vascular inflammatory response by inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway in the vascular wall. Astragali Radix and Angelicae Sinensis Radix were the main pharmacological substances in Buyang Huanwu Decoction for alleviating the AS vascular inflammatory response.

Anti-inflammatory effects of Abelmoschus manihot (L.) Medik. on LPS-induced cystitis in mice: potential candidate for cystitis treatment based on classic use / 中国天然药物

Abelmoschus manihot (L.) Medik. (A. manihot) is a traditional Chinese herbal medicine with a variety of pharmacological properties. It was first recorded in Jiayou Materia Medica dating back to the Song dynasty to eliminate urinary tract irritation by clearing away heat and diuretic effect. However, its pharmacological action on urinary tract infections has not been investigated. The present study aims to evaluate the anti-inflammatory activity of A. manihot on a mouse model of lipopolysaccharide (LPS)-induced cystitis. The results showed that A. manihot decreased white blood cell (WBC) count in urine sediments of the cystitis mice, alleviated bladder congestion, edema, as well as histopathological damage, reduced the expression levels of tumor necrosis factor-α, interleukin-6, and interleukin-1β simultaneously. Moreover, A. manihot administration significantly downregulated the expression levels of TLR4, MYD88, IκBα, p-IκBα, NF-κB p65, and p-NF-κB p65 in LPS-induced cystitis mice. These findings demonstrated the protective effect of A. manihot against LPS-induced cystitis, which is attributed to its anti-inflammatory profile by suppressing TLR4/MYD88/NF-κB pathways. Our results suggest that A. manihot could be a potential candidate for cystitis treatment.

Puerarin protects against myocardial ischemia/reperfusion injury by suppressing NLRP3 inflammasome activation via TLR4/Myd88/NF-κB pathway in rats / 药学学报

This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-κB) pathway were determined by Western blot. All animal experimental procedures were approved by the ethics committee of the Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical Sciences. The results showed that puerarin could significantly improve cardiac function, reduce myocardial infarct size, decease the levels of lactic dehydrogenase (LDH), aspartate transaminase (AST), creatine kinase-MB (CK-MB), and cardiac troponin T (cTnT) and suppress cardiomyocyte apoptosis. Meanwhile, puerarin could notably decrease the levels of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed that puerarin could downregulate the expression of TLR4, Myd88, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-caspase 1, cleaved-gasdermin-D (GSDMD), IL-1β, and IL-18, as well as the phosphorylation levels of inhibitor of NF-κB α (IκBα), IκB kinase β (IKKβ), and NF-κB. These findings demonstrated that puerarin could alleviate MI/R injury by suppressing NLRP3 inflammasome activation, possibly via TLR4/Myd88/NF-κB pathway.

Effect of lycopene on glycolipid metabolism and pancreatic tissue through regulation of TLR4/MyD88/NF-κB signaling pathway in high-fat diet induced obese mice / 中草药

Objective: To investigate the effects of lycopene on glycolipid metabolism and pancreatic tissue inflammation in obese mice and its underlying mechanism. Methods: The obese mouse model was induced by a high-fat diet (HFD). The effects of lycopene on body weight, blood glucose, blood lipid and body fat were observed after 8 weeks of administration. Pathological changes of pancreas were observed by HE staining. Protein expressions involved in TLR4/MyD88/NF-κB signaling pathway were detected by Western blotting, and the degree of macrophage infiltration in the pancreatic tissues of obese mice were detected by IHC. Results: Lycopene significantly inhibited body weight gain and reduced fasting blood glucose, as well as improved glucose tolerance and blood lipid level in obese mice. In addition, lycopene also reduced vacuolization, edema degeneration, islet hypertrophy and other inflammatory lesions in pancreatic tissues. Moreover, the protein expression levels of TLR4, MyD88 and NF-κB in pancreatic tissue were decreased and the inflammatory infiltration was reduced. Conclusion: Lycopene can improve blood glucose, lipid metabolism and pancreatic inflammation in obese mice, and its mechanism may be related to the regulation of TLR4/MyD88/NF-κB signaling pathway.

Intervention Effects of Berberine on Mice Macrophage Polarization Based on TLR 4/MyD88/NF-κB Signaling Pathway / 中国药房

OBJECTIVE：To study the effects of berberine on mic e macrophage polarization based on TLR 4-MyD88-NF-κB signaling pathway. METHODS ：Using mice RAW 264.7 macrophage as the object ，atorvastatin calcium as positive control ， inflammatory cell model was induced by lipopolysaccharide （LPS）；ELISA method was used to detect the contents of TNF-α，IL-6 and NF-κB in cell culture medium after treated with low，medium and high doses of berberine （5，10，20 μmol/L）for 24 h. The real-time fluorescence quantitative PCR was conducted to determine the mRNA expression of TLR 4 and MyD 88 in cells. Western blotting assay was used to detect the protein expression of TLR 4，MyD88，iNOS and CD 206 in cells. RESULTS ：Compared with blank control group ，the contents of TNF-α，IL-6 and NF-κB in cell culture medium，mRNA expression of TLR 4 and MyD 88， protein expression of TLR 4，MyD88 and iNOS in cells were increased significantly in LPS induction group （P＜0.05）. Compared with LPS induction group ，the contents of TNF-α and IL-6，mRNA and protein expression of TLR 4 and MyD 88 in atorvastatin calcium group ，berberine medium-dose and high-dose groupsas well as the content of NF-κ B and protein expression of iNOS in administration groups were decreased significantly ， while the content of NF-κB in berberine high-dose group was significantly lower than atorvastatin calcium group （P＜0.05）. The protein expressions of CD206 in atorvastatin calcium group and berberine high-dose group were increased significantly ，while the protein expression of CD 206 in berberine high-dose group was significantly higher than atorvastatin calcium group （P＜0.05）. CONCLUSIONS ：Different doses of berberine can intervene in mice macrophage polarization to different extents ，the mechanism of which may be associated with the regulation of TLR4/MyD88/NF-κB signaling pathway.

Regulatory effect of Di'ao Xinxuekang on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats / 中国中药杂志

The aim of this paper was to observe the effect of Di'ao Xinxuekang(DXXK) on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats, and to explore its anti-atherosclerotic mechanism. Sixty SD rats were randomly divided into normal group, model group, atorvastatin group(4.0 mg·kg~(-1)), and DXXK groups(100, 30, 10 mg·kg~(-1)), with 10 rats in each group. The atherosclerosis model was induced by high fat diet plus vitamin D_2. Experimental drugs were administered intragastrically once daily for 8 weeks starting from the 9 th week. Biochemical analyzers were used to detect levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in blood lipid. The levels of serum tumor necrosis factor(TNF)-α, interleukin(IL)-6 and IL-1β were detected by ELISA. Pathological changes of aortic tissues were observed by using Sudan Ⅳ and HE staining. The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 in aortic tissues were detected by RT-PCR and Western blot, respectively. As compared with the model group, TC, TG, and LDL-C levels in serum were significantly decreased, HDL-C content was significantly increased, and levels of TNF-α, IL-6, and IL-1β in serum were significantly decreased in atorvastatin group and DXXK high and middle dose groups. Aortic lesions in atorvastatin group and DXXK group were significantly improved, and the mRNA and protein expressions of TLR4, MyD88, NF-κB p65 in the aorta were decreased. DXXK has a preventive and therapeutic effect on atherosclerosis in rats, and its mechanism may be related to inhibiting inflammatory reaction by regulating TLR4/MyD88/NF-κB signal transduction, thereby inhibiting the progression of atherosclerosis.

Experimental study on effect and mechanism of Danzhi Jiangtang Capsules on diabetic myocardial injury / 中国中药杂志

Diabetic cardiomyopathy( DCM) is one of the major cardiovascular complications of diabetes mellitus. Based on the clinical efficacy of Danzhi Jiangtang Capsules( DJC) in the prevention and treatment of diabetes and its cardiovascular complications,both in vivo and in vitro methods were adopted to investigate its effect and underlying mechanism of protecting myocardial injury induced by diabetes. The type 2 diabetic rats were prepared by feeding high-energy food combined with streptozotin( STZ) injection,and the effects of DJC were observed by blood sugar,blood lipid,hemodynamic index,cardiac weight index and the change of cardiac pathological morphology. The protein expressions of TLR4,MyD88 and NF-κB p65 in myocardial tissue were detected and the possible mechanism was preliminarily analyzed. Besides this,DJC containing serum was prepared,H9 c2 cardiomyocyte induced by high sugar were studied to investigate the mechanism of TLR4/MyD88/NF-κB signaling pathway regulating cardiomyocyte injury and the therapeutic effect of DJC. The results demonstrated that fasting blood sugar,glycosylated hemoglobin,total cholesterol and glycerol triglyceride were significantly reduced( P<0. 01,P<0. 05). Cardiac weight index,left ventricle weight index,LVEDP and the protein expressions of TLR4,MyD88 and NF-κB p65 were significantly reduced( P<0. 01,P<0. 05). LVSP,+dp/dtmaxand-dp/dtmaxincreased significantly( P<0. 01,P< 0. 05). Moreover,the pathological damage of myocardial tissue in rats improved significantly. Meanwhile,the protein expressions of TLR4,MyD88 and NF-κB p65 in cardiomyocytes induced by high sugar were significantly inhibited( P<0. 01).It showed that DJC were effective in preventing and treating myocardial injury induced by diabetes and its mechanism may be related to the over-expression of TLR4/MyD88/NF-κB signaling pathway induced by high sugar.

Effect of Electroacupuncture Preconditioning on Myocardial Ischemia and Expression of TLR 4, MyD 88 and NF-κB mRNAs in "Neiguan" (PC 6) Area in Rats with Myocardial Ischemia-reperfusion Injury / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) preconditioning on acute myocardial ischemia (MI) and expression of Toll-like receptor 4 (TLR 4), myeloid differentiation factor 88 (MyD 88) and nuclear factor-κB (NF-κB) mRNAs in the "Neiguan" (PC 6) area of rats with MI-reperfusion injury (MIRI), so as to explore its mechanisms underlying improvement of MIRI. METHODS: Forty-eight male Wistar rats were randomly divided into control, control ＋ EA, sham operation, model, EA and sham EA groups (n＝8). The MIRI model was established by occlusion of the descending anterior branch of the left coronary artery for 30 min, followed by reperfusion. Before modeling, EA preconditioning was respectively applied to PC 6 or sham PC 6 for 30 min, once daily for 5 days. Electrocardiogram (ECG) of the standard limb-lead Ⅱ was recorded before and after the modeling, and 0.25 h after reperfusion. The expression levels of TLR 4, MyD 88 and NF-κB genes in the PC 6 tissue were detected by Real-time PCR. RESULTS: Compared with the control group, the height of ECG J-point was significantly increased in the model group (P0.05)．. CONCLUSION: EA preconditioning can improve myocardial ischemia and reduce the expression of TLR 4, MyD 88 and NF-κB mRNAs in PC 6 area of MIRI rats, suggesting an involvement of the regional cutaneous TLR 4/MyD 88/NF-κB signaling suppression in the initiation-transfer-amplification of EA stimulating signal inputs for relieving MIRI.

Tricin's inhibitory effects on TLR4/MyD88/NF-κB pathway of alveolar macrophages in asthma mice / 中成药

AIM To observe the inhibitory effects of tricin on the airway inflammation in low concentration LPS induced asthma mice.METHODS The model of asthma mice with airway inflammation was induced by low concentration LPS plus OVA.There were four groups:blank control group,asthma model group,tricin treatment group and dexamethasone (positive drug) treatment group.The levels of inflammatory factors,including NO,TNF-α and IL-1β,in bronchoalveolar lavage fluid (BALF) were detected by ELISA;the pathology of lung tissue was observed by HE staining;the mRNA and protein expressions of TLR4,MyD88 and NF-κB p65 in alveolar macrophages were detected using real-time quantitative PCR (RT-PCR) and Western blot.RESULTS Tricin could significantly decrease the levels of NO,TNF-α and IL-1β in BALF of asthma mice and improve lung inflammation.It could also down-regulate the relative mRNA and protein expressions of TLR4,MyD88 and NF-κB p65.CONCLUSION Tricin has inhibitory effects on airway inflammation in asthma mice,whose mechanism may be related to the intervention of TLR4/MyD88/NF-κB pathway.

Effects of Anzi Mixture on TLR4/MyD88/NF-κB Signaling Pathway of Antiphospholipid Antibodies Posi-tive Abortive Mice / 中国药房

OBJECTIVE:To study the effects of Anzi mixture on Toll like receptor 4(TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear facter-κB(NF-κB)signaling pathway of antiphospholipid antibodies(APA)positive abortive mice,and to inves-tigate the mechanism of anti-APA positive abortion. METHODS:BALB/c mice(female)were randomly divided into blank control group,model group,aspirin group (positive control,0.0195 g/kg) and Anzi mixture low-dose,medium-dose and high-dose groups (37.7,75.4,150.8 g/kg,calculated by crude drug),with 10 mice in each group. Except for blank control group,other groups were given human β2-glycoprotein Ⅰ as derivant to establish APA positive abortion model. From the first day of pregnancy, treatment groups were given relevant medicine intragastrically,and blank control group and model group were given constant vol-ume of normal saline intragastrically,once a day,for consecutive 9 d. mRNA and protein levels of TLR4,myeloid differentiation 2 (MD2),MyD88 and NF-κB in placental tissue of mice were determined by RT-PCR and immunohistochemical method. RE-SULTS:Compared with blank control group,mRNA and protein expression of TLR4,MD2,MyD88 and NF-κB in placental tis-sue were increased markedly in the model group(P<0.01). Compared with model group,mRNA and protein expression of TLR4, MD2 and MyD88 in aspirin group and Anzi mixture low-dose and medium-dose groups were decreased significantly as well as the protein expression of TLR4 in Anzi mixture high-dose group and the protein expression of NF-κB in all medicine groups(P<0.05 or P<0.01). mRNA expression of TLR4 and MD2 and the protein expression of MD2 and MyD88 in Anzi mixture low-dose groups were lower than those in aspirin group (P<0.05 or P<0.01). CONCLUSIONS:Anzi mixture can inhibit TLR4/MyD88/NF-κB signaling pathway of APA positive abortive mice,which may be one of anti-APA positive abortion mechanisms.