隐丹参酮通过TLR4/NF-κB/JNK信号通路减轻脂多糖对肺泡上皮细胞炎性损伤的作用研究

目的 本研究旨在探讨隐丹参酮(CTS)对脂多糖(LPS)诱导的肺泡上皮细胞(MLE-12)的影响。方法 通过CCK-8法检测细胞活力,酶联免疫吸附试验(ELISA)和实时荧光定量PCR(q-PCR)检测IL-1β、IL-6、TNF-α含量及基因表达水平,筛选出具有最佳造模效果的LPS浓度;之后通过CCK-8考察CTS对细胞的非毒性剂量范围;将试验分为对照组、LPS组和CTS+LPS组3个组,ELISA检测细胞上清中IL-1β、IL-6、TNF-α的含量,qPCR检测细胞内IL-1β、IL-6、TNF-α、Bax和Bcl-2基因表达水平,Western Blot检测Bax、Bcl-2、TLR4、p-p65和p-JNK蛋白水平。结果 与对照组相比,LPS浓度≥1.0μg/mL时细胞活力显著下降(P<0.01);2.0和5.0μg/mL LPS组IL-1β、IL-6和TNF-α含量及基因表达水平显著上升(P<0.05);CTS浓度为0~5.0μM时,细胞活力无显著变化(P>0.05),即药物的无毒范围。与LPS组相比,CTS+LPS组IL-1β、IL-6和TNF-α含量及基因表达水平显著下降(P<0.05);CTS+LPS组Bax蛋白及基因表达水平显著下降(P<0.01)、Bcl-2蛋白及基因表达水平显著上升(P<0.05)。另外,与对照组相比,LPS组TLR4、p-p65、p-JNK蛋白水平显著上升(P<0.01);与LPS组相比,CTS+LPS组TLR4、p-p65、p-JNK蛋白水平显著下降(P<0.01)。结论 CTS可能通过抑制TLR4/NF-κB/JNK信号通路减轻LPS诱导的MLE-12细胞炎性损伤作用。

Downregulation of MUC1 Inhibits Proliferation and Promotes Apoptosis by Inactivating NF-κB Signaling Pathway in Human Nasopharyngeal Carcinoma

Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital.The expression of MUC1 was measured by real-time quantitative PCR(qPCR)in the patients with PNC.The 5-8F and HNE1 cells were transfected with siRNA control(si-control)or siRNA targeting MUC1(si-MUC1).Cell proliferation was analyzed by cell counting kit-8 and colony formation assay,and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells.The qPCR and ELISA were executed to analyze the levels of TNF-αand IL-6.Western blot was performed to measure the expression of MUC1,NFкB and apoptosis-related proteins(Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues,and NPC patients with the high MUC1 expression were inclined to EBV infection,growth and metastasis of NPC.Loss of MUC1 restrained malignant features,including the proliferation and apoptosis,downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy,including cell proliferation and apoptosis,by inactivating NF-κB signaling pathway in NPC.

Apatinib reduces liver cancer cell multidrug resistance by modulating NF-κB signaling pathway

Objectives:This investigation aimed to elucidate the inhibitory impact of apatinib on the multidrug resistance of liver cancer both in vivo and in vitro.Methods:To establish a Hep3B/5-Fu resistant cell line,5-Fu concentrations were gradually increased in the culture media.Hep3B/5-Fu cells drug resistance and its alleviation by apatinib were confirmed via flow cytometry and Cell Counting Kit 8(CCK8)test.Further,Nuclear factor kappa B(NF-κB)siRNA was transfected into Hep3B/5-Fu cells to assess alterations in the expression of multidrug resistance(MDR)-related genes and proteins.Nude mice were injected with Hep3B/5-Fu cells to establish subcutaneous xenograft tumors and then categorized into 8 treatment groups.The treatments included oxaliplatin,5-Fu,and apatinib.In the tumor tissues,the expression of MDRrelated genes was elucidated via qRT-PCR,immunohistochemistry,and Western blot analyses.Results:The apatinibtreated mice indicated slower tumor growth with smaller size compared to the control group.Both the in vivo and in vitro investigations revealed that the apatinib-treated groups had reduced expression of MDR genes GST-pi,LRP,MDR1,and p-p65.Conclusions:Apatinib effectively suppresses MDR in human hepatic cancer cells by modulating the expression of genes related to MDR,potentially by suppressing the NF-κB signaling pathway.

Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates inflammatory response ulcerative colitis through TLR4/NF-κB signaling pathway

BACKGROUND Ulcer colitis(UC)is a chronic,nonspecific,and noninfectious inflammatory bowel disease.Recently,Toll-like receptors(TLRs)have been found to be closely associated with clinical inflammatory diseases.Achieving complete remission in patients with intermittent periods of activity followed by dormancy is challenging.Moreover,no study has explored the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.AIM To explore the mechanism by which Kuicolong-yu enema decoction retains traditional Chinese medicine enemas to attenuate the inflammatory response in UC.METHODS This prospective clinical study included patients who met the exclusion criteria in 2020 and 2021.The patients with UC were divided into two groups(control and experimental).The peripheral blood of the experimental and control groups were collected under aseptic conditions.The expression of TLR4 protein,NF-κB,IL-6,and IL-17 was detected in the peripheral blood of patients in the experimental group and control group before and 1 month after taking the drug.Linear co rrelation analysis was used to analyze the relationship between the expression level of TLR4 protein and the expression levels of downstream signal NF-κB and inflammatory factors IL-6 and IL-17,and P<0.05 was considered statistically significant.RESULTS There were no significant differences in the patient characteristics between the control and experimental groups.The results showed that the expression levels of TLR4 and NF-κB in the experimental group were significantly lower than those in the control group(P<0.05).The levels of IL-6 and IL-17 in the experimental group were significantly lower than those in the control group(P<0.05).The TLR4 protein expression in the experimental group was positively correlated with the expression level of downstream signal NF-κB and was positively correlated with the levels of downstream inflammatory cytokines IL-6 and IL-17(r=0.823,P<0.05).CONCLUSION Kuicolong-yu enema decoction retains traditional Chinese medicine enema attenuates the inflammatory response of UC through the TLR4/NF-κB signaling pathway.

Aszonapyrone A Isolated from Neosartorya spinosa IFM 47025 Inhibits the NF-κB Signaling Pathway Activated by Expression of the Ependymoma-Causing Fusion Protein ZFTA-RELA

Ependymoma is a rare and chemotherapy-resistant brain tumor, which has resulted in a delay in the development of drugs to treat it. A subclass of supratentorial ependymomas (ST-EPN), designated ST-EPN-zinc finger-translocation-associated (ZFTA, ST-EPN-ZFTA), exhibits the expression of a fusion protein comprising ZFTA and v-rel reticuloendotheliosis viral oncogene homolog A (RELA), an effector transcription factor of the nuclear factor-kappa B (NF-κB) pathway (ZFTA-RELA). The expression of ZFTA-RELA results in the hyperactivation of the oncogenic NF-κB signaling pathway, which ultimately leads to the development of ST-EPN-ZFTA. To identify inhibitors of the NF-κB signaling pathway activated by the expression of ZFTA-RELA, we used a doxycycline-inducible ZFTA-RELA-expressing NF-κB reporter cell line and found that extracts of the fungus Neosartorya spinosa IFM 47025 exhibited NF-κB inhibitory activity. We identified eight compounds [aszonapyrone A (2), sartorypyrone A (3), epiheveadride (4), acetylaszonalenin (5), (R)-benzodiazepinedione (6), aszonalenin (7), sartorypyrone E (8) and (Z, Z)-N,N’-(1,2-bis[(4-methoxyphenyl)methylene]-1,2-ethanediyl)bis-formamide (9)] from N. spinosa IFM 47025 culture extract using a variety of chromatographic techniques. The structures of these compounds were identified through the analysis of various instrumental data (1D, 2D-NMR, MS, and optical rotation). The NF-κB responsive reporter assay indicated that compounds 2, 3, 5, 7, and 9 exhibited inhibitory activity. We further evaluated the inhibitory activity of these compounds against the expression of endogenous NF-κB responsive genes (CCND1, L1CAM, ICAM1, and TNF) and found that compound 2 showed significant inhibitory activity. Further studies are required to elucidate the mechanism of action of compound 2, which may serve as a lead compound for the development of a novel therapy for ST-EPN-ZFTA.

Parthenolide enhances the metronomic chemotherapy effect of cyclophosphamide in lung cancer by inhibiting the NF-kB signaling pathway

BACKGROUND Parthenolide(PTL),a sesquiterpene lactone derived from the medicinal herb Chrysanthemum parthenium,exhibits various biological effects by targeting NF-kB,STAT3,and other pathways.It has emerged as a promising adjunct therapy for multiple malignancies.AIM To evaluate the in vitro and in vivo effect of PTL on cyclophosphamide(CTX)metronomic chemotherapy.METHODS The cytotoxicity of PTL and CTX on Lewis lung cancer cells(LLC cells)was assessed by measuring cell activity and apoptosis.The anti-tumor efficiency was evaluated using a tumor xenograft mice model,and the survival of mice and tumor volume were monitored.Additionally,the collected tumor tissues were analyzed for tumor microenvironment indicators and inflammatory factors.RESULTS In vitro,PTL demonstrated a synergistic effect with CTX in inhibiting the growth of LLC cells and promoting apoptosis.In vivo,metronomic chemotherapy com-bined with PTL and CTX improved the survival rate of tumor-bearing mice and reduced tumor growth rate.Furthermore,metronomic chemotherapy combined with PTL and CTX reduced NF-κB activation and improved the tumor immune microenvironment by decreasing tumor angiogenesis,reducing Transforming growth factorβ,andα-SMA positive cells.CONCLUSION PTL is an efficient compound that enhances the metronomic chemotherapy effects of CTX both in vitro and in vivo,suggesting its potential as a supplementary therapeutic strategy in metronomic chemotherapy to improve the chemotherapy effects.

白藜芦醇抑制海马组织NF-κB/JNK通路改善严重烧伤大鼠认知功能

目的探讨白藜芦醇(RSV)对严重烧伤大鼠认知功能的改善作用及其可能的机制。方法将雄性SD大鼠,随机分为对照组、模型组和RSV组,每组6只。建模成功后,RSV组大鼠每日灌胃20 mg/kg的RSV,对照组和模型组每日灌胃等体积的生理盐水溶液。4周后,跳台实验检测大鼠认知功能;ELISA检测大鼠血清中肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)含量;实时定量PCR和Western blot法分别检测海马组织中TNF-α、IL-6 mRNA和蛋白的表达;原位末端转移酶标记技术(TUNEL)检测海马神经细胞凋亡情况;Western blot法检测海马组织中核因子κB/c-Jun N-氨基末端激酶(NF-κB/JNK)通路相关蛋白表达情况。结果与模型组大鼠相比,RSV组大鼠认知功能提高,大鼠血清TNF-α和IL-6含量明显降低,海马组织中TNF-α、IL-6 mRNA及蛋白表达降低,海马神经元凋亡率降低,磷酸化的NF-κB p65/NF-κB p65(p-NF-κB p65/NF-κB p65)、p-JNK/JNK比值降低。结论RSV通过阻断NF-κB/JNK信号通路,抑制炎症反应和海马神经元凋亡,从而改善严重烧伤大鼠认知功能。

酒石酸布托啡诺基于JNK/NF-κB信号通路对大鼠脑缺血再灌注损伤的改善作用

目的探讨酒石酸布托啡诺对大鼠脑缺血再灌注损伤(CIRI)的改善作用,并分析相关作用机制。方法80只大鼠随机分为假手术组、模型组、酒石酸布托啡诺组、尼莫地平组,每组20只。采用Zea Longa法评估大鼠神经功能,检测大鼠脑含水量,TTC染色法检测大鼠脑梗死体积比,HE染色观察大鼠脑组织病理性改变,TUNEL法检测大鼠脑组织细胞凋亡,Western blot法检测炎性因子和通路相关蛋白表达。结果与假手术组比较,模型组大鼠存在明显脑组织病理损伤,大鼠神经功能评分、脑含水量、脑梗死体积比均增高,脑组织细胞凋亡水平和炎性因子肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)表达升高,磷酸化c-Jun氨基末端激酶(p-JNK)、核因子-κB(NF-κB)、核因子κB磷酸化65(p65)蛋白表达上调,差异均有统计学意义(P<0.05)。与模型组比较,酒石酸布托啡诺和尼莫地平能够有效改善CIRI大鼠脑组织病理损伤,降低神经功能评分,减少大鼠脑含水量和脑梗死体积比,并降低脑组织细胞凋亡水平和炎性因子表达,下调p-JNK、NF-κB、p65蛋白表达,差异均有统计学意义(P<0.05)。结论酒石酸布托啡诺能够通过调控JNK/NF-κB信号通路发挥抗炎和抗凋亡作用,改善大鼠脑缺血再灌注损伤。

Quercetin regulates depression-like behavior in CUMS rat models via TLR4/NF-κB signaling

Background:Depression is becoming increasingly prevalent around the world,imposing a substantial burden on individuals,families,as well as society.Quercetin is known to be highly effective in treating depression.However,additional research is needed to dissect the mechanisms of its anti-depressive effects.Methods:For this study,Sprague-Dawley(SD)rats were randomized into the control,model,quercetin,or fluoxetine group.The latter three groups were exposed to chronic unpredictable mild stress(CUMS)for 42 d.The first two groups received saline solution daily via oral gavage.Meanwhile,the quercetin group was orally administered a quercetin suspension(52.08 mg/kg)every day,while the fluoxetine group was orally administered a fluoxetine solution(2.08 mg/kg).Here,fluoxetine served as the positive control drug to compare the therapeutic effects of quercetin.The experimental period was 6 weeks.Depressive behaviors in rats were assessed through various physiological and behavioral measures.Additionally,pathological changes in hippocampal tissues were examined using Nissl staining.Serum cytokines were detected using an enzymelinked immunosorbent assay(ELISA),and immunohistochemistry was employed to quantify the levels and integral optical density(IOD)values of ionized calcium binding adaptor molecule-1(Iba-1)expression in the brain.Real-time fluorescence quantitative PCR(RT-qPCR)was utilized to evaluate the mRNA levels of inflammatory indicators as well as toll-like receptor 4(TLR4),and nuclear factor-κappa B P65(NF-κB P65)in hippocampus.Western blot(WB)technique was employed to observe the protein levels of TLR4,NF-κB P65,and phospho-NF-κB P65(p-NF-κB P65).Results:After 42 d of exposure to CUMS,rats exhibited a slow increase in body weight,a reduction in food intake,an abnormal preference for sugar water,and aberrant open-field behaviors.Pathological analysis revealed the disintegration,rupture,interruption,and disorganization of hippocampal neuronal cells after CUMS exposure,along with a decrease in Nissl bodies in the CA1 region.This was accompanied by the elevated expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in the serum and the upregulation of IL-1β,IL-6,and TNF-αmRNA expression in the hippocampus.Increases in Iba-1-positive cells and the IOD values of Iba-1 were detected in hippocampal microglia.Furthermore,TLR4 and NF-κB P65 mRNA and protein levels were upregulated in hippocampal tissues.Quercetin,an antidepressant,could alleviate depression-like symptoms in rats and downregulate inflammatory factors associated with the TLR4/NF-κB signaling pathway in hippocampal microglia,and its therapeutic effect was comparable to fluoxetine.Conclusion:In rat models of CUMS,quercetin may act as an antidepressant by inhibiting inflammation in hippocampal microglia via TLR4/NF-κB signaling pathway.These results offer experimental and theoretical support for applying quercetin in the clinical management of depression.

Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

BACKGROUND Hepatic fibrosis is a serious condition,and the development of hepatic fibrosis can lead to a series of complications.However,the pathogenesis of hepatic fibrosis remains unclear,and effective therapy options are still lacking.Our group identified hepatitis C virus nonstructural protein 3-transactivated protein 1(NS3TP1) by suppressive subtractive hybridization and bioinformatics analysis,but its role in diseases including hepatic fibrosis remains undefined.Therefore,additional studies on the function of NS3TP1 in hepatic fibrosis are urgently needed to provide new targets for treatment.AIM To elucidate the mechanism of NS3TP1 in hepatic fibrosis and the regulatory effects of calcitriol on NS3TP1.METHODS Twenty-four male C57BL/6 mice were randomized and separated into three groups,comprising the normal,fibrosis,and calcitriol treatment groups,and liver fibrosis was modeled by carbon tetrachloride(CCl4).To evaluate the level of hepatic fibrosis in every group,serological and pathological examinations of the liver were conducted.TGF-β1 was administered to boost the in vitro cultivation of LX-2 cells.NS3TP1,α-smooth muscle actin(α-SMA),collagen I,and collagen Ⅲ in every group were examined using a Western blot and real-time quantitative polymerase chain reaction.The activity of the transforming growth factor beta 1(TGFβ1)/Smad3 and NF-κB signaling pathways in each group of cells transfected with pcDNA-NS3TP1 or siRNA-NS3TP1 was detected.The statistical analysis of the data was performed using the Student’s t test.RESULTS NS3TP1 promoted the activation,proliferation,and differentiation of hepatic stellate cells(HSCs)and enhanced hepatic fibrosis via the TGFβ1/Smad3 and NF-κB signaling pathways,as evidenced by the presence of α-SMA,collagen I,collagen Ⅲ,p-smad3,and p-p65 in LX-2 cells,which were upregulated after NS3TP1 overexpression and downregulated after NS3TP1 interference.The proliferation of HSCs was lowered after NS3TP1 interference and elevated after NS3TP1 overexpression,as shown by the luciferase assay.NS3TP1 inhibited the apoptosis of HSCs.Moreover,both Smad3 and p65 could bind to NS3TP1,and p65 increased the promoter activity of NS3TP1,while NS3TP1 increased the promoter activity of TGFβ1 receptor I,as indicated by coimmunoprecipitation and luciferase assay results.Both in vivo and in vitro,treatment with calcitriol dramatically reduced the expression of NS3TP1.Calcitriol therapy-controlled HSCs activation,proliferation,and differentiation and substantially suppressed CCl4-induced hepatic fibrosis in mice.Furthermore,calcitriol modulated the activities of the above signaling pathways via downregulation of NS3TP1.CONCLUSION Our results suggest that calcitriol may be employed as an adjuvant therapy for hepatic fibrosis and that NS3TP1 is a unique,prospective therapeutic target in hepatic fibrosis.

Acupuncture at Back-Shu point improves insomnia by reducing inflammation and inhibiting the ERK/NF-κB signaling pathway

BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use is prone to drug resistance and other adverse reactions.Acupuncture has a good curative effect and unique advantages in the treatment of insomnia.AIM To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia.METHODS We first prepared a rat model of insomnia,and then carried out acupuncture for 7 consecutive days.After treatment,the sleep time and general behavior of the rats were determined.The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats.The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA.qRTPCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway.Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1,MEK-2,ERK1/2 and NF-κB.RESULTS Acupuncture can prolong sleep duration,and improve mental state,activity,diet volume,learning ability and spatial memory.In addition,acupuncture increased the release of 1L-1β,1L-6 and TNF-αin serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway.CONCLUSION These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippocampus.

Huangqin decoction alleviates lipid metabolism disorders and insulin resistance in nonalcoholic fatty liver disease by triggering Sirt1/NF-κB pathway

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)is a clinicopathological entity characterized by intrahepatic ectopic steatosis.As a consequence of increased consumption of high-calorie diet and adoption of a sedentary lifestyle,the incidence of NAFLD has surpassed that of viral hepatitis,making it the most common cause of chronic liver disease globally.Huangqin decoction(HQD),a Chinese medicinal formulation that has been used clinically for thousands of years,has beneficial outcomes in patients with liver diseases,including NAFLD.However,the role and mechanism of action of HQD in lipid metabolism disorders and insulin resistance in NAFLD remain poorly understood.AIM To evaluate the ameliorative effects of HQD in NAFLD,with a focus on lipid metabolism and insulin resistance,and to elucidate the underlying mechanism of action.METHODS High-fat diet-induced NAFLD rats and palmitic acid(PA)-stimulated HepG2 cells were used to investigate the effects of HQD and identify its potential mechanism of action.Phytochemicals in HQD were analyzed by highperformance liquid chromatography(HPLC)to identify the key components.RESULTS Ten primary chemical components of HQD were identified by HPLC analysis.In vivo,HQD effectively prevented rats from gaining body and liver weight,improved the liver index,ameliorated hepatic histological aberrations,decreased transaminase and lipid profile disorders,and reduced the levels of pro-inflammatory factors and insulin resistance.In vitro studies revealed that HQD effectively alleviated PA-induced lipid accumulation,inflammation,and insulin resistance in HepG2 cells.In-depth investigation revealed that HQD triggers Sirt1/NF-κB pathwaymodulated lipogenesis and inflammation,contributing to its beneficial actions,which was further corroborated by the addition of the Sirt1 antagonist EX-527 that compromised the favorable effects of HQD.CONCLUSION In summary,our study confirmed that HQD mitigates lipid metabolism disorders and insulin resistance in NAFLD by triggering the Sirt1/NF-κB pathway.

Mechanism of Sanshi decoction inhibits macrophage pyroptosis by inhibiting BRD4/NF-κB/NLRP3 pathway in the treatment of gouty arthritis

Objective:To observe the effect of Sanshi decoction on BRD4/NF-κB/NLRP3 pathwaymediated macrophage pyroptosis,so as to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthritis.Methods:THP-1 was induced into macrophages with foboside and the divided into the control group,model group,low-dose,medium-dose,high-dose group of Sanshi decoction,and BRD4 inhibitor group.Except for the control group,the remaining groups were induced with monosodium urate crystals to construct a gouty arthritis cell model.The activity of macrophages was detected by CCK8,the level of macrophage pyroptosis was detected by flow cytometry,the activity of LDH,the content of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay,and the expression of related proteins in the BRD4/NF-κB/NLRP3 pathway was detected by Western blot.Results:Compared with the control group,macrophage activity was decreased in the model group,and the level of pyroptosis,LDH activity,contents of IL-1β and IL-18,expression levels of BRD4,p-NF-kB p65,NLRP3,Caspase-1 p20,and IL-1β protein were significantly up-regulated,the differences were statistically significant(P<0.05 and P<0.01).Compared with the model group,macrophage activity was up-regulated in the Sanshi Decoction,and the level of pyroptosis,LDH activity,IL-1β and IL-18 contents,expression levels of BRD4,p-NF-kB p65,NLRP3,Caspase-1 p20,and IL-1β protein were significantly decreased with statistically significant differences(P<0.05 and P<0.01).Conclusion:Sanshi decoction inhibits macrophage pyroptosis by inhibiting BRD4/NF-κB/NLRP3 pathway activation,thus improving the inflammation level of gouty arthritis.

川崎病中IgM-AECA通过JNK、p38及NF-κB诱导内皮细胞ICAM-1的表达

目的川崎病(Kawasaki Disease,KD)大部分病人血清中存在抗内皮细胞受体抗体-IgM(IgM An-ti-endothelial cell antibodies,IgM-AECA)。该研究旨在探讨抗内皮细胞受体抗体(AECA)通过何种途径诱导内皮细胞细胞内吸附分子-1(intercellular adhesion molecule-1,ICAM-1)的表达以激活血管内皮细胞。方法检测30名KD患儿血清IgM-AECA浓度,并从17名AECA阳性KD患儿血清中提纯IgM-AECA,用IgM-AECA直接刺激人冠状动脉内皮细胞(human coronary artery endothelial cells,HCAEC)后,通过R T-PCR及ELISA方法检测ICAM-1 mR NA及蛋白的表达,同时应用了PD98059,SB203580,dimethylaminopurine(DMAP)和parthenolide,4种蛋白激酶阻断剂观察IgM-AECA通过何种途径激活HCAEC。结果 KD病人血清中的IgM-AECA可使HCAEC的ICAM-1 mRNA及蛋白表达明显增加,SB203580(p38阻断剂)、DMAP(JNK阻断剂)及parthenolide(NF-κB阻断剂)明显抑制ICAM-1的表达(均P<0.01),而PD98059(ERK1/2阻断剂)不能抑制ICAM-1的表达(P>0.05)。结论 KD病人血清中的IgM-AECA通过增强ICAM-1的表达来激活内皮细胞,IgM-AECA通过p38、JNK MAPK及NF-κB通路诱导ICAM-1的表达,IgM-AECA可能在KD冠状动脉病变的发生中发挥重要作用。

NF-κB参与低氧预适应对自体原位肝移植大鼠JNK通路的影响

背景:在肝脏缺血再灌注损伤中,NF-κB与JNK通路的串扰方式决定了细胞的死亡或存活。而将低氧预适应用于肝移植过程所导致的细胞凋亡现象,尚未见有报道。目的:探讨低氧预适应后NF-κB的表达对移植肝JNK通路的影响及保护作用。方法:采用经门静脉灌注法建立大鼠自体原位肝移植模型,SD大鼠随机分为正常对照组:不接受任何处理;自体移植组:行自体原位肝移植;低氧预适应组:移植前体积分数8%氮氧混合气体的低氧预处理90min,然后行自体原位肝移植。分别于移植后1,6,24h处死大鼠取肝脏标本检测肝组织丙二醛、超氧化物歧化酶水平,免疫组化方法测定大鼠肝组织p-JNK蛋白,RT-PCR检测肝脏NF-κB mRNA含量,透射电子显微镜下观察肝细胞的超微结构变化。结果与结论:与对照组比较,两移植组肝组织丙二醛水平升高,超氧化物歧化酶水平降低(P<0.05);与自体移植组比较,低氧预适应组丙二醛水平显著降低、超氧化物歧化酶水平显著升高(P<0.05)。与正常对照组比较,两移植组各时相p-JNK蛋白的表达、NF-κB mRNA的转录水平显著升高(P<0.05);与自体移植组比较,低氧预适应组NF-κB mRNA转录水平显著增高(P<0.05),p-JNK蛋白的表达明显降低(P<0.05)。透射电镜下自体移植组肝细胞出现典型的凋亡征象,而低氧预适应组肝细胞无明显凋亡形态。提示,肝移植大鼠低氧预适应后可能通过上调NF-κB的表达,减少活性氧释放,抑制JNK通路的持续激活,从而抑制肝细胞凋亡,减轻肝脏缺血再灌注。

瑞香素抑制c-Jun氨基末端激酶/核因子κB(JNK/NF-κB)通路减轻大鼠心肌缺血损伤

目的研究瑞香素(DAP)对于心肌缺血损伤大鼠的保护作用及其作用机制。方法将75只SD大鼠随机分为空白组、模型组、(30、60、90)mg/kg DAP处理组。模型的建立则是采用皮下注射85 mg/kg异丙基肾上腺素(ISO)连续2 d建立大鼠心肌缺血模型,持续灌胃给予相应剂量DAP处理7 d,观察DAP对心肌损伤的保护作用。采用大鼠心电图测量装置测定各组实验大鼠的心电图变化,采用分光光度法测定血清超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)酶活性和丙二醛(MDA)含量,ELISA检测血清肌肉B型肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、磷酸肌酸激酶(CPK)水平以及心肌组织匀浆白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)含量的变化;采用HE染色以及Masson染色观察心肌细胞的损伤及纤维化情况,实时定量PCR检测心肌组织c-Jun氨基末端激酶(JNK)和核因子κB(NF-κB)的mRNA水平,Western blot法检测JNK、NF-κB的磷酸化情况。结果DAP能增加心肌组织SOD、CAT、GSH-Px酶活性,降低MDA,并能降低血清组织中CK-MB、LDH、CPK、TNF-α和IL-1β水平,减少心肌细胞纤维化,抑制JNK和NF-κBp65的表达和磷酸化。结论DAP通过抑制JNK/NF-κB信号通路活化,抑制心肌缺血诱发的心肌细胞凋亡和纤维化。

JNK3通过与RelA/p65相互作用抑制NF-κB信号通路减弱Bel-7402细胞的黏附能力(英文)

JNK信号通路在细胞的炎症、增殖与凋亡等生物学过程中发挥了重要的作用.我们采用酵母双杂交技术发现转录因子p65是JNK3的相互作用蛋白质.体内体外实验均证实JNK3与p65存在蛋白质相互作用.报告基因实验结果表明过表达JNK3抑制TNFα诱导NF-κB介导的转录激活.EMSA结果证明JNK3减弱NF-κB的DNA结合能力.实时定量PCR结果表明JNF3减少NF-κB靶基因的表达.综上所述,我们的研究结果表明JNK3做为一个调节分子在体内发挥了抑制p65转录活性的功能.

黄芪多糖通过抑制NF-κB和JNK信号通路减轻LPS诱导的小鼠心肌细胞凋亡

目的研究黄芪多糖(Astragalus polysaccharide,APS)对脂多糖(lipopolysaccharide,LPS)诱导的小鼠心肌细胞凋亡的影响,并探讨其作用机制。方法体外实验采用H9c2细胞预先给予APS,30 min后加入LPS(1 mg·L^(-1))共孵育24 h,建立心肌细胞凋亡模型。体内实验采用SPF级昆明小鼠预防性给予APS 14 d后,腹腔注射LPS(10 mg·kg^(-1))建立心肌细胞凋亡模型。8 h后采用超声心动测定小鼠心脏射血分数(EF)、左心室缩短分数(FS)等;TUNEL测心肌细胞凋亡;ELISA检测血清中IL^(-1)β、TNF-α含量;Western blot检测组织和体外心肌细胞中JNK、NF-κB信号通路及Bcl-2家族、caspase-3相关蛋白表达。结果 LPS能明显抑制小鼠的左心室收缩功能;促使心肌细胞凋亡;增加血清中IL^(-1)β、TNF-α,心肌细胞中JNK、p-JNK、Bax、caspase-3,胞核中NF-κB蛋白浓度;降低Bcl-2和胞质中NF-κB、IκB-α蛋白浓度。APS能明显保护LPS诱导的小鼠心肌收缩功能,减少心肌细胞凋亡;减少血清中IL^(-1)β、TNF-α,心肌组织细胞中p-JNK、Bax、caspase-3和胞核NF-κB蛋白含量;相对增加Bcl-2和胞质中NF-κB、IκB-α蛋白含量。而JNK蛋白表达无明显变化。结论 APS通过抑制NF-κB和JNK信号通路,减轻LPS诱导的小鼠心肌细胞凋亡。

度洛西汀通过调控JNK和NF-κB信号通路对急性心肌梗死大鼠心功能的影响

目的探究度洛西汀通过调控c-Jun氨基末端激酶(JNK)通路和核因子(NF)-κB信号通路对急性心肌梗死(AMI)大鼠心功能的影响。方法40只大鼠随机分为Sham组、AMI组、AMI+低剂量度洛西汀(AMI+L)组和AMI+高剂量度洛西汀(AMI+H)组(n=10)。通过结扎建立AMI模型,AMI+L组和AMI+H组大鼠使用度洛西汀灌胃,剂量分别为40 mg/kg和80 mg/kg。干预7 d后检测大鼠左室收缩末期压力(LVESP)、左室舒张末期压力(LVEDP)、心室收缩期间左心室压力的最大上升和下降速率(±dP/dt max)、心肌梗死面积、心肌细胞凋亡指数的影响。并通过ELISA检测各组细胞中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、超氧化歧化酶(SOD)、丙二醛(MDA),比较各组心肌组织中JNK和NF-κB mRNA和蛋白的水平。结果四组的上述指标比较差异显著(P<0.05)。AMI组的LVESP、±dP/dt max、SOD显著低于Sham组,LVEDP、梗死面积(2.41±0.39 mm^(2))、凋亡指数(32.75±3.67%)、IL-6、TNF-α、MDA、JNK和NF-κB mRNA和蛋白的水平显著高于Sham组(P<0.05)。AMI+L组的LVESP、±dP/dt max、SOD显著高于AMI组,LVEDP、梗死面积(2.02±0.42 mm^(2))、凋亡指数(21.12±2.74%)、IL-6、TNF-α、MDA、JNK和NF-κB mRNA和蛋白的水平显著低于AMI组(P<0.05)。AMI+H组的LVESP、±dP/dt max、SOD显著高于AMI组和AMI+L组,LVEDP、梗死面积(1.69±0.41 mm^(2))、凋亡指数(16.64±3.11%)、IL-6、TNF-α、MDA、JNK和NF-κB mRNA和蛋白的水平显著低于AMI组和AMI+L组(P<0.05)。结论度洛西汀可能通过抑制JNK和NF-κB通路抑制AMI大鼠心肌细胞凋亡,从而发挥保护心功能的作用。

Blautia producta displays potential probiotic properties against dextran sulfate sodium-induced colitis in mice

Blautia has attracted attention because of its potential efficacy in ameliorating host energy metabolism and inflammation.This study aims to investigate the influences of Blautia producta D4 on colitis induced by dextran sulfate sodium(DSS)and to reveal the underlying mechanisms.Results showed that B.producta D4 intervention significantly relieved body weight loss,and suppressed the elevation of pro-inflammatory cytokines(including interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β))and excessive oxidative stress(myeloperoxidease(MPO)activity,superoxide dismutase(SOD)activity,glutathione peroxidase(GSH-Px)activity,and malondialdehyde(MDA)level)in colitis mice.Moreover,the concentrations of tight junction proteins(occludin,claudin-1,and ZO-1)related to the intestinal barrier were obviously elevated,and colitis-related TLR4/NF-κB pathway activation was remarkably inhibited after B.producta D4 intervention.The intestinal microbial disorder was evidently ameliorated by increasing the relative abundance of Clostridium sensu stricto 1,Bifidobacterium,GCA-900066225,Enterorhabdus,and reducing the relative abundance of Lachnospiraceae NK4A136 group.In conclusion,oral administration of B.producta D4 could ameliorate DSS-induced colitis by suppressing inflammatory responses,maintaining the intestinal barrier,inhibiting TLR4/NF-κB pathway,and regulating intestinal microbiota balance.These results are conducive to accelerate the development of B.producta D4 as a functional probiotic for colitis.