目的研究二甲双胍(metformin,Met)通过抑制NF-κBNLRP3通路改善草酸钙(Calcium Oxalate,CaOx)诱导人肾小管上皮细胞(HK-2)损伤。方法分别随机将人肾小管上皮细胞分成对照组、实验组(CaOx+HK-2)、高浓度Met组(CaOx+HK-2+1.2 mM Met)、低...目的研究二甲双胍(metformin,Met)通过抑制NF-κBNLRP3通路改善草酸钙(Calcium Oxalate,CaOx)诱导人肾小管上皮细胞(HK-2)损伤。方法分别随机将人肾小管上皮细胞分成对照组、实验组(CaOx+HK-2)、高浓度Met组(CaOx+HK-2+1.2 mM Met)、低浓度Met组(CaOx+HK-2+0.80 mM Met)、通路干预组(CaOx+HK-2+PDTC)五组。各组细胞培养24 h,采用酶联免疫吸附测定(Elisa)法检测细胞上清液中炎症因子(IL-6、IL-18、IL-1β),采用逆转录-聚合酶链反应(RT-PCR)法检测细胞中NF-κB、NLRP3、骨桥蛋白(OPN)mRNA。结果各组间细胞上清液IL-6、IL-18、IL-1β含量相比差异均有统计学意义(P均<0.05);各组间细胞中NF-κBmRNA表达量、NLRP3mRNA表达量、OPNmRNA表达量相比差异均有统计学意义(P均<0.05)。结论1.20 mmol/L或0.80 mmol/L Met可以通过抑制NF-κB/NLRP3通路调控下游的炎症相关蛋白及黏附蛋白表达,继而改善CaOx诱导人肾小管上皮细胞损伤及减少草酸钙肾结石形成。展开更多
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.Achievin...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.展开更多
目的探究高迁移率族蛋白1(High Mobility Group Box Protein1, HMGB1)通过(Toll-like Receptor4,TLR4)/核因子κB(Nuclear Factor Kappa-B, NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NOD-like Receptor Thermal Protein Domain Associat...目的探究高迁移率族蛋白1(High Mobility Group Box Protein1, HMGB1)通过(Toll-like Receptor4,TLR4)/核因子κB(Nuclear Factor Kappa-B, NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NOD-like Receptor Thermal Protein Domain Associated Protein 3, NLRP3)信号通路介导内皮细胞焦亡在系统性血管炎中的作用机制。方法 研究于2021年10月—2023年9月在齐齐哈尔医学院附属第三医院开展。取人脐静脉血管内皮细胞进行培养,随机分为对照组和实验组,实验组加入人重组HMGB1,对比对照组与实验组、实验组NLRP3及TLR4表达抑制前后,内皮细胞焦亡相关蛋白的表达水平。结果 系统性血管炎组与健康对照组比较,人脐静脉血管内皮细胞中HMGB1、含半胱氨酸的天冬氨酸蛋白水解酶-1(caspase-1)、白细胞介素-1β(interleukin-1β, IL-1β)、白细胞介素-18(interleukin-18, IL-18)水平升高,差异有统计学意义(P均<0.05)。细胞实验中实验组与对照组比较,caspase-1、IL-1β、IL-18水平升高,差异有统计学意义(P均<0.05)。过表达HMGB1并抑制NLRP3可使NLRP3(2.71±0.59 vs 1.24±0.58)、caspase-1(0.69±0.12 vs 0.40±0.03)、IL-1β[(1.75±0.31)pg/mL vs (1.16±0.12)pg/mL]、IL-18[(0.15±0.04)pg/mL vs (0.09±0.01)pg/mL]水平降低,差异有统计学意义(P均<0.05);过表达HMGB1并抑制TLR4可使TLR4(4.93±1.04 vs 1.96±0.84)、NF-κB(5.62±1.39 vs 2.15±1.04)、NLRP3(2.71±0.59 vs 1.24±0.58)、caspase-1(0.69±0.12 vs 0.40±0.03)、IL-1β[(1.75±0.31)pg/mL vs (1.16±0.12)pg/mL]、IL-18[(0.15±0.04)pg/mL vs (0.09±0.01)pg/mL]水平明显降低,差异有统计学意义(P均<0.05)。结论 HMGB1通过调节TLR4/NF-κB/NLRP3信号通路介导内皮细胞焦亡,进而改善系统性血管炎。展开更多
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 optio...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.展开更多
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...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.展开更多
Objective:To study the effects of Shenqi Tiaoshen Formula(SQTS)on the inflammatory response of MH-S cells induced by cigarette smoking extract(CSE)and its mechanism based on TLR4/NF-kB/NLRP3 pathway.Methods:MH-S cells...Objective:To study the effects of Shenqi Tiaoshen Formula(SQTS)on the inflammatory response of MH-S cells induced by cigarette smoking extract(CSE)and its mechanism based on TLR4/NF-kB/NLRP3 pathway.Methods:MH-S cells were used as subjects to evaluate cell viability by CCK-8 method.The levels of TNF-α,IL-1βand IL-6 in the supernatant were detected by ELISA.ROS were detected by DCFH-DA fluorescence probe.Western blotting was used to detect the expression of TLR4/NF-kB/NLRP3 pathway protein,and TAK-242,a TLR4 inhibitor,was used to verify the role of SQTS in the TLR4/NF-kB/NLRP3 pathway.Results:Compared with blank group,the cell survival rate of CSE group was decreased,and the contents of inflammatory cytokines TNF-α,IL-1βand IL-6 were increased(P<0.05),ROS fluorescence expression level was significantly increased(P<0.01),TLR4/NF-kB/NLRP3 pathway protein expression was significantly increased(P<0.05);Compared with CSE group,the survival rate of cells in SQTS groups was increased,and the expression levels of the above indexes were decreased(P<0.05),and TLR4/NF-kB/NLRP3 pathway protein decreased in TAK-242 groups(P<0.05).Conclusion:SQTS can reduce the inflammatory response of MH-S cells induced by CSE by inhibiting TLR4/NF-kB/NLRP3 pathway.展开更多
文摘目的研究二甲双胍(metformin,Met)通过抑制NF-κBNLRP3通路改善草酸钙(Calcium Oxalate,CaOx)诱导人肾小管上皮细胞(HK-2)损伤。方法分别随机将人肾小管上皮细胞分成对照组、实验组(CaOx+HK-2)、高浓度Met组(CaOx+HK-2+1.2 mM Met)、低浓度Met组(CaOx+HK-2+0.80 mM Met)、通路干预组(CaOx+HK-2+PDTC)五组。各组细胞培养24 h,采用酶联免疫吸附测定(Elisa)法检测细胞上清液中炎症因子(IL-6、IL-18、IL-1β),采用逆转录-聚合酶链反应(RT-PCR)法检测细胞中NF-κB、NLRP3、骨桥蛋白(OPN)mRNA。结果各组间细胞上清液IL-6、IL-18、IL-1β含量相比差异均有统计学意义(P均<0.05);各组间细胞中NF-κBmRNA表达量、NLRP3mRNA表达量、OPNmRNA表达量相比差异均有统计学意义(P均<0.05)。结论1.20 mmol/L或0.80 mmol/L Met可以通过抑制NF-κB/NLRP3通路调控下游的炎症相关蛋白及黏附蛋白表达,继而改善CaOx诱导人肾小管上皮细胞损伤及减少草酸钙肾结石形成。
基金reviewed and approved by the Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine Anhui Hospital Institutional Review Board(2022AH-022).
文摘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.
文摘目的探究高迁移率族蛋白1(High Mobility Group Box Protein1, HMGB1)通过(Toll-like Receptor4,TLR4)/核因子κB(Nuclear Factor Kappa-B, NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NOD-like Receptor Thermal Protein Domain Associated Protein 3, NLRP3)信号通路介导内皮细胞焦亡在系统性血管炎中的作用机制。方法 研究于2021年10月—2023年9月在齐齐哈尔医学院附属第三医院开展。取人脐静脉血管内皮细胞进行培养,随机分为对照组和实验组,实验组加入人重组HMGB1,对比对照组与实验组、实验组NLRP3及TLR4表达抑制前后,内皮细胞焦亡相关蛋白的表达水平。结果 系统性血管炎组与健康对照组比较,人脐静脉血管内皮细胞中HMGB1、含半胱氨酸的天冬氨酸蛋白水解酶-1(caspase-1)、白细胞介素-1β(interleukin-1β, IL-1β)、白细胞介素-18(interleukin-18, IL-18)水平升高,差异有统计学意义(P均<0.05)。细胞实验中实验组与对照组比较,caspase-1、IL-1β、IL-18水平升高,差异有统计学意义(P均<0.05)。过表达HMGB1并抑制NLRP3可使NLRP3(2.71±0.59 vs 1.24±0.58)、caspase-1(0.69±0.12 vs 0.40±0.03)、IL-1β[(1.75±0.31)pg/mL vs (1.16±0.12)pg/mL]、IL-18[(0.15±0.04)pg/mL vs (0.09±0.01)pg/mL]水平降低,差异有统计学意义(P均<0.05);过表达HMGB1并抑制TLR4可使TLR4(4.93±1.04 vs 1.96±0.84)、NF-κB(5.62±1.39 vs 2.15±1.04)、NLRP3(2.71±0.59 vs 1.24±0.58)、caspase-1(0.69±0.12 vs 0.40±0.03)、IL-1β[(1.75±0.31)pg/mL vs (1.16±0.12)pg/mL]、IL-18[(0.15±0.04)pg/mL vs (0.09±0.01)pg/mL]水平明显降低,差异有统计学意义(P均<0.05)。结论 HMGB1通过调节TLR4/NF-κB/NLRP3信号通路介导内皮细胞焦亡,进而改善系统性血管炎。
基金the National Key Research and Development Program of China,No.2017YFC0908104National Science and Technology Projects,No.2017ZX10203201,No.2017ZX10201201,and No.2017ZX10202202.
文摘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.
基金Heilongjiang Province Tradit Chin Med Research Projec(No.ZHY19-006)。
文摘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.
基金Regional Innovation Development Joint Fund of National Natural Science Foundation of China(No.U20A20398)Clinical Medical Research Transformation Project of Anhui Provincial Science and Technology Department(No.202204295107020045)。
文摘Objective:To study the effects of Shenqi Tiaoshen Formula(SQTS)on the inflammatory response of MH-S cells induced by cigarette smoking extract(CSE)and its mechanism based on TLR4/NF-kB/NLRP3 pathway.Methods:MH-S cells were used as subjects to evaluate cell viability by CCK-8 method.The levels of TNF-α,IL-1βand IL-6 in the supernatant were detected by ELISA.ROS were detected by DCFH-DA fluorescence probe.Western blotting was used to detect the expression of TLR4/NF-kB/NLRP3 pathway protein,and TAK-242,a TLR4 inhibitor,was used to verify the role of SQTS in the TLR4/NF-kB/NLRP3 pathway.Results:Compared with blank group,the cell survival rate of CSE group was decreased,and the contents of inflammatory cytokines TNF-α,IL-1βand IL-6 were increased(P<0.05),ROS fluorescence expression level was significantly increased(P<0.01),TLR4/NF-kB/NLRP3 pathway protein expression was significantly increased(P<0.05);Compared with CSE group,the survival rate of cells in SQTS groups was increased,and the expression levels of the above indexes were decreased(P<0.05),and TLR4/NF-kB/NLRP3 pathway protein decreased in TAK-242 groups(P<0.05).Conclusion:SQTS can reduce the inflammatory response of MH-S cells induced by CSE by inhibiting TLR4/NF-kB/NLRP3 pathway.