目的探讨柴朴汤对哮喘小鼠支气管上皮细胞高迁移率族蛋白B1(HMGB1)、Toll样受体4(TLR4)、核因子-κB(NF-κB)及下游炎症介质单核细胞趋化蛋白-1(MCP-1)、白细胞介素-6(IL-6)的影响。方法将小鼠支气管上皮细胞分为对照组(Con组,空白血清...目的探讨柴朴汤对哮喘小鼠支气管上皮细胞高迁移率族蛋白B1(HMGB1)、Toll样受体4(TLR4)、核因子-κB(NF-κB)及下游炎症介质单核细胞趋化蛋白-1(MCP-1)、白细胞介素-6(IL-6)的影响。方法将小鼠支气管上皮细胞分为对照组(Con组,空白血清培养)、Model组(哮喘小鼠血清培养)和Chaipu组(柴朴汤药血清培养)。采用qRT-PCR和Western blotting检测小鼠支气管上皮细胞中HMGB1、TLR4、NF-κB、MCP-1、IL-6的mRNA及蛋白表达情况,ELISA法测定细胞培养液中MCP-1和IL-6的水平。结果与Con组相比,Model组小鼠支气管上皮细胞HMGB1、TLR4、NF-κB、MCP-1、IL-6表达均明显升高(P<0.05);Model组细胞培养液中MCP-1、IL-6也显著升高(P<0.05)。与Model组相比,Chaipu组小鼠支气管上皮细胞HMGB1、TLR4、MCP-1、IL-6 mRNA和蛋白表达降低,NF-κB m RNA表达亦降低(P<0.05);Chaipu组细胞培养液中MCP-1、IL-6也显著降低(P<0.05)。结论柴朴汤可下调哮喘血清诱导的炎症因子的表达,可能与其抑制HMGB1/TLR4/NF-κB炎症通路有关。展开更多
【目的】研究苦苣菜水提物对脂多糖(LPS)诱导的小鼠肺炎模型中炎症相关因子及炎症信号通路的影响,探究苦苣菜水提物对小鼠肺炎的抗炎作用。【方法】制备苦苣菜水提物,将昆明小鼠随机分为空白组、模型组、阳性对照组及苦苣菜水提物低、...【目的】研究苦苣菜水提物对脂多糖(LPS)诱导的小鼠肺炎模型中炎症相关因子及炎症信号通路的影响,探究苦苣菜水提物对小鼠肺炎的抗炎作用。【方法】制备苦苣菜水提物,将昆明小鼠随机分为空白组、模型组、阳性对照组及苦苣菜水提物低、中、高浓度组,空白组小鼠不做任何处理,模型组小鼠灌胃0.3 mL生理盐水,阳性对照组小鼠灌胃0.3 mL 5 mg/kg地塞米松,苦苣菜水提物低、中、高浓度组小鼠依次灌胃0.3 mL 100、200和400 mg/mL苦苣菜水提物,持续灌胃给药1周后,空白组小鼠腹腔注射0.3 mL生理盐水,其余组小鼠腹腔注射0.3 mL 30 mg/kg LPS诱导建立小鼠肺炎模型。采用ELISA方法检测各组小鼠血清中白细胞介素-6(IL-6)、IL-10、转化生长因子-β(TGF-β)的含量;利用免疫组织化学(IHC)和Western blotting检测各组小鼠肺中炎症信号通路(HMGB1/TLR4/NF-κB)蛋白的表达水平。【结果】与空白组相比,模型组小鼠血清中IL-6含量显著上调(P<0.05),IL-10、TGF-β含量有所上调,肺脏中HMGB1、TLR4、NF-κB的分布及蛋白表达量极显著上调(P<0.01);与模型组相比,苦苣菜水提物处理组小鼠血清中促炎因子IL-6含量有所下调,抗炎因子IL-10含量有所上调,抗炎因子TGF-β含量极显著上调(P<0.01);在肺脏中HMGB1、TLR4、NF-κB大量分布在肺泡周围的上皮细胞上,与模型组相比,苦苣菜水提物处理组小鼠肺脏中HMGB1、TLR4、NF-κB的分布极显著下调(P<0.01),且表现出剂量依赖效应,HMGB1、TLR4蛋白表达量极显著下调(P<0.01),NF-κB蛋白表达量显著或极显著下调(P<0.05;P<0.01)。【结论】苦苣菜水提物能抑制LPS诱导的肺炎小鼠血清中促炎因子IL-6的分泌及肺脏中信号通路HMGB1、TLR4、NF-κB蛋白的表达,促进小鼠血清中抗炎因子IL-10、TGF-β的分泌,表明苦苣菜水提物具有抗肺炎作用。展开更多
Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic ne...Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells was detected by RT-qPCR.Then,the TRPM7 interference vector was constructed,and the downstream high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway proteins were detected.Next,in addition to interference with TRPM7 expression,overexpression of HMGB1 in high glucose-induced HK-2 cells was performed.Cell activity,apoptosis,oxidative stress levels,and inflammation levels were determined by CCK8,TUNEL,Western blotting,immunofluorescence and related kits.Results:TRPM7 expression was upregulated in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells.Interference with TRPM7 reduced cell damage,epithelial-mesenchymal transition,oxidative stress,and inflammatory response in high glucose-induced HK-2 cells via inhibiting the HMGB1/TLR4 signaling pathway.However,the effects induced by TRPM7 silencing were abrogated by HMGB1 overexpression.Conclusions:Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway.Further animal experiments and clinical trials are warranted to verify its effect.展开更多
文摘目的探讨柴朴汤对哮喘小鼠支气管上皮细胞高迁移率族蛋白B1(HMGB1)、Toll样受体4(TLR4)、核因子-κB(NF-κB)及下游炎症介质单核细胞趋化蛋白-1(MCP-1)、白细胞介素-6(IL-6)的影响。方法将小鼠支气管上皮细胞分为对照组(Con组,空白血清培养)、Model组(哮喘小鼠血清培养)和Chaipu组(柴朴汤药血清培养)。采用qRT-PCR和Western blotting检测小鼠支气管上皮细胞中HMGB1、TLR4、NF-κB、MCP-1、IL-6的mRNA及蛋白表达情况,ELISA法测定细胞培养液中MCP-1和IL-6的水平。结果与Con组相比,Model组小鼠支气管上皮细胞HMGB1、TLR4、NF-κB、MCP-1、IL-6表达均明显升高(P<0.05);Model组细胞培养液中MCP-1、IL-6也显著升高(P<0.05)。与Model组相比,Chaipu组小鼠支气管上皮细胞HMGB1、TLR4、MCP-1、IL-6 mRNA和蛋白表达降低,NF-κB m RNA表达亦降低(P<0.05);Chaipu组细胞培养液中MCP-1、IL-6也显著降低(P<0.05)。结论柴朴汤可下调哮喘血清诱导的炎症因子的表达,可能与其抑制HMGB1/TLR4/NF-κB炎症通路有关。
文摘【目的】研究苦苣菜水提物对脂多糖(LPS)诱导的小鼠肺炎模型中炎症相关因子及炎症信号通路的影响,探究苦苣菜水提物对小鼠肺炎的抗炎作用。【方法】制备苦苣菜水提物,将昆明小鼠随机分为空白组、模型组、阳性对照组及苦苣菜水提物低、中、高浓度组,空白组小鼠不做任何处理,模型组小鼠灌胃0.3 mL生理盐水,阳性对照组小鼠灌胃0.3 mL 5 mg/kg地塞米松,苦苣菜水提物低、中、高浓度组小鼠依次灌胃0.3 mL 100、200和400 mg/mL苦苣菜水提物,持续灌胃给药1周后,空白组小鼠腹腔注射0.3 mL生理盐水,其余组小鼠腹腔注射0.3 mL 30 mg/kg LPS诱导建立小鼠肺炎模型。采用ELISA方法检测各组小鼠血清中白细胞介素-6(IL-6)、IL-10、转化生长因子-β(TGF-β)的含量;利用免疫组织化学(IHC)和Western blotting检测各组小鼠肺中炎症信号通路(HMGB1/TLR4/NF-κB)蛋白的表达水平。【结果】与空白组相比,模型组小鼠血清中IL-6含量显著上调(P<0.05),IL-10、TGF-β含量有所上调,肺脏中HMGB1、TLR4、NF-κB的分布及蛋白表达量极显著上调(P<0.01);与模型组相比,苦苣菜水提物处理组小鼠血清中促炎因子IL-6含量有所下调,抗炎因子IL-10含量有所上调,抗炎因子TGF-β含量极显著上调(P<0.01);在肺脏中HMGB1、TLR4、NF-κB大量分布在肺泡周围的上皮细胞上,与模型组相比,苦苣菜水提物处理组小鼠肺脏中HMGB1、TLR4、NF-κB的分布极显著下调(P<0.01),且表现出剂量依赖效应,HMGB1、TLR4蛋白表达量极显著下调(P<0.01),NF-κB蛋白表达量显著或极显著下调(P<0.05;P<0.01)。【结论】苦苣菜水提物能抑制LPS诱导的肺炎小鼠血清中促炎因子IL-6的分泌及肺脏中信号通路HMGB1、TLR4、NF-κB蛋白的表达,促进小鼠血清中抗炎因子IL-10、TGF-β的分泌,表明苦苣菜水提物具有抗肺炎作用。
文摘Objective:To explore the regulatory mechanism of transient receptor potential melastatin-7(TRPM7)in high glucose-induced renal tubular epithelial cell injury.Methods:The expression of TRPM7 in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells was detected by RT-qPCR.Then,the TRPM7 interference vector was constructed,and the downstream high mobility group box 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway proteins were detected.Next,in addition to interference with TRPM7 expression,overexpression of HMGB1 in high glucose-induced HK-2 cells was performed.Cell activity,apoptosis,oxidative stress levels,and inflammation levels were determined by CCK8,TUNEL,Western blotting,immunofluorescence and related kits.Results:TRPM7 expression was upregulated in the serum of diabetic nephropathy patients and high glucose-induced HK-2 cells.Interference with TRPM7 reduced cell damage,epithelial-mesenchymal transition,oxidative stress,and inflammatory response in high glucose-induced HK-2 cells via inhibiting the HMGB1/TLR4 signaling pathway.However,the effects induced by TRPM7 silencing were abrogated by HMGB1 overexpression.Conclusions:Decreased TRPM7 alleviates high glucose-induced renal tubular epithelial cell injury by inhibiting the HMGB1/TLR4 signaling pathway.Further animal experiments and clinical trials are warranted to verify its effect.
