Objectives:This study is to investigate the effects of new anti-tumor formular(NAF)on expression of PCNA,P21 ras and NF-KB P65 in liver precancerous lesions of HBV large envelope transgenic mice injected by aflatoxin ...Objectives:This study is to investigate the effects of new anti-tumor formular(NAF)on expression of PCNA,P21 ras and NF-KB P65 in liver precancerous lesions of HBV large envelope transgenic mice injected by aflatoxin B1(AFB1).Methods:The precancerous HBV large envelope transgenic mouse injected by AFB1 liver model was used.Mice was given water,NAF concentrated water solution throughout the whole experiment(48 weeks).PCNA,P21 ras and NF-KB P65 protein expression were detected by immunohistochemical method.Results: PCNA,P21 ras and NF-KB P65 expressions were significantly inhibited by NAF treatment.Conclusion NAF inhibited PCNA,P21 ras and NF-KB P65 protein expressions,therefore,NAF could show obvious effects on protecting against synergistic hepatoearcinogenesis of HBV and AFB1.展开更多
目的探讨沉默信息调节因子1(SIRT1)在异烟肼致人肝细胞损伤中的作用。方法培养人正常肝细胞HL-7702,实验分为6组:空白对照组、异烟肼组、异烟肼+SIRT1激动剂组、SIRT1激动剂对照组、异烟肼+SIRT1抑制剂组、SIRT1抑制剂对照组。取各组细...目的探讨沉默信息调节因子1(SIRT1)在异烟肼致人肝细胞损伤中的作用。方法培养人正常肝细胞HL-7702,实验分为6组:空白对照组、异烟肼组、异烟肼+SIRT1激动剂组、SIRT1激动剂对照组、异烟肼+SIRT1抑制剂组、SIRT1抑制剂对照组。取各组细胞上清液测定丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)含量;实时荧光定量聚合酶链反应(q RT-PCR)检测肝细胞SIRT1、NF-k B p65 mRNA表达水平;酶联免疫吸附法(ELISA)检测SIRT1、核转录因子k B(NF-k B p65)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)蛋白表达水平。结果与空白对照组比较,异烟肼组细胞SIRT1的mRNA和蛋白表达下降(P<0.05),其下游靶基因NF-k B p65的mRNA和蛋白表达升高(P<0.05),炎症因子IL-6、TNF-α蛋白表达水平升高(P<0.05)。加入SIRT1激动剂可减轻异烟肼引起的炎症反应,加入SIRT1抑制剂可使NF-k B p65、IL-6、TNF-α表达水平进一步升高从而加重细胞的炎症损伤。结论异烟肼诱导肝细胞损伤过程中,降低SIRT1水平,增加炎症因子的表达。SIRT1的激活可以通过降低NF-k B p65表达进而减轻肝细胞损伤的发生。展开更多
BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene ma...BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.展开更多
目的:探究清肺通络膏对流感病毒诱导的肺炎大鼠肺组织中PI3K/AKT/NF-KB信号传导通路的调控机制。方法:将40只Wistar大鼠随机分为正常组,模型组,清肺通络膏高、中、低剂量组,除正常组外,采用鼻腔接种流感病毒FM1株诱导Wistar幼龄大鼠肺炎...目的:探究清肺通络膏对流感病毒诱导的肺炎大鼠肺组织中PI3K/AKT/NF-KB信号传导通路的调控机制。方法:将40只Wistar大鼠随机分为正常组,模型组,清肺通络膏高、中、低剂量组,除正常组外,采用鼻腔接种流感病毒FM1株诱导Wistar幼龄大鼠肺炎,在感染后各治疗组采用清肺通络膏贴敷于大鼠背部肺脏投影区治疗。观察各组大鼠的一般状态;肺组织形态学;采用免疫组化染色法检测各组大鼠肺组织PI3K,AKT,核因子NFB p65的表达水平。结果:与正常组相比,模型组大鼠肺组织中PI3K,AKT,NF-k B p65的表达显著增高(P<0.05),清肺通络膏高剂量组PI3K,AKT,NF-k B p65的表达较模型组明显降低(P<0.05)。结论:清肺通络膏通过抑制PI3K/AKT信号通路,使NF-k B p65表达降低,从而减轻了肺组织的病理损伤,达到治疗目的。展开更多
文摘Objectives:This study is to investigate the effects of new anti-tumor formular(NAF)on expression of PCNA,P21 ras and NF-KB P65 in liver precancerous lesions of HBV large envelope transgenic mice injected by aflatoxin B1(AFB1).Methods:The precancerous HBV large envelope transgenic mouse injected by AFB1 liver model was used.Mice was given water,NAF concentrated water solution throughout the whole experiment(48 weeks).PCNA,P21 ras and NF-KB P65 protein expression were detected by immunohistochemical method.Results: PCNA,P21 ras and NF-KB P65 expressions were significantly inhibited by NAF treatment.Conclusion NAF inhibited PCNA,P21 ras and NF-KB P65 protein expressions,therefore,NAF could show obvious effects on protecting against synergistic hepatoearcinogenesis of HBV and AFB1.
文摘目的探讨沉默信息调节因子1(SIRT1)在异烟肼致人肝细胞损伤中的作用。方法培养人正常肝细胞HL-7702,实验分为6组:空白对照组、异烟肼组、异烟肼+SIRT1激动剂组、SIRT1激动剂对照组、异烟肼+SIRT1抑制剂组、SIRT1抑制剂对照组。取各组细胞上清液测定丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)含量;实时荧光定量聚合酶链反应(q RT-PCR)检测肝细胞SIRT1、NF-k B p65 mRNA表达水平;酶联免疫吸附法(ELISA)检测SIRT1、核转录因子k B(NF-k B p65)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)蛋白表达水平。结果与空白对照组比较,异烟肼组细胞SIRT1的mRNA和蛋白表达下降(P<0.05),其下游靶基因NF-k B p65的mRNA和蛋白表达升高(P<0.05),炎症因子IL-6、TNF-α蛋白表达水平升高(P<0.05)。加入SIRT1激动剂可减轻异烟肼引起的炎症反应,加入SIRT1抑制剂可使NF-k B p65、IL-6、TNF-α表达水平进一步升高从而加重细胞的炎症损伤。结论异烟肼诱导肝细胞损伤过程中,降低SIRT1水平,增加炎症因子的表达。SIRT1的激活可以通过降低NF-k B p65表达进而减轻肝细胞损伤的发生。
文摘BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.
文摘目的:探究清肺通络膏对流感病毒诱导的肺炎大鼠肺组织中PI3K/AKT/NF-KB信号传导通路的调控机制。方法:将40只Wistar大鼠随机分为正常组,模型组,清肺通络膏高、中、低剂量组,除正常组外,采用鼻腔接种流感病毒FM1株诱导Wistar幼龄大鼠肺炎,在感染后各治疗组采用清肺通络膏贴敷于大鼠背部肺脏投影区治疗。观察各组大鼠的一般状态;肺组织形态学;采用免疫组化染色法检测各组大鼠肺组织PI3K,AKT,核因子NFB p65的表达水平。结果:与正常组相比,模型组大鼠肺组织中PI3K,AKT,NF-k B p65的表达显著增高(P<0.05),清肺通络膏高剂量组PI3K,AKT,NF-k B p65的表达较模型组明显降低(P<0.05)。结论:清肺通络膏通过抑制PI3K/AKT信号通路,使NF-k B p65表达降低,从而减轻了肺组织的病理损伤,达到治疗目的。
基金supported by grants from The National Natural Science Foundation of China(81541163)The Natural Science Youth Foundation of Hunan Province(2015JJ3101)+1 种基金Training project of Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study(2014-405)The Construct Program of The Key Discipline in Hunan Province~~
