OBJECTIVE:To explore the mechanisms by which Huoxue Chubi decoction(活血除痹汤,HXCB) affects the protein kinase B(Akt)-mammalian target of rapamycin(mTOR) autophagy pathway in scleroderma Balb/c model mice.METHODS:A s...OBJECTIVE:To explore the mechanisms by which Huoxue Chubi decoction(活血除痹汤,HXCB) affects the protein kinase B(Akt)-mammalian target of rapamycin(mTOR) autophagy pathway in scleroderma Balb/c model mice.METHODS:A scleroderma model was established in male Balb/c mice,followed by daily administration of HXCB(4.6,2.3 and 1.15 g·kg^(-1)·d^(-1)) for 4 weeks.Bodyweight,epidermal and dermal thickness,dermal collagen levels,cutaneous reactive oxygen species(ROS) levels,Akt,Phosphorylated Akt(p-Akt),m TOR,Phosphorylated mTOR(p-mTOR),B-celllymphoma-2-interacting myosin-like coiled-coil protein 1(Beclin-1) and microtubule-associated protein A/B-light chain 3(LC3) protein and messenger ribonucleic acid(mRNA) expression were assessed.RESULTS:HXCB treatment significantly reduced epidermal and dermal thickness,dermal collagen levels,ROS levels and the mRNA and protein expression of factors in the Akt-mTOR signaling pathway compared to the scleroderma model group.Conversely,mice body weight and autophagy factors Beclin-1 and LC3 were significantly increased in mice receiving HXCB treatment.Moreover,finally,ROS expression positively correlated with skin thickness,collagen contents and the mRNA expression levels of Akt,while the protein and mRNA expression levels of Akt-mTOR pathway-related factors were inversely correlated with the protein and mRNA expression of Beclin-1 and LC3.CONCLUSION:HXCB can regulate autophagy by invigorating Qi and promoting blood circulation,thereby reducing blood stasis,facilitating new tissue generation,and contributing to scleroderma treatment.This effect may be attributed to the promotion of autophagy and enhancement of collagen degradation through the reduction of tissue oxidative stress elicited by HXCB.展开更多
Objective:To elucidate the effects of Huangqin Qingre Chubi capsule(HQC)on bone metabolism and serum TLR4 and NF-kB in rats with rheumatoid arthritis(RA),and to provide experimental bases for the treatment of RA with ...Objective:To elucidate the effects of Huangqin Qingre Chubi capsule(HQC)on bone metabolism and serum TLR4 and NF-kB in rats with rheumatoid arthritis(RA),and to provide experimental bases for the treatment of RA with HQC.Methods:A total of 40 SD rats were randomly divided into the normal group,model group,western medicine treatment group,and HQC group,with 10 rats in each group,and the RA model was induced by complete adjuvant in all groups except the normal group.After successful modeling,they were treated for 4 weeks to compare the degree of joint swelling;meanwhile,micro-CT was used to evaluate bone microstructural parameters,and changes in serum TLR4 and NF-kB expression levels were detected by ELISA.Results:There was no statistical significance in comparing the degree of joint swelling among the model group,western medicine treatment group and HQC group(P>0.05);Micro-CT results showed that bone microstructural parameters deteriorated in the osteoporosis model group compared with the normal group.Both western medicine and traditional Chinese medicine HQC could improve the bone microjunction of RA rats,and the HQC group was better than the western medicine treatment in improving the number of bone trabeculae(2.58±0.19)·mm^(-1);the serum results showed that RA was accompanied by the up-regulation of the expression of TLR4 and NF-kB.Both western drugs and HQC down-regulated the high expression of serum TLR4 and NF-kB in osteoporotic rats,and the down-regulation of serum TLR4(9.90±0.55)ng·ml^(-1)and NF-kB(350.29±3.14)ng·L^(-1)by HQC was more obvious.Conclusion:Chinese herbal medicine HQC can significantly improve the bone microstructure of RA rats,and its effect is better than that of western medicine in some aspects.The mechanism of action of HQC is related to the inhibition of the activation of the TLR4/NF-kB pathway,and this study provides an experimental basis for the further development and utilization of HQC.展开更多
基金Natural Science Foundation-funded Project:Exploration the Mechanism of Yiqi Huoxue Therapy in Treating Scleroderma Fibrosis based on Phosphatidylinositol 3-Kinase (PI3K)-Protein Kinase B (Akt)-Mammalian Target of Rapamycin (mTOR) Signal Pathway about Autophagy (No.81804106)the 2023 Science and Technology Innovation Project Dongzhimen Hospital Beijing University of Chinese Medicine:Exploration the Mechanism of Radix Salviae Miltiorrhizae Components in Treating Scleroderma Fibrosis Based on PI3K-Akt-mTOR Signal Pathway about Autophagy (No.DZMKJCX-2023-009)。
文摘OBJECTIVE:To explore the mechanisms by which Huoxue Chubi decoction(活血除痹汤,HXCB) affects the protein kinase B(Akt)-mammalian target of rapamycin(mTOR) autophagy pathway in scleroderma Balb/c model mice.METHODS:A scleroderma model was established in male Balb/c mice,followed by daily administration of HXCB(4.6,2.3 and 1.15 g·kg^(-1)·d^(-1)) for 4 weeks.Bodyweight,epidermal and dermal thickness,dermal collagen levels,cutaneous reactive oxygen species(ROS) levels,Akt,Phosphorylated Akt(p-Akt),m TOR,Phosphorylated mTOR(p-mTOR),B-celllymphoma-2-interacting myosin-like coiled-coil protein 1(Beclin-1) and microtubule-associated protein A/B-light chain 3(LC3) protein and messenger ribonucleic acid(mRNA) expression were assessed.RESULTS:HXCB treatment significantly reduced epidermal and dermal thickness,dermal collagen levels,ROS levels and the mRNA and protein expression of factors in the Akt-mTOR signaling pathway compared to the scleroderma model group.Conversely,mice body weight and autophagy factors Beclin-1 and LC3 were significantly increased in mice receiving HXCB treatment.Moreover,finally,ROS expression positively correlated with skin thickness,collagen contents and the mRNA expression levels of Akt,while the protein and mRNA expression levels of Akt-mTOR pathway-related factors were inversely correlated with the protein and mRNA expression of Beclin-1 and LC3.CONCLUSION:HXCB can regulate autophagy by invigorating Qi and promoting blood circulation,thereby reducing blood stasis,facilitating new tissue generation,and contributing to scleroderma treatment.This effect may be attributed to the promotion of autophagy and enhancement of collagen degradation through the reduction of tissue oxidative stress elicited by HXCB.
文摘基于超高效液相色谱-四极杆静电场轨道阱质谱(UPLC-Q-Exactive Orbitrap-MS)技术及网络药理学探讨化浊散结除痹方治疗痛风性关节炎(gouty arthritis,GA)的药效物质及潜在机制。采用UPLC-Q-Exactive Orbitrap-MS技术鉴定化浊散结除痹方药物成分,对其有效成分进行定性分析,共鉴定出化浊散结除痹方中184个有效成分;通过PharmMapper在线数据库筛选有效成分靶点897个,在OMIM、GeneCards、CTD等数据库获取GA相关的疾病靶点491个,进行韦恩分析后获得二者的交集靶点60个,通过Cytoscape平台构建“成分靶点-GA靶点”网络图,利用STRING数据库构建蛋白-蛋白互作网络,筛选出16个核心靶点,将核心靶点进行基因本体论(Gene Ontology,GO)与京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路的富集分析,并构建“成分-靶点-通路”网络图,发现该方治疗GA的主要有效成分为酚类、黄酮类、生物碱类、萜类化合物,关键靶点有SRC、MMP3、MMP9、REN、ALB、IGF1R、PPARG、MAPK1、HPRT1、CASP1,通过GO分析发现其治疗GA主要涉及脂质反应、细菌反应、生物刺激的反应等生物过程,通过KEGG分析发现其治疗GA相关的通路有脂质和动脉粥样硬化、中性粒细胞胞外诱捕网、IL-17等。综上,该研究揭示了酚类、黄酮类、生物碱类、萜类化合物可能是化浊散结除痹方治疗GA的核心药效物质,其药效机制可能与SRC、MMP3、MMP9等靶点及脂质和动脉粥样硬化、中性粒细胞胞外诱捕网、IL-17等通路相关。
文摘Objective:To elucidate the effects of Huangqin Qingre Chubi capsule(HQC)on bone metabolism and serum TLR4 and NF-kB in rats with rheumatoid arthritis(RA),and to provide experimental bases for the treatment of RA with HQC.Methods:A total of 40 SD rats were randomly divided into the normal group,model group,western medicine treatment group,and HQC group,with 10 rats in each group,and the RA model was induced by complete adjuvant in all groups except the normal group.After successful modeling,they were treated for 4 weeks to compare the degree of joint swelling;meanwhile,micro-CT was used to evaluate bone microstructural parameters,and changes in serum TLR4 and NF-kB expression levels were detected by ELISA.Results:There was no statistical significance in comparing the degree of joint swelling among the model group,western medicine treatment group and HQC group(P>0.05);Micro-CT results showed that bone microstructural parameters deteriorated in the osteoporosis model group compared with the normal group.Both western medicine and traditional Chinese medicine HQC could improve the bone microjunction of RA rats,and the HQC group was better than the western medicine treatment in improving the number of bone trabeculae(2.58±0.19)·mm^(-1);the serum results showed that RA was accompanied by the up-regulation of the expression of TLR4 and NF-kB.Both western drugs and HQC down-regulated the high expression of serum TLR4 and NF-kB in osteoporotic rats,and the down-regulation of serum TLR4(9.90±0.55)ng·ml^(-1)and NF-kB(350.29±3.14)ng·L^(-1)by HQC was more obvious.Conclusion:Chinese herbal medicine HQC can significantly improve the bone microstructure of RA rats,and its effect is better than that of western medicine in some aspects.The mechanism of action of HQC is related to the inhibition of the activation of the TLR4/NF-kB pathway,and this study provides an experimental basis for the further development and utilization of HQC.
