Summary:Huai Qi Huang(HQH)exerts great effects in clinic,such as anti-inflammation,immune-regulation,anti-cancer,and so on.However,the mechanism by which HQH protects juvenile idiopathic arthritis(JIA)is obscure.Thus,...Summary:Huai Qi Huang(HQH)exerts great effects in clinic,such as anti-inflammation,immune-regulation,anti-cancer,and so on.However,the mechanism by which HQH protects juvenile idiopathic arthritis(JIA)is obscure.Thus,we explored deeply the protective mechanisms in juvenile collagen-induced arthritis(CIA)rat model.Pyroptosis is Gasdermin D(GSDMD)-dependent programmed cell death,involved in many diseases,such as sepsis.We investigated whether GSDMD-induced pyroptosis take part in mechanisms of juvenile CIA arthritis.Juvenile Wistar rats(3-4 weeks)were injected intradermally with fully emulsified bovine typeⅡcollagen and complete Freund's adjuvant to establish CIA rat models.Later,the CIA rats received oral administration of HQH(4.16 g/kg)once a day from the day 21 of modeling,with the treatment lasting for 28 days.Varieties of indicators were measured for evaluation of anti-inflammation effect of HQH,including hind paw swelling,arthritis scores,micro CT,and histopathological changes and the level of pro-inflammatory cytokines in the serum,including tumor necrosis factor alpha(TNF-α)and interleukin-18(IL-18).The expression of GSDMD and caspasein the joint synovial tissues was detected.The results demonstrated that the expression of the pyroptotic protein GSDMD and its upstream caspase-1 was significantly increased in the synovial tissues of CIA rats.The treatment of HQH ameliorated the symptoms in CIA rats,reduced levels of pro-inflammatory cytokines and hind paw swelling,down-regulated the expression of GDSMD and caspase-1.GSDMDinduced pyroptosis participated in the pathogenesis of CIA rats.The study supported that HQH can effectively improve joints inflammation of juvenile collagen-induced arthritis rats by inhibiting pyroptosis pathway in the joint synovial tissues.展开更多
Imatinib mesylate(IM)is the first-line treatment for Philadelphia(Ph)chromosomal positive leukemia by inhibiting phosphorylation of substrates via binding to the ABL kinase domain.Because of the drug resistance,side e...Imatinib mesylate(IM)is the first-line treatment for Philadelphia(Ph)chromosomal positive leukemia by inhibiting phosphorylation of substrates via binding to the ABL kinase domain.Because of the drug resistance,side effects and the high cost of IM,it is necessary to find anti-cancer drugs with relatively low toxicity and cost,and enhanced efficacy,such as traditional Chinese medicines(TCMs).As one of TCMs,Huai Qi Huang(HQH)was chosen to treat BV173 and K562 cells.Various concentrations of HQH were added to cells for 24-72 h.Co-treatment of HQH and trametinib,an MEK inhibitor,was used to verify the synergistic effects on cell viability and apoptosis.Knockdown and overexpression of mitogen-activated protein kinase kinase 4(MEK4)were implemented to demonstrate the role of MEK in cell apoptosis.Cell viability and apoptosis were measured by cell counting kit-8 assay(CCK8)and flow cytometry,respectively.Western blotting and real-time quantitative PCR(RT-qPCR)were used to assess protein and mRNA expression levels,respectively.The results showed that HQH inhibited survival and promoted apoptosis of BV173 and K562 cells in a dose-dependent manner,accompanied with down-regulation of PRKCH mRNA as well as CRAF,MEK4,phospho-ERK(pERK)and BCL2 proteins,and up-regulation of cleaved caspase3 protein.Co-treatment of HQH and trametinib had.a synergistic effect on inhibiting survival and promoting apoptosis.MEK4 knockdown increased apoptosis,and had a synergistic effeet with HQH.In contrast,MEK4 overexpression decreased.apoptosis,and had the opposite efect with HQH.Collectively,the results of this study may identify a therapeutic mechanism of HQH on promoting apoptosis,and provide a potential option for treatment of Pht leukemia.展开更多
基金This study was supported by the National Natural Science Foundation of China(No.81270949).
文摘Summary:Huai Qi Huang(HQH)exerts great effects in clinic,such as anti-inflammation,immune-regulation,anti-cancer,and so on.However,the mechanism by which HQH protects juvenile idiopathic arthritis(JIA)is obscure.Thus,we explored deeply the protective mechanisms in juvenile collagen-induced arthritis(CIA)rat model.Pyroptosis is Gasdermin D(GSDMD)-dependent programmed cell death,involved in many diseases,such as sepsis.We investigated whether GSDMD-induced pyroptosis take part in mechanisms of juvenile CIA arthritis.Juvenile Wistar rats(3-4 weeks)were injected intradermally with fully emulsified bovine typeⅡcollagen and complete Freund's adjuvant to establish CIA rat models.Later,the CIA rats received oral administration of HQH(4.16 g/kg)once a day from the day 21 of modeling,with the treatment lasting for 28 days.Varieties of indicators were measured for evaluation of anti-inflammation effect of HQH,including hind paw swelling,arthritis scores,micro CT,and histopathological changes and the level of pro-inflammatory cytokines in the serum,including tumor necrosis factor alpha(TNF-α)and interleukin-18(IL-18).The expression of GSDMD and caspasein the joint synovial tissues was detected.The results demonstrated that the expression of the pyroptotic protein GSDMD and its upstream caspase-1 was significantly increased in the synovial tissues of CIA rats.The treatment of HQH ameliorated the symptoms in CIA rats,reduced levels of pro-inflammatory cytokines and hind paw swelling,down-regulated the expression of GDSMD and caspase-1.GSDMDinduced pyroptosis participated in the pathogenesis of CIA rats.The study supported that HQH can effectively improve joints inflammation of juvenile collagen-induced arthritis rats by inhibiting pyroptosis pathway in the joint synovial tissues.
基金The present study was supported by the National Natural Science Foundation of China(No.81700147).
文摘Imatinib mesylate(IM)is the first-line treatment for Philadelphia(Ph)chromosomal positive leukemia by inhibiting phosphorylation of substrates via binding to the ABL kinase domain.Because of the drug resistance,side effects and the high cost of IM,it is necessary to find anti-cancer drugs with relatively low toxicity and cost,and enhanced efficacy,such as traditional Chinese medicines(TCMs).As one of TCMs,Huai Qi Huang(HQH)was chosen to treat BV173 and K562 cells.Various concentrations of HQH were added to cells for 24-72 h.Co-treatment of HQH and trametinib,an MEK inhibitor,was used to verify the synergistic effects on cell viability and apoptosis.Knockdown and overexpression of mitogen-activated protein kinase kinase 4(MEK4)were implemented to demonstrate the role of MEK in cell apoptosis.Cell viability and apoptosis were measured by cell counting kit-8 assay(CCK8)and flow cytometry,respectively.Western blotting and real-time quantitative PCR(RT-qPCR)were used to assess protein and mRNA expression levels,respectively.The results showed that HQH inhibited survival and promoted apoptosis of BV173 and K562 cells in a dose-dependent manner,accompanied with down-regulation of PRKCH mRNA as well as CRAF,MEK4,phospho-ERK(pERK)and BCL2 proteins,and up-regulation of cleaved caspase3 protein.Co-treatment of HQH and trametinib had.a synergistic effect on inhibiting survival and promoting apoptosis.MEK4 knockdown increased apoptosis,and had a synergistic effeet with HQH.In contrast,MEK4 overexpression decreased.apoptosis,and had the opposite efect with HQH.Collectively,the results of this study may identify a therapeutic mechanism of HQH on promoting apoptosis,and provide a potential option for treatment of Pht leukemia.
