Catalpol is the main active ingredient of an extract from Radix rehmanniae,which in a previous study showed a protective effect against various types of tissue injury.However,a protective effect of catalpol on uterine...Catalpol is the main active ingredient of an extract from Radix rehmanniae,which in a previous study showed a protective effect against various types of tissue injury.However,a protective effect of catalpol on uterine inflammation has not been reported.In this study,to investigate the protective mechanism of catalpol on lipopolysaccharide(LPS)-induced bovine endometrial epithelial cells(bEECs)and mouse endometritis,in vitro and in vivo inflammation models were established.The Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway and its downstream inflammatory factors were detected by enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qRT-PCR),western blot(WB),and immunofluorescence techniques.The results from ELISA and qRT-PCR showed that catalpol dose-dependently reduced the expression of pro-inflammatory cytokines such as tumor necrosis factorα(TNF-α),interleukin(IL)-1β,and IL-6,and chemokines such as C-X-C motif chemokine ligand 8(CXCL8)and CXCL5,both in bEECs and in uterine tissue.From the experimental results of WB,qRT-PCR,and immunofluorescence,the expression of TLR4 and the phosphorylation of NF-κB p65 were markedly inhibited by catalpol compared with the LPS group.The inflammatory damage to the mouse uterus caused by LPS was greatly reduced and was accompanied by a decline in myeloperoxidase(MPO)activity.The results of this study suggest that catalpol can exert an anti-inflammatory impact on LPS-induced bEECs and mouse endometritis by inhibiting inflammation and activation of the TLR4/NF-κB signaling pathway.展开更多
Ds-echinoside A (DSEA),a non-sulfated triterpene glycoside,was isolated from the sea cucumber Pearsonothuria graeffei.In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion,mi...Ds-echinoside A (DSEA),a non-sulfated triterpene glycoside,was isolated from the sea cucumber Pearsonothuria graeffei.In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion,migration,invasion,and angiogenesis.In this study,we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2,with a half-maximal inhibitory concentration (IC50) of 2.65 μmol/L,and suppressed Hep G2 cell adhesion,migration,and invasion in a dose-dependent manner.DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane (CAM) assay in vivo.Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 (MMP-9),which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis.DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1),an important regulator of MMP-9 activation.From the results of Western blotting,the expressions of nuclear factor-kappa B (NF-κB) and vascular endothelial growth factor (VEGF) were found to be remarkably reduced by DSEA.These findings suggest that DSEA exhibits a significant antimetastatic activity through the specific inhibition of NF-κB-dependent MMP-9 and VEGF expressions.展开更多
目的:观察柴胡疏肝散对非酒精性脂肪性肝炎(NASH)大鼠肝脏内质网应激(ERS)相关IRE-1α/NF-κB通路的影响,探讨其防治NASH的可能作用机制。方法:采用高脂饮食12周建立NASH大鼠模型,同时以6g·kg-1·d-1的柴胡疏肝散干预12周,油红...目的:观察柴胡疏肝散对非酒精性脂肪性肝炎(NASH)大鼠肝脏内质网应激(ERS)相关IRE-1α/NF-κB通路的影响,探讨其防治NASH的可能作用机制。方法:采用高脂饮食12周建立NASH大鼠模型,同时以6g·kg-1·d-1的柴胡疏肝散干预12周,油红O和HE染色光镜观察肝组织脂肪变和炎性反应程度;计算NAFLD活动度(NAS);生化法检测血清ALT、AST和肝组织TG、GHOL;Realtime-PCR和Western blot技术检测肝组织GRP78、IRE-1α、NF-κB m RNA和蛋白或磷酸化水平,ELISA检测血清和肝组织中TNF-α、IL-1β水平。结果:模型组和柴胡疏肝散组大鼠肝组织均出现了脂肪变和炎性坏死。与正常组比较,模型组大鼠NAS显著增高(P<0.01),血清ALT、AST水平和肝组织TG、CHOL含量显著升高(P<0.01),肝组织GRP78、IRE-1α、NF-κB m RNA表达显著增强(P<0.01),GRP78蛋白表达和IRE-1α、NF-κB m RNA磷酸化水平显著上调(P<0.01),血清及肝组织TNF-α、IL-1β含量显著增加(P<0.01)。与模型组比较,柴胡疏肝散组大鼠NAS显著降低(P<0.01),血清ALT、AST水平和肝组织TG、CHOL含量显著降低(P<0.01),肝组织GRP78、IRE-1α、NF-κB m RNA表达显著下降(P<0.05,P<0.01),GRP78蛋白表达和IRE-1α、NF-κB m RNA磷酸化水平显著下调(P<0.05),血清及肝组织TNF-α、IL-1β含量显著下降(P<0.05)。结论:ERS相关的IRE-1α/NF-κB参与高脂饮食诱导的大鼠NASH的发生发展。柴胡疏肝散可能通过调节IRE-1α/NF-κB通路抑制过度的ERS,来调节肝脏脂质代谢紊乱、减轻肝脏炎性反应,防止NASH的进展。展开更多
基金Project supported by the National Natural Science Foundation of China(No.31472254)
文摘Catalpol is the main active ingredient of an extract from Radix rehmanniae,which in a previous study showed a protective effect against various types of tissue injury.However,a protective effect of catalpol on uterine inflammation has not been reported.In this study,to investigate the protective mechanism of catalpol on lipopolysaccharide(LPS)-induced bovine endometrial epithelial cells(bEECs)and mouse endometritis,in vitro and in vivo inflammation models were established.The Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway and its downstream inflammatory factors were detected by enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qRT-PCR),western blot(WB),and immunofluorescence techniques.The results from ELISA and qRT-PCR showed that catalpol dose-dependently reduced the expression of pro-inflammatory cytokines such as tumor necrosis factorα(TNF-α),interleukin(IL)-1β,and IL-6,and chemokines such as C-X-C motif chemokine ligand 8(CXCL8)and CXCL5,both in bEECs and in uterine tissue.From the experimental results of WB,qRT-PCR,and immunofluorescence,the expression of TLR4 and the phosphorylation of NF-κB p65 were markedly inhibited by catalpol compared with the LPS group.The inflammatory damage to the mouse uterus caused by LPS was greatly reduced and was accompanied by a decline in myeloperoxidase(MPO)activity.The results of this study suggest that catalpol can exert an anti-inflammatory impact on LPS-induced bEECs and mouse endometritis by inhibiting inflammation and activation of the TLR4/NF-κB signaling pathway.
基金Project supported by the National High-Tech R & D Program (863) of China (No. 2007AA091805)and the National Natural Science Foundation of China (Nos. 30972284 and 30871944)
文摘Ds-echinoside A (DSEA),a non-sulfated triterpene glycoside,was isolated from the sea cucumber Pearsonothuria graeffei.In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion,migration,invasion,and angiogenesis.In this study,we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2,with a half-maximal inhibitory concentration (IC50) of 2.65 μmol/L,and suppressed Hep G2 cell adhesion,migration,and invasion in a dose-dependent manner.DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane (CAM) assay in vivo.Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 (MMP-9),which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis.DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1),an important regulator of MMP-9 activation.From the results of Western blotting,the expressions of nuclear factor-kappa B (NF-κB) and vascular endothelial growth factor (VEGF) were found to be remarkably reduced by DSEA.These findings suggest that DSEA exhibits a significant antimetastatic activity through the specific inhibition of NF-κB-dependent MMP-9 and VEGF expressions.
文摘目的:观察柴胡疏肝散对非酒精性脂肪性肝炎(NASH)大鼠肝脏内质网应激(ERS)相关IRE-1α/NF-κB通路的影响,探讨其防治NASH的可能作用机制。方法:采用高脂饮食12周建立NASH大鼠模型,同时以6g·kg-1·d-1的柴胡疏肝散干预12周,油红O和HE染色光镜观察肝组织脂肪变和炎性反应程度;计算NAFLD活动度(NAS);生化法检测血清ALT、AST和肝组织TG、GHOL;Realtime-PCR和Western blot技术检测肝组织GRP78、IRE-1α、NF-κB m RNA和蛋白或磷酸化水平,ELISA检测血清和肝组织中TNF-α、IL-1β水平。结果:模型组和柴胡疏肝散组大鼠肝组织均出现了脂肪变和炎性坏死。与正常组比较,模型组大鼠NAS显著增高(P<0.01),血清ALT、AST水平和肝组织TG、CHOL含量显著升高(P<0.01),肝组织GRP78、IRE-1α、NF-κB m RNA表达显著增强(P<0.01),GRP78蛋白表达和IRE-1α、NF-κB m RNA磷酸化水平显著上调(P<0.01),血清及肝组织TNF-α、IL-1β含量显著增加(P<0.01)。与模型组比较,柴胡疏肝散组大鼠NAS显著降低(P<0.01),血清ALT、AST水平和肝组织TG、CHOL含量显著降低(P<0.01),肝组织GRP78、IRE-1α、NF-κB m RNA表达显著下降(P<0.05,P<0.01),GRP78蛋白表达和IRE-1α、NF-κB m RNA磷酸化水平显著下调(P<0.05),血清及肝组织TNF-α、IL-1β含量显著下降(P<0.05)。结论:ERS相关的IRE-1α/NF-κB参与高脂饮食诱导的大鼠NASH的发生发展。柴胡疏肝散可能通过调节IRE-1α/NF-κB通路抑制过度的ERS,来调节肝脏脂质代谢紊乱、减轻肝脏炎性反应,防止NASH的进展。
