Hepatocellular carcinoma(HCC)is a leading cause of death worldwide.Current therapies are effective for HCC patients with early disease,but many patients suffer recurrence after surgery and have a poor response to chem...Hepatocellular carcinoma(HCC)is a leading cause of death worldwide.Current therapies are effective for HCC patients with early disease,but many patients suffer recurrence after surgery and have a poor response to chemotherapy.Therefore,new therapeutic targets are needed.We analyzed gene expression profiles between HCC tissues and normal adjacent tissues from public databases and found that the expression of genes involved in lipid metabolism was significantly different.The analysis showed that AKR1C3 was upregulated in tumors,and high AKR1C3 expression was associated with a poorer prognosis in HCC patients.In vitro,assays demonstrated that the knockdown of AKR1C3 or the addition of the AKR1C3 inhibitor indomethacin suppressed the growth and colony formation of HCC cell lines.Knockdown of AKR1C3 in Huh7 cells reduced tumor growth in vivo.To explore the mechanism,we performed pathway enrichment analysis,and the results linked the expression of AKR1C3 with prostaglandin F2 alpha(PGF2a)downstream target genes.Suppression of AKR1C3 activity reduced the production of PGF2a,and supplementation with PGF2a restored the growth of indomethacin-treated Huh7 cells.Knockdown of the PGF receptor(PTGFR)and treatment with a PTGFR inhibitor significantly reduced HCC growth.We showed that indomethacin potentiated the sensitivity of Huh7 cells to sorafenib.In summary,our results indicate that AKR1C3 upregulation may promote HCC growth by promoting the production of PGF2α,and suppression of PTGFR limited HCC growth.Therefore,targeting the AKR1C3-PGF2a-PTGFR axis may be a new strategy for the treatment of HCC.展开更多
AIM： Grapefruit-seed extract （GSE） containing flavonoids, possesses antibacterial and antioxidative properties but whether it influences the gastric defense mechanism and gastroprotection against ethanol- and stres...AIM： Grapefruit-seed extract （GSE） containing flavonoids, possesses antibacterial and antioxidative properties but whether it influences the gastric defense mechanism and gastroprotection against ethanol- and stress-induced gastric lesions remains unknown. METHODS： We compared the effects of GSE on gastric mucosal lesions induced in rats by topical application of 100% ethanol or 3.5 h of water immersion and restraint stress （WRS） with or without （A） inhibition of cyclooxygenase （COX）-1 activity by indomethacin and rofecoxib, the selective COX-2 inhibitor, （B） suppression of NO-synthase with L-NNA （20 mg/kg ip）, and （C） inactivation by capsaicin （125 mg/kg sc） of sensory nerves with or without intragastric （ig） pretreatment with GSE applied 30 min prior to ethanol or WRS. One hour after ethanol and 3.5 h after the end of WRS, the number and area of gastric lesions were measured by planimetry, the gastric blood flow （GBF） was assessed by H2-gas clearance technique and plasma gastrin levels and the gastric mucosal generation of PGE2, superoxide dismutase （SOD） activity and malonyldialdehyde （MDA） concentration, as an index of lipid peroxidation were determined. RESULTS： Ethanol and WRS caused gastric lesions accompanied by the significant fall in the GBF and SOD activity and the rise in the mucosal MDA content. Pretreatment with GSE （8-64 mg/kg i g） dose- dependently attenuated gastric lesions induced by 100% ethanol and WRS; the dose reducing these lesions by 50% （ID50） was 25 and 36 mg/kg, respectively, and this protective effect was similar to that obtained with methyl PGE2 analog （5 μg/kg i g）. GSE significantly raised the GBF, mucosal generation of PGE2, SOD activity and plasma gastrin levels while attenuating IVlDA content. Inhibition of PGE2 generation with indomethacin or rofecoxib and suppression of NO synthase by L-NNA or capsaicin denervation reversed the GSE-induced protection and the accompanying hyperemia. Cotreatment of exogenous calcitonine gene-related peptide （CGRP） with GSE restored the protection and accompanying hyperemic effects of GSE in rats with capsaicin denervation. CONCLUSION： GSE exerts a potent gastroprotective activity against ethanol and WRS-induced gastric lesions via an increase in endogenous PG generation, suppression of lipid peroxidation and hyperemia possibly mediated by NO and CGRP released from sensory nerves.展开更多
AIM：To explore the role of prostaglandin F2α（PGF2α） on pacemaker activity in interstitial cells of Cajal（ICC）from mouse small intestine. METHODS：In this study,effects of PGF2αin the cultured ICC cells were in...AIM：To explore the role of prostaglandin F2α（PGF2α） on pacemaker activity in interstitial cells of Cajal（ICC）from mouse small intestine. METHODS：In this study,effects of PGF2αin the cultured ICC cells were investigated with patch clamp technology combined with Ca 2＋ image analysis. RESULTS：Externally applied PGF2α（10μmol/L）produced membrane depolarization in current-clamp mode and increased tonic inward pacemaker currents in voltage-clamp mode.The application of flufenamic acid（a non-selective cation channel inhibitor）or niflumic acid（aCl channel inhibitor）abolished the generation of pacemaker currents but only flufenamic acid inhibited the PGF2α-induced tonic inward currents.In addition,the tonic inward currents induced by PGF2αwere not inhibited by intracellular application of 5’-[-thio]diphosphate trilithium salt.Pretreatment with Ca 2＋ free solution, U-73122,an active phospholipase C inhibitor,and thapsigargin,a Ca 2＋ -ATPase inhibitor in endoplasmic reticulum,abolished the generation of pacemaker currents and suppressed the PGF2α-induced tonic inward currents.However,chelerythrine or calphostin C,protein kinase C inhibitors,did not block the PGF2α-induced effects on pacemaker currents.When recording intracellular Ca 2＋ （[Ca 2＋ ]i）concentration using fluo-3/AM,PGF2α broadly increased the spontaneous[Ca 2＋ ]i oscillations. CONCLUSION：These results suggest that PGF2αcan modulate pacemaker activity of ICC by acting non-selective action channels through phospholipase C-dependent pathway via[Ca2＋]i regulation展开更多
Hyperhomocysteinemia and oxidative stress may be strongly linked to hypertension, atherosclerosis and other cardiovascular diseases. The present study was performed to investigate possible relationships among plasma t...Hyperhomocysteinemia and oxidative stress may be strongly linked to hypertension, atherosclerosis and other cardiovascular diseases. The present study was performed to investigate possible relationships among plasma total homocysteine, plasma 8-iso-prostaglandin F2α (8-isoPG F2α: an index of oxidative stress), and membrane fluidity (a reciprocal value of membrane microviscosity) in hypertension. We measured the membrane fluidity of red blood cells (RBCs) in hypertensive and normotensive men using an electron spin resonance (ESR) and spin-labeling method. Membrane fluidity of RBCs was significantly decreased in hypertensive men compared with normotensive men. Plasma total homocysteine levels were significantly higher in hypertensive men than in normotensive men, and correlated with plasma 8-isoPG F2α. In contrast, plasma nitric oxide (NO)-metabolites (an index of endothelial function) were lower in hypertensive men than in normotensive men. The reduced membrane fluidity of RBCs was associated with increased total homocysteine and plasma 8-isoPG F2α levels and decreased plasma NO-metabolite levels. Multivariate regression analysis showed that, after adjusting for general risk factors, plasma total homocysteine and 8-isoPG F2α were significant determinants of membrane fluidity of RBCs, respectively. These results suggest that hyperhomocysteinemia and oxidative stress with endothelial dysfunction might have a close correlation with impaired rheologic behavior of RBCs and circulatory disorders in hypertensive men.展开更多
目的 8 异前列腺素F2α(8 iso PGF2α)是一种敏感、特异性反映缺血 再灌注后氧自由基增加的生 化指标。抗氧化剂N 乙酰半胱氨酸(NAC)具有清除氧自由基的作用。本研究旨在分析实验性心肌缺血大鼠血浆 与心肌8 iso PGF2α相关性以及...目的 8 异前列腺素F2α(8 iso PGF2α)是一种敏感、特异性反映缺血 再灌注后氧自由基增加的生 化指标。抗氧化剂N 乙酰半胱氨酸(NAC)具有清除氧自由基的作用。本研究旨在分析实验性心肌缺血大鼠血浆 与心肌8 iso PGF2α相关性以及NAC的治疗效果,探讨血浆8 iso PGF2α反映心肌氧自由基损伤程度的可能性和 NAC的干预效果。方法 45只雄性成年Wistar大鼠随机分为3组(每组15只):对照组、缺血组和NAC组。缺血 组和NAC组腹腔注射垂体后叶素(20U/kg)制成大鼠急性心肌缺血模型,以心电图上ST段的抬高作为心肌缺血 的指标。对照组仅腹腔注射生理盐水。NAC组缺血前2周开始用NAC(每日0.1g/kg)灌胃,共3周。应用ELISA 方法测定各组大鼠血浆及心肌组织8 iso PGF2α含量。结果 缺血组大鼠的血浆和心肌组织含量分别为(187.1± 45.8)pg/mL和(259.3±47.5)pg/g,明显高于正常对照组(60.4±13.7)pg/mL和(88.6±16.9)pg/g (P<0.01);NAC组的血浆和心肌组织8 iso PGF2α含量为(88.2±16.4)pg/mL和(109.4±24.7)pg/g明显低于 缺血组(P<0.01)。血浆与心肌8 iso PGF2α水平相关(r=0.856,P<0.01)。与正常组比较,缺血组的心电图 ST段明显抬高(心肌缺血45min时抬高最为明显,达0.34±0.05mV)(P<0.05);展开更多
基金National Yang Ming Chiao Tung University Far Eastern Memorial Hospital Joint Research Programs(NYCU-FEMH 109DN03,110DN06,111DN04,112DN05).
文摘Hepatocellular carcinoma(HCC)is a leading cause of death worldwide.Current therapies are effective for HCC patients with early disease,but many patients suffer recurrence after surgery and have a poor response to chemotherapy.Therefore,new therapeutic targets are needed.We analyzed gene expression profiles between HCC tissues and normal adjacent tissues from public databases and found that the expression of genes involved in lipid metabolism was significantly different.The analysis showed that AKR1C3 was upregulated in tumors,and high AKR1C3 expression was associated with a poorer prognosis in HCC patients.In vitro,assays demonstrated that the knockdown of AKR1C3 or the addition of the AKR1C3 inhibitor indomethacin suppressed the growth and colony formation of HCC cell lines.Knockdown of AKR1C3 in Huh7 cells reduced tumor growth in vivo.To explore the mechanism,we performed pathway enrichment analysis,and the results linked the expression of AKR1C3 with prostaglandin F2 alpha(PGF2a)downstream target genes.Suppression of AKR1C3 activity reduced the production of PGF2a,and supplementation with PGF2a restored the growth of indomethacin-treated Huh7 cells.Knockdown of the PGF receptor(PTGFR)and treatment with a PTGFR inhibitor significantly reduced HCC growth.We showed that indomethacin potentiated the sensitivity of Huh7 cells to sorafenib.In summary,our results indicate that AKR1C3 upregulation may promote HCC growth by promoting the production of PGF2α,and suppression of PTGFR limited HCC growth.Therefore,targeting the AKR1C3-PGF2a-PTGFR axis may be a new strategy for the treatment of HCC.
文摘AIM： Grapefruit-seed extract （GSE） containing flavonoids, possesses antibacterial and antioxidative properties but whether it influences the gastric defense mechanism and gastroprotection against ethanol- and stress-induced gastric lesions remains unknown. METHODS： We compared the effects of GSE on gastric mucosal lesions induced in rats by topical application of 100% ethanol or 3.5 h of water immersion and restraint stress （WRS） with or without （A） inhibition of cyclooxygenase （COX）-1 activity by indomethacin and rofecoxib, the selective COX-2 inhibitor, （B） suppression of NO-synthase with L-NNA （20 mg/kg ip）, and （C） inactivation by capsaicin （125 mg/kg sc） of sensory nerves with or without intragastric （ig） pretreatment with GSE applied 30 min prior to ethanol or WRS. One hour after ethanol and 3.5 h after the end of WRS, the number and area of gastric lesions were measured by planimetry, the gastric blood flow （GBF） was assessed by H2-gas clearance technique and plasma gastrin levels and the gastric mucosal generation of PGE2, superoxide dismutase （SOD） activity and malonyldialdehyde （MDA） concentration, as an index of lipid peroxidation were determined. RESULTS： Ethanol and WRS caused gastric lesions accompanied by the significant fall in the GBF and SOD activity and the rise in the mucosal MDA content. Pretreatment with GSE （8-64 mg/kg i g） dose- dependently attenuated gastric lesions induced by 100% ethanol and WRS; the dose reducing these lesions by 50% （ID50） was 25 and 36 mg/kg, respectively, and this protective effect was similar to that obtained with methyl PGE2 analog （5 μg/kg i g）. GSE significantly raised the GBF, mucosal generation of PGE2, SOD activity and plasma gastrin levels while attenuating IVlDA content. Inhibition of PGE2 generation with indomethacin or rofecoxib and suppression of NO synthase by L-NNA or capsaicin denervation reversed the GSE-induced protection and the accompanying hyperemia. Cotreatment of exogenous calcitonine gene-related peptide （CGRP） with GSE restored the protection and accompanying hyperemic effects of GSE in rats with capsaicin denervation. CONCLUSION： GSE exerts a potent gastroprotective activity against ethanol and WRS-induced gastric lesions via an increase in endogenous PG generation, suppression of lipid peroxidation and hyperemia possibly mediated by NO and CGRP released from sensory nerves.
基金Supported by Research Fund from Chosun Hospital 2008
文摘AIM：To explore the role of prostaglandin F2α（PGF2α） on pacemaker activity in interstitial cells of Cajal（ICC）from mouse small intestine. METHODS：In this study,effects of PGF2αin the cultured ICC cells were investigated with patch clamp technology combined with Ca 2＋ image analysis. RESULTS：Externally applied PGF2α（10μmol/L）produced membrane depolarization in current-clamp mode and increased tonic inward pacemaker currents in voltage-clamp mode.The application of flufenamic acid（a non-selective cation channel inhibitor）or niflumic acid（aCl channel inhibitor）abolished the generation of pacemaker currents but only flufenamic acid inhibited the PGF2α-induced tonic inward currents.In addition,the tonic inward currents induced by PGF2αwere not inhibited by intracellular application of 5’-[-thio]diphosphate trilithium salt.Pretreatment with Ca 2＋ free solution, U-73122,an active phospholipase C inhibitor,and thapsigargin,a Ca 2＋ -ATPase inhibitor in endoplasmic reticulum,abolished the generation of pacemaker currents and suppressed the PGF2α-induced tonic inward currents.However,chelerythrine or calphostin C,protein kinase C inhibitors,did not block the PGF2α-induced effects on pacemaker currents.When recording intracellular Ca 2＋ （[Ca 2＋ ]i）concentration using fluo-3/AM,PGF2α broadly increased the spontaneous[Ca 2＋ ]i oscillations. CONCLUSION：These results suggest that PGF2αcan modulate pacemaker activity of ICC by acting non-selective action channels through phospholipase C-dependent pathway via[Ca2＋]i regulation
文摘Hyperhomocysteinemia and oxidative stress may be strongly linked to hypertension, atherosclerosis and other cardiovascular diseases. The present study was performed to investigate possible relationships among plasma total homocysteine, plasma 8-iso-prostaglandin F2α (8-isoPG F2α: an index of oxidative stress), and membrane fluidity (a reciprocal value of membrane microviscosity) in hypertension. We measured the membrane fluidity of red blood cells (RBCs) in hypertensive and normotensive men using an electron spin resonance (ESR) and spin-labeling method. Membrane fluidity of RBCs was significantly decreased in hypertensive men compared with normotensive men. Plasma total homocysteine levels were significantly higher in hypertensive men than in normotensive men, and correlated with plasma 8-isoPG F2α. In contrast, plasma nitric oxide (NO)-metabolites (an index of endothelial function) were lower in hypertensive men than in normotensive men. The reduced membrane fluidity of RBCs was associated with increased total homocysteine and plasma 8-isoPG F2α levels and decreased plasma NO-metabolite levels. Multivariate regression analysis showed that, after adjusting for general risk factors, plasma total homocysteine and 8-isoPG F2α were significant determinants of membrane fluidity of RBCs, respectively. These results suggest that hyperhomocysteinemia and oxidative stress with endothelial dysfunction might have a close correlation with impaired rheologic behavior of RBCs and circulatory disorders in hypertensive men.
文摘目的 8 异前列腺素F2α(8 iso PGF2α)是一种敏感、特异性反映缺血 再灌注后氧自由基增加的生 化指标。抗氧化剂N 乙酰半胱氨酸(NAC)具有清除氧自由基的作用。本研究旨在分析实验性心肌缺血大鼠血浆 与心肌8 iso PGF2α相关性以及NAC的治疗效果,探讨血浆8 iso PGF2α反映心肌氧自由基损伤程度的可能性和 NAC的干预效果。方法 45只雄性成年Wistar大鼠随机分为3组(每组15只):对照组、缺血组和NAC组。缺血 组和NAC组腹腔注射垂体后叶素(20U/kg)制成大鼠急性心肌缺血模型,以心电图上ST段的抬高作为心肌缺血 的指标。对照组仅腹腔注射生理盐水。NAC组缺血前2周开始用NAC(每日0.1g/kg)灌胃,共3周。应用ELISA 方法测定各组大鼠血浆及心肌组织8 iso PGF2α含量。结果 缺血组大鼠的血浆和心肌组织含量分别为(187.1± 45.8)pg/mL和(259.3±47.5)pg/g,明显高于正常对照组(60.4±13.7)pg/mL和(88.6±16.9)pg/g (P<0.01);NAC组的血浆和心肌组织8 iso PGF2α含量为(88.2±16.4)pg/mL和(109.4±24.7)pg/g明显低于 缺血组(P<0.01)。血浆与心肌8 iso PGF2α水平相关(r=0.856,P<0.01)。与正常组比较,缺血组的心电图 ST段明显抬高(心肌缺血45min时抬高最为明显,达0.34±0.05mV)(P<0.05);
