BACKGROUND Helicobacter pylori(H.pylori)infection can cause extensive apoptosis of gastric epithelial cells,serving as a critical catalyst in the progression from chronic gastritis,gastrointestinal metaplasia,and atyp...BACKGROUND Helicobacter pylori(H.pylori)infection can cause extensive apoptosis of gastric epithelial cells,serving as a critical catalyst in the progression from chronic gastritis,gastrointestinal metaplasia,and atypical gastric hyperplasia to gastric carcinoma.Prompt eradication of H.pylori is paramount for ameliorating the pathophysiological conditions associated with chronic inflammation of the gastric mucosa and the primary prevention of gastric cancer.Acacetin,which has multifaceted pharmacological activities such as anti-cancer,anti-inflammatory,and antioxidative properties,has been extensively investigated across various domains.Nevertheless,the impact and underlying mechanisms of action of acacetin on H.pylori-infected gastric mucosal epithelial cells remain unclear.AIM To explore the defensive effects of acacetin on apoptosis in H.pylori-infected GES-1 cells and to investigate the underlying mechanisms.METHODS GES-1 cells were treated with H.pylori and acacetin in vitro.Cell viability was assessed using the CCK-8 assay,cell mortality rate via lactate dehydrogenase assay,alterations in cell migration and healing capacities through the wound healing assay,rates of apoptosis via flow cytometry and TUNEL staining,and expression levels of apoptosis-associated proteins through western blot analysis.RESULTS H.pylori infection led to decreased GES-1 cell viability,increased cell mortality,suppressed cell migration,increased rate of apoptosis,increased expressions of Bax and cle-caspase3,and decreased Bcl-2 expression.Conversely,acacetin treatment enhanced cell viability,mitigated apoptosis induced by H.pylori infection,and modulated the expression of apoptosis-regulatory proteins by upregulating Bcl-2 and downregulating Bax and cleaved caspase-3.CONCLUSION Acacetin significantly improved GES-1 cell viability and inhibited apoptosis in H.pylori-infected GES-1 cells,thereby exerting a protective effect on gastric mucosal epithelial cells.展开更多
Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 pr...Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 protein levels in all six gastric epithelial cell lines were investigated by Western blot analysis.Gene copy number was accessed and then sequenced the coding region from each mRNA in all six cell lines.The HtrA1 promoter region DNA methylation status was detected by using bisulfite sequeucing analysis.Effect of decitabine and TSA on HTRA1 expression in gastric cancer cell line was determined by RTPCR.Results:HIC analysis indicated that HtrA1 was highly expressed in normal epithelium,but dramatically down-regulated in gastric carcinoma tissues and variably expressed in tumor-adjacent tissues.HtrA1 gene expression was dramatically decreased in gastric carcinoma cells compared to nontumorigenic counterparts.The HtrA1 gene loss in any of the 4 breast cancer cell lines was not detected.Total 14 CpGs in this region were all methylated in gastric cancer cells,whereas two normal cells.GES-1 and HFI-145,were having several unmethylated cytosines in this region.HtrA1 showed as^Mr 44,000,Expression of HtrA1 protein was not observed in any of the four gastric caucer cell lines.BGC-823.MKN-45.SGC-7901and MKN-28.HtrA1 expression was observed in the HF1-145and GES-1 cell lines.Conclusions:The epigenetic silencing for HtrA1gene expression could provide a possible strategy for re-activating Htrt1 gene expression in gastric cancer cells.thus facilitating further investigation of HtrA1's role in chemotherapy.展开更多
Flammulina velutipes(F.velutipes)polysaccharides were modified by ultrasound at the rated power of 150 W and 900 W.The monosaccharide composition,ultraviolet-visible,and Fourier transform infrared spectral characteris...Flammulina velutipes(F.velutipes)polysaccharides were modified by ultrasound at the rated power of 150 W and 900 W.The monosaccharide composition,ultraviolet-visible,and Fourier transform infrared spectral characteristics of F.velutipes polysaccharides(FVP)and their ultrasonic modification products(U-FVPs)were determined.The protective effects of FVP and U-FVPs on human gastric mucosal cells GES-1 were confi rmed for the first time.The mole ratios of glucose and galactose were decreased and the mole ratio of mannose was increased after ultrasonic modification.Compared with the original FVP and the FVP modifi ed by ultrasound of 150 W(U-FVP1),the FVP modifi ed by ultrasound of 900 W(U-FVP2)could better prevent ethanol-induced damage to GES-1 cells.With increasing ultrasound intensity,the protective effect of FVPs on GES-1 cells was significantly enhanced by more effective prevention of intracellular reactive oxygen species(ROS)production and more promotion of expression of triglyceride factor 2(TFF2),prostaglandin E2(PGE2),epidermal growth factor(EGF),and transforming growth factorβ1(TGF-β1)mRNA.The ultrasonic modifi cation might be an effective way to develop novel F.velutipes polysaccharides that could effectively resist the gastric injury caused by excessive alcohol consumption.展开更多
基金Supported by the Doctoral Research Initiation Fund of Affiliated Hospital of Southwest Medical University,No.21037.
文摘BACKGROUND Helicobacter pylori(H.pylori)infection can cause extensive apoptosis of gastric epithelial cells,serving as a critical catalyst in the progression from chronic gastritis,gastrointestinal metaplasia,and atypical gastric hyperplasia to gastric carcinoma.Prompt eradication of H.pylori is paramount for ameliorating the pathophysiological conditions associated with chronic inflammation of the gastric mucosa and the primary prevention of gastric cancer.Acacetin,which has multifaceted pharmacological activities such as anti-cancer,anti-inflammatory,and antioxidative properties,has been extensively investigated across various domains.Nevertheless,the impact and underlying mechanisms of action of acacetin on H.pylori-infected gastric mucosal epithelial cells remain unclear.AIM To explore the defensive effects of acacetin on apoptosis in H.pylori-infected GES-1 cells and to investigate the underlying mechanisms.METHODS GES-1 cells were treated with H.pylori and acacetin in vitro.Cell viability was assessed using the CCK-8 assay,cell mortality rate via lactate dehydrogenase assay,alterations in cell migration and healing capacities through the wound healing assay,rates of apoptosis via flow cytometry and TUNEL staining,and expression levels of apoptosis-associated proteins through western blot analysis.RESULTS H.pylori infection led to decreased GES-1 cell viability,increased cell mortality,suppressed cell migration,increased rate of apoptosis,increased expressions of Bax and cle-caspase3,and decreased Bcl-2 expression.Conversely,acacetin treatment enhanced cell viability,mitigated apoptosis induced by H.pylori infection,and modulated the expression of apoptosis-regulatory proteins by upregulating Bcl-2 and downregulating Bax and cleaved caspase-3.CONCLUSION Acacetin significantly improved GES-1 cell viability and inhibited apoptosis in H.pylori-infected GES-1 cells,thereby exerting a protective effect on gastric mucosal epithelial cells.
基金supported by Natural Science and Technology Fund of Hubei Povince(012726334)
文摘Objective:To explore HtrA1 gene expression aud its regulation in human gastric cancers.Methods:The HtrA1 mRNA levels were examined by QPCR analysis and coufirmed its expression with Northern blot analysis.The HtrA1 protein levels in all six gastric epithelial cell lines were investigated by Western blot analysis.Gene copy number was accessed and then sequenced the coding region from each mRNA in all six cell lines.The HtrA1 promoter region DNA methylation status was detected by using bisulfite sequeucing analysis.Effect of decitabine and TSA on HTRA1 expression in gastric cancer cell line was determined by RTPCR.Results:HIC analysis indicated that HtrA1 was highly expressed in normal epithelium,but dramatically down-regulated in gastric carcinoma tissues and variably expressed in tumor-adjacent tissues.HtrA1 gene expression was dramatically decreased in gastric carcinoma cells compared to nontumorigenic counterparts.The HtrA1 gene loss in any of the 4 breast cancer cell lines was not detected.Total 14 CpGs in this region were all methylated in gastric cancer cells,whereas two normal cells.GES-1 and HFI-145,were having several unmethylated cytosines in this region.HtrA1 showed as^Mr 44,000,Expression of HtrA1 protein was not observed in any of the four gastric caucer cell lines.BGC-823.MKN-45.SGC-7901and MKN-28.HtrA1 expression was observed in the HF1-145and GES-1 cell lines.Conclusions:The epigenetic silencing for HtrA1gene expression could provide a possible strategy for re-activating Htrt1 gene expression in gastric cancer cells.thus facilitating further investigation of HtrA1's role in chemotherapy.
基金supported by the Special Funds for Scientific and Technological Achievement Transformation Project in Jiangsu Province(BA2021062).
文摘Flammulina velutipes(F.velutipes)polysaccharides were modified by ultrasound at the rated power of 150 W and 900 W.The monosaccharide composition,ultraviolet-visible,and Fourier transform infrared spectral characteristics of F.velutipes polysaccharides(FVP)and their ultrasonic modification products(U-FVPs)were determined.The protective effects of FVP and U-FVPs on human gastric mucosal cells GES-1 were confi rmed for the first time.The mole ratios of glucose and galactose were decreased and the mole ratio of mannose was increased after ultrasonic modification.Compared with the original FVP and the FVP modifi ed by ultrasound of 150 W(U-FVP1),the FVP modifi ed by ultrasound of 900 W(U-FVP2)could better prevent ethanol-induced damage to GES-1 cells.With increasing ultrasound intensity,the protective effect of FVPs on GES-1 cells was significantly enhanced by more effective prevention of intracellular reactive oxygen species(ROS)production and more promotion of expression of triglyceride factor 2(TFF2),prostaglandin E2(PGE2),epidermal growth factor(EGF),and transforming growth factorβ1(TGF-β1)mRNA.The ultrasonic modifi cation might be an effective way to develop novel F.velutipes polysaccharides that could effectively resist the gastric injury caused by excessive alcohol consumption.
文摘[目的]研究腹水草(Veronicastrum axillare,V.axillare)含药血清对乙醇损伤的人胃上皮细胞(gastric epithelial cells-1,GES-1)的保护作用及其对环腺苷酸依赖性激酶(protein kinase A,PKA)、环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)、水通道蛋白1(aquaporin 1,AQP1)表达水平的调控作用。[方法]20只雄性SD大鼠随机分为5组(n=4),连续灌胃给药14d,制备正常组(生理盐水20 m L·kg-1)、雷尼替丁组(0.027g·kg-1)以及V.axillare低、中、高剂量组(0.7、1.4、2.8 g·kg-1)含药血清。MTT法检测体外培养的GES-1细胞的活性,研究V.axillare含药血清及PKA抑制剂H-89(25μmol·m L-1)预处理对5%乙醇诱导GES-1细胞活力的影响。荧光定量RT-PCR检测PKA、CREB及AQP1 m RNA的表达水平,Western blot检测AQP1蛋白表达水平。[结果]模型组和H-89组GES-1细胞活性低于正常组,差异有统计学意义(P<0.01);乙醇造模处理显著上调PKA、CREB及下调AQP1的表达水平,差异有统计学意义(P<0.01);H-89处理显著抑制PKA和CREB m RNA表达水平(P<0.01);加入各剂量V.axillare含药血清不能显著改善H-89对PKA、CREB和AQP1 m RNA表达水平的抑制作用。V.axillare中、高剂量含药血清细胞活性高于模型组,PKA、CREB m RNA表达水平低于模型组,差异有统计学意义(P<0.01),而V.axillare中、低剂量含药血清AQP1表达水平高于模型组,差异有统计学意义(P<0.01)。[结论]V.axillare含药血清显著减轻乙醇对GES-1细胞的损伤,对GES-1细胞具有保护作用,其保护机制与下调PKA、CREB及上调AQP1的表达有关。
