Introduction: Gastroduodenal lesions are common in chronic kidney disease (CKD). They are linked to various factors including Helicobacter pylori infection (H. pylori). Few data are available in Africa on H. pylori in...Introduction: Gastroduodenal lesions are common in chronic kidney disease (CKD). They are linked to various factors including Helicobacter pylori infection (H. pylori). Few data are available in Africa on H. pylori infection and chronic kidney disease. The aim of this study was to assess the impact of H. pylori infection and to describe the gastroduodenal lesions found in patients with chronic kidney disease. Patients and Methods: A cross-sectional study was conducted, February 1<sup>st</sup> to May 31<sup>st</sup>, 2021, at the Douala General Hospital in Cameroon. We included patients with CKD classified as stages 3 to 5 according KDIGO classification, on hemodialysis or not, who agreed to participate in the study. They were matched with a “control” population including patients with normal renal function according to sex and age (ratio 1:2). Patients on antibiotics and/or proton pump inhibitors were excluded. We collected data from CKD patients and from medical records for non-CKD group. Each patient underwent an upper digestive endoscopy and identification of H. pylori using a urease rapid test. Logistic regression was used to identify independent associations for a significance level set at p Results: We included 99 patients including 33 with CKD and 66 control patients. Among patients with CKD, the predominance was male (n = 18/33 or 54.5%). The mean age was 51.2 ± 12.8 years. Arterial hypertension was the first etiology of CKD (n = 13 or 39.4%). The prevalence of H. pylori in patients with CKD was 63.6% versus 37.9% in control patients (p-value = 0.015). The main endoscopic lesions were erosive gastropathy (n = 14 or 42.4%) and erythematous gastropathy (n = 7 or 21.2%). Patients with CKD were 5 times more likely to have H. pylori infection (OR = 5.69;CI 95% 0.14 - 0.82;p = 0.017). Factors associated with H. pylori infection were chronic kidney disease (aOR = 1.02;CI 95% 0.14 - 0.82;p = 0.017) and hemodialysis (aOR = 10;CI 95% 1.08 - 91.9;p = 0.042). Conclusion: The prevalence of H. pylori infection is higher in patients with CKD. Endoscopic lesions are inflammatory. Factors associated with H. pylori infection are chronic kidney disease and hemodialysis.展开更多
Nearly half of the global population are carriers of Helicobacter pylori(H. pylori),a Gram-negative bacterium that persists in the healthy human stomach. H. pylori can be a pathogen and causes development of peptic ul...Nearly half of the global population are carriers of Helicobacter pylori(H. pylori),a Gram-negative bacterium that persists in the healthy human stomach. H. pylori can be a pathogen and causes development of peptic ulcer disease in a certain state of the macroorganism. It is well established that H. pylori infection is the main cause of chronic gastritis and peptic ulcer disease(PUD). Decontamination of the gastric mucosa with various antibiotics leads to H. pylori elimination and longer remission in this disease. However,the reasons for repeated detection of H. pylori in recurrent PUD after its successful eradication remain unclear. The reason for the redetection of H. pylori in recurrent PUD can be either reinfection or ineffective anti-Helicobacter therapy. The administration of antibacterial drugs can lead not only to the emergence of resistant strains of microorganisms,but also contribute to the conversion of H. pylori into the resting(dormant) state. The dormant forms of H. pylori have been shown to play a potential role in the development of relapses of PUD. The paper discusses morphological H. pylori forms,such as S-shaped,C-shaped,U-shaped,and coccoid ones. The authors proposes the classification of H. pylori according to its morphological forms and viability.展开更多
Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are...Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor, the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach. Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil （E. coil） BL21. Signal peptide sequences were not included. The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting. Immunoreactivity of the proteins was determined by Western blot. Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins; and apoptosis of cells was assayed with the Cell Death ELISA kit. Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H. pylori. Apoptotic AGS cells challenged by HP1286 of H. pylori 26695 were four times more than untreated cells. In addition, apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H. pylori 26695 strain. Conclusion HP1286 of H. pylori 26695 induces apoptosis of AGS cells in a time-dependent manner, however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.展开更多
Disturbances in constitutive nitric oxide synthase (cNOS) activation associated with H. pylori colonization of gastric mucosa are considered of major consequences in defining the extent of inflammatory involvement. As...Disturbances in constitutive nitric oxide synthase (cNOS) activation associated with H. pylori colonization of gastric mucosa are considered of major consequences in defining the extent of inflammatory involvement. As rapid changes in cNOS activation are linked to the enzyme phosphorylation at the specific Ser/Thr residues, we investigated the influence of H. pylori LPS and gastric hormone, ghrelin, on the processes of phosphorylation of these two critical sites in gastric mucosal cells. We show that the LPS-induced reduction in cNOS activity is reflected in the phosphorylation on Thr497, while the countering effect of ghrelin is associated with a rapid increase in cNOS phosphorylation on Ser1179. Further, we demonstrate that cNOS phosphorylation on Thr497 as well as Ser1179 displays dependence on PKCδ. However, while the LPS-induced suppression in cNOS activation shows reliance on the phosphorylation of PKCδ and PI3K on Ser, the effect of ghrelin is manifested by the increase in phosphorylation of PKCδ and PI3K on Tyr, as well as membrane translocation and phosphorylation of Akt on Ser493. Thus, our findings suggest that the LPS-induced suppression in cNOS activation is mediated by PKCδ-controlled phosphorylation of PI3K on Ser that interferes with the membrane recruitment of Akt and promotes cNOS phosphorylation on Thr497, and that ghrelin-elicited up-regulation in cNOS activation relies on the PKCδ-directed phosphorylation of PI3K on Tyr that stimulates the membrane localization of Akt and enhances cNOS phosphorylation on Ser1179.展开更多
Disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) isozyme systems, manifested by the excessive NO and prostaglandin (PGE2) generation, are well-recognized features of gastric mucosal inflammatory re...Disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) isozyme systems, manifested by the excessive NO and prostaglandin (PGE2) generation, are well-recognized features of gastric mucosal inflammatory responses to H. pylori infection. In this study, we report that H. pylori LPS-induced enhancement in gastric mucosal inducible (i) iNOS expression and COX-2 activation was accompanied by the impairment in constitutive (c) cNOS phosphorylation, up-regulation in the inhibitory κB kinase-β (IKKβ) activation and the increase in the transcriptional factor, NF-κB, nuclear translocation. Further, we show that abrogation of cNOS control over NF-κB activation has lead to induction of iNOS expression and COX-2 activation through S-nitrosylation. Moreover, we demonstrate that the modulatory effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in the increase in Src/Akt-dependent cNOS activation through phosphorylation and the suppression of IKK-β activity through cNOS-mediated IKK-β protein S-nitrosylation. As a result, ghrelin exerted the inhibitory effect on NF-κB nuclear translocation, thus causing the repression of iNOS gene induction and the inhibition in COX-2 activation through iNOS-dependent S-nitrosylation. Our findings point to cNOS activation as a pivotal element in the signaling cascade by which ghrelin exerts modulatory control over proinflammatory events triggered in gastric mucosa by H. pylori infection.展开更多
Background: Diabetes means the blood glucose, which is too high or too low. With Type 2 DM, the more common type, the body does not make or use insulin well. In patients with DM, Helicobacter pylori is one of the most...Background: Diabetes means the blood glucose, which is too high or too low. With Type 2 DM, the more common type, the body does not make or use insulin well. In patients with DM, Helicobacter pylori is one of the most common infections worldwide. Available data on the possible association between H. pylori infection and DM are contradictory. There are a few studies in the Middle East, and this study is the pioneer study, in the Medical Services Clinics in Gaza strip. Aims: This study was conducted to reveal the prevalence of H. pylori infection, malnutrition, insulin resistance among T2DM patients, to describe the dietary requirements of T2DM patients, finally to evaluate the current information about diet, and lifestyle in the prevention of H. pylori, and malnutrition. Methodology: A cross-sectional study was conducted in the Medical Services Clinics in Gaza Strip, and there were 129 patients included in this study. Data were collected through hematological information and structured interview questionnaire. Results: Highly significant percentage of H. pylori (70%) among the DM patients includes in the study, but not indicates any significant association between gender and H. pylori status. Conclusion: H. pylori patients should update their sugar level values in the record, and should get exercise and diet plan for every meal.展开更多
BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen o...BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen on the biological behaviors of human biliary epithelium mucosa remain unknown. The present study aimed to clarify their effects on the proliferation, apoptosis, migration and oxidative DNA damage of a human intrahepatic biliary epithelial cell (HIBEC) line in vitro. METHODS: HIBECs were co -cultured with 17 beta-estradiol (at 10(-9) mol/L, 10(-7) mol/L, and 10(-5) mol/L) and H. pylori (at MOI=0.5:1, 1:1, and 2:1) and continuously passaged until the 15th generation (approximately 45 days). Then, the following assays were performed. HIBEC proliferation was measured using the CCK-8 assay, plate clone-formation assay and by determining Ki-67 expression with immunocytochemistry; cell apoptosis and migration were investigated using Annexin-V/PI and transwell assays, respectively; and reactive oxygen species (ROS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production were detected by flow cytometry and immunofluorescence staining combined with confocal laser scanning microscopy, respectively. The results were the basis for evaluating the level of oxidative stress and the related DNA damage in HIBECs. RESULTS: HIBECs maintained a normal morphology and vitality when treated with 17 beta-estradiol (at 10(-9) mol/L) and H. pylori (at MOI=0.5:1 and 1:1). 17 beta-estradiol at 10(-7) mol/L and 10(-5) mol/L and H. pylori at MOI=2:1, by contrast, caused cell death. Compared with controls, HIBECs treated with 17 beta-estradiol (10(-9) mol/L) and H. pylori (MOI=1:1) had a higher up-regulation of proliferation, Ki-67 expression, clone formation, migration activity and the expression of ROS and 8-OHdG and exhibited a down-regulation of apoptosis. The above effects were further increased when 17 beta-estradiol and H. pylori were combined (P<0.05). CONCLUSIONS: H. pylori and 17 beta-estradiol, separately or in combination, promoted cell proliferation and suppressed apoptosis of HIBECs in vitro. The above phenomena might be related to oxidative stress and its subsequent DNA damage with H. pylori and 17 beta-estradiol.展开更多
Helicobacter pylori (H. pylori) can infect into the epithelial cell to cause benign or malignant disorders. Under stressful environment, a spiral form of H. pylori is transformed into a coccoid form. The infectivity o...Helicobacter pylori (H. pylori) can infect into the epithelial cell to cause benign or malignant disorders. Under stressful environment, a spiral form of H. pylori is transformed into a coccoid form. The infectivity of the coccoid form is still controversial. Since spiral forms are transformed into two types of coccoid forms via different mechanisms, the infectivity of the two types of coccoid forms into human gastric epithelial cell was examined. A laboratory and a clinical strain of H. pyloriv were cultured in liquid medium under different conditions to produce the two types of coccoid forms. These coccoid H. pylorisv were then co-cultured with human derived gastric epithelial cell, MKN-28. Adhesion and penetration of bacteria into MKN-28 cells were monitored by scanning-, standard transmission- and immunotransmission-electron microscopy (SEM, TEM and ITEM). We observed that both coccoid forms were able to adhere onto the surface of MKN-28 cells in agminated formation and also penetrated into the gastric epithelial cells besides the spiral form of H. pyloriv. Coccoid H. pylori is not a passive entity but can actively infect the human gastric epithelial cell.展开更多
Gastric mucosal inflammatory responses to H. pylori lipopolysaccharide (LPS), are characterized by the excessive NO and prostaglandin (PGE2) generation due to the disturbances in nitric oxide synthase (NOS) and cycloo...Gastric mucosal inflammatory responses to H. pylori lipopolysaccharide (LPS), are characterized by the excessive NO and prostaglandin (PGE2) generation due to the disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) systems. Here, we report that the LPS-induced enhancement in gastric mucosal inducible (i) iNOS) activity and up-regulation in PGE2 production was associated with the suppression in Akt kinase activity and the impairment in constitutive (c) cNOS activation. The stimulatory effect of the LPS on PGE2 production, furthermore, was susceptible to suppression by COX-2 inhibitor, NS-398, and iNOS inhibitor, 1400 W. Further, we show that the countering effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in up-regu- lation in Akt activity and the increase in cNOS activation through phosphorylation, and accompanied by the suppression in iNOS expression and the reduction in COX-2 activity associated with the loss in COX-2 protein S-nitrosylation. Moreover, the effect of ghre-lin on the LPS-induced COX-2 S-nitrosylation was subject to repression by Akt inhibition. Our findings demonstrate that induction in iNOS with H. pylori in- fection leads to COX-2 activation through S-nitro- sylation and up-regulation in PGE2 generation, and that ghrelin counters these untoward consequences of the LPS through Akt-mediated up-regulation in cNO- S activation required for the iNOS gene repression.展开更多
Background: Helicobacter pylori (H. pylori) infection has been suggested to be associated with atherosclerosis. The issue is still controversial. It is well known that abnormal lipid profile is related to atherosclero...Background: Helicobacter pylori (H. pylori) infection has been suggested to be associated with atherosclerosis. The issue is still controversial. It is well known that abnormal lipid profile is related to atherosclerosis and measurement of carotid intima-media thickness. Aim of the study: to investigate carotid intima-media thickness and lipid parameters in H. pylori-positive and -negative subjects. Materials & Methods: This study was conducted in Kurdistan Teaching center of Gastroenterology and Hepatology (KCGH) in Sulaimani city during the period of December 2012 to March 2014. One hundred dyspeptic patients with H. pylori infection and 74 apparently healthy asymptomatic volunteers with H. pylori-negative tests were enrolled in this study. Both groups were comparable in age distribution and gender. H. pylori infection (IgG & IgA) were assessed by ELISA tests, Triglyceride, total cholesterol, low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) concentrations were measured by routine enzymatic methods using commercial kits. Carotid intima-media thickness was assessed by high-resolution ultrasound. Results: The mean and maximum values of internal and common carotid intima-media thickness in H. pylori-positive subjects were significantly thicker than in H. pylori-negative subjects (p H. pylori seropositive) than in controls (seronegative subjects), total cholesterol, LDL-C and triglyceride level were found to be higher in patients than in controls (p < 0.01). Conclusions: Carotid intima-media thickness as well as all lipid values apart from HDL-C was increased in H. pylori-positive subjects. These data indicated that H. pylori infection may had a role in atherosclerotic process.展开更多
文摘Introduction: Gastroduodenal lesions are common in chronic kidney disease (CKD). They are linked to various factors including Helicobacter pylori infection (H. pylori). Few data are available in Africa on H. pylori infection and chronic kidney disease. The aim of this study was to assess the impact of H. pylori infection and to describe the gastroduodenal lesions found in patients with chronic kidney disease. Patients and Methods: A cross-sectional study was conducted, February 1<sup>st</sup> to May 31<sup>st</sup>, 2021, at the Douala General Hospital in Cameroon. We included patients with CKD classified as stages 3 to 5 according KDIGO classification, on hemodialysis or not, who agreed to participate in the study. They were matched with a “control” population including patients with normal renal function according to sex and age (ratio 1:2). Patients on antibiotics and/or proton pump inhibitors were excluded. We collected data from CKD patients and from medical records for non-CKD group. Each patient underwent an upper digestive endoscopy and identification of H. pylori using a urease rapid test. Logistic regression was used to identify independent associations for a significance level set at p Results: We included 99 patients including 33 with CKD and 66 control patients. Among patients with CKD, the predominance was male (n = 18/33 or 54.5%). The mean age was 51.2 ± 12.8 years. Arterial hypertension was the first etiology of CKD (n = 13 or 39.4%). The prevalence of H. pylori in patients with CKD was 63.6% versus 37.9% in control patients (p-value = 0.015). The main endoscopic lesions were erosive gastropathy (n = 14 or 42.4%) and erythematous gastropathy (n = 7 or 21.2%). Patients with CKD were 5 times more likely to have H. pylori infection (OR = 5.69;CI 95% 0.14 - 0.82;p = 0.017). Factors associated with H. pylori infection were chronic kidney disease (aOR = 1.02;CI 95% 0.14 - 0.82;p = 0.017) and hemodialysis (aOR = 10;CI 95% 1.08 - 91.9;p = 0.042). Conclusion: The prevalence of H. pylori infection is higher in patients with CKD. Endoscopic lesions are inflammatory. Factors associated with H. pylori infection are chronic kidney disease and hemodialysis.
文摘Nearly half of the global population are carriers of Helicobacter pylori(H. pylori),a Gram-negative bacterium that persists in the healthy human stomach. H. pylori can be a pathogen and causes development of peptic ulcer disease in a certain state of the macroorganism. It is well established that H. pylori infection is the main cause of chronic gastritis and peptic ulcer disease(PUD). Decontamination of the gastric mucosa with various antibiotics leads to H. pylori elimination and longer remission in this disease. However,the reasons for repeated detection of H. pylori in recurrent PUD after its successful eradication remain unclear. The reason for the redetection of H. pylori in recurrent PUD can be either reinfection or ineffective anti-Helicobacter therapy. The administration of antibacterial drugs can lead not only to the emergence of resistant strains of microorganisms,but also contribute to the conversion of H. pylori into the resting(dormant) state. The dormant forms of H. pylori have been shown to play a potential role in the development of relapses of PUD. The paper discusses morphological H. pylori forms,such as S-shaped,C-shaped,U-shaped,and coccoid ones. The authors proposes the classification of H. pylori according to its morphological forms and viability.
基金supported by the Major Technology Project as part of "Prevention and Control of Major Infectious Diseases including AIDS and Viral Hepatitis" (No. 2008ZX10004-002)
文摘Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor, the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach. Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil （E. coil） BL21. Signal peptide sequences were not included. The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting. Immunoreactivity of the proteins was determined by Western blot. Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins; and apoptosis of cells was assayed with the Cell Death ELISA kit. Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H. pylori. Apoptotic AGS cells challenged by HP1286 of H. pylori 26695 were four times more than untreated cells. In addition, apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H. pylori 26695 strain. Conclusion HP1286 of H. pylori 26695 induces apoptosis of AGS cells in a time-dependent manner, however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.
文摘Disturbances in constitutive nitric oxide synthase (cNOS) activation associated with H. pylori colonization of gastric mucosa are considered of major consequences in defining the extent of inflammatory involvement. As rapid changes in cNOS activation are linked to the enzyme phosphorylation at the specific Ser/Thr residues, we investigated the influence of H. pylori LPS and gastric hormone, ghrelin, on the processes of phosphorylation of these two critical sites in gastric mucosal cells. We show that the LPS-induced reduction in cNOS activity is reflected in the phosphorylation on Thr497, while the countering effect of ghrelin is associated with a rapid increase in cNOS phosphorylation on Ser1179. Further, we demonstrate that cNOS phosphorylation on Thr497 as well as Ser1179 displays dependence on PKCδ. However, while the LPS-induced suppression in cNOS activation shows reliance on the phosphorylation of PKCδ and PI3K on Ser, the effect of ghrelin is manifested by the increase in phosphorylation of PKCδ and PI3K on Tyr, as well as membrane translocation and phosphorylation of Akt on Ser493. Thus, our findings suggest that the LPS-induced suppression in cNOS activation is mediated by PKCδ-controlled phosphorylation of PI3K on Ser that interferes with the membrane recruitment of Akt and promotes cNOS phosphorylation on Thr497, and that ghrelin-elicited up-regulation in cNOS activation relies on the PKCδ-directed phosphorylation of PI3K on Tyr that stimulates the membrane localization of Akt and enhances cNOS phosphorylation on Ser1179.
文摘Disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) isozyme systems, manifested by the excessive NO and prostaglandin (PGE2) generation, are well-recognized features of gastric mucosal inflammatory responses to H. pylori infection. In this study, we report that H. pylori LPS-induced enhancement in gastric mucosal inducible (i) iNOS expression and COX-2 activation was accompanied by the impairment in constitutive (c) cNOS phosphorylation, up-regulation in the inhibitory κB kinase-β (IKKβ) activation and the increase in the transcriptional factor, NF-κB, nuclear translocation. Further, we show that abrogation of cNOS control over NF-κB activation has lead to induction of iNOS expression and COX-2 activation through S-nitrosylation. Moreover, we demonstrate that the modulatory effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in the increase in Src/Akt-dependent cNOS activation through phosphorylation and the suppression of IKK-β activity through cNOS-mediated IKK-β protein S-nitrosylation. As a result, ghrelin exerted the inhibitory effect on NF-κB nuclear translocation, thus causing the repression of iNOS gene induction and the inhibition in COX-2 activation through iNOS-dependent S-nitrosylation. Our findings point to cNOS activation as a pivotal element in the signaling cascade by which ghrelin exerts modulatory control over proinflammatory events triggered in gastric mucosa by H. pylori infection.
文摘Background: Diabetes means the blood glucose, which is too high or too low. With Type 2 DM, the more common type, the body does not make or use insulin well. In patients with DM, Helicobacter pylori is one of the most common infections worldwide. Available data on the possible association between H. pylori infection and DM are contradictory. There are a few studies in the Middle East, and this study is the pioneer study, in the Medical Services Clinics in Gaza strip. Aims: This study was conducted to reveal the prevalence of H. pylori infection, malnutrition, insulin resistance among T2DM patients, to describe the dietary requirements of T2DM patients, finally to evaluate the current information about diet, and lifestyle in the prevention of H. pylori, and malnutrition. Methodology: A cross-sectional study was conducted in the Medical Services Clinics in Gaza Strip, and there were 129 patients included in this study. Data were collected through hematological information and structured interview questionnaire. Results: Highly significant percentage of H. pylori (70%) among the DM patients includes in the study, but not indicates any significant association between gender and H. pylori status. Conclusion: H. pylori patients should update their sugar level values in the record, and should get exercise and diet plan for every meal.
基金supported by grant from the National Natural Science Foundation of China(81401932)
文摘BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen on the biological behaviors of human biliary epithelium mucosa remain unknown. The present study aimed to clarify their effects on the proliferation, apoptosis, migration and oxidative DNA damage of a human intrahepatic biliary epithelial cell (HIBEC) line in vitro. METHODS: HIBECs were co -cultured with 17 beta-estradiol (at 10(-9) mol/L, 10(-7) mol/L, and 10(-5) mol/L) and H. pylori (at MOI=0.5:1, 1:1, and 2:1) and continuously passaged until the 15th generation (approximately 45 days). Then, the following assays were performed. HIBEC proliferation was measured using the CCK-8 assay, plate clone-formation assay and by determining Ki-67 expression with immunocytochemistry; cell apoptosis and migration were investigated using Annexin-V/PI and transwell assays, respectively; and reactive oxygen species (ROS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production were detected by flow cytometry and immunofluorescence staining combined with confocal laser scanning microscopy, respectively. The results were the basis for evaluating the level of oxidative stress and the related DNA damage in HIBECs. RESULTS: HIBECs maintained a normal morphology and vitality when treated with 17 beta-estradiol (at 10(-9) mol/L) and H. pylori (at MOI=0.5:1 and 1:1). 17 beta-estradiol at 10(-7) mol/L and 10(-5) mol/L and H. pylori at MOI=2:1, by contrast, caused cell death. Compared with controls, HIBECs treated with 17 beta-estradiol (10(-9) mol/L) and H. pylori (MOI=1:1) had a higher up-regulation of proliferation, Ki-67 expression, clone formation, migration activity and the expression of ROS and 8-OHdG and exhibited a down-regulation of apoptosis. The above effects were further increased when 17 beta-estradiol and H. pylori were combined (P<0.05). CONCLUSIONS: H. pylori and 17 beta-estradiol, separately or in combination, promoted cell proliferation and suppressed apoptosis of HIBECs in vitro. The above phenomena might be related to oxidative stress and its subsequent DNA damage with H. pylori and 17 beta-estradiol.
文摘Helicobacter pylori (H. pylori) can infect into the epithelial cell to cause benign or malignant disorders. Under stressful environment, a spiral form of H. pylori is transformed into a coccoid form. The infectivity of the coccoid form is still controversial. Since spiral forms are transformed into two types of coccoid forms via different mechanisms, the infectivity of the two types of coccoid forms into human gastric epithelial cell was examined. A laboratory and a clinical strain of H. pyloriv were cultured in liquid medium under different conditions to produce the two types of coccoid forms. These coccoid H. pylorisv were then co-cultured with human derived gastric epithelial cell, MKN-28. Adhesion and penetration of bacteria into MKN-28 cells were monitored by scanning-, standard transmission- and immunotransmission-electron microscopy (SEM, TEM and ITEM). We observed that both coccoid forms were able to adhere onto the surface of MKN-28 cells in agminated formation and also penetrated into the gastric epithelial cells besides the spiral form of H. pyloriv. Coccoid H. pylori is not a passive entity but can actively infect the human gastric epithelial cell.
文摘Gastric mucosal inflammatory responses to H. pylori lipopolysaccharide (LPS), are characterized by the excessive NO and prostaglandin (PGE2) generation due to the disturbances in nitric oxide synthase (NOS) and cyclooxygenase (COX) systems. Here, we report that the LPS-induced enhancement in gastric mucosal inducible (i) iNOS) activity and up-regulation in PGE2 production was associated with the suppression in Akt kinase activity and the impairment in constitutive (c) cNOS activation. The stimulatory effect of the LPS on PGE2 production, furthermore, was susceptible to suppression by COX-2 inhibitor, NS-398, and iNOS inhibitor, 1400 W. Further, we show that the countering effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in up-regu- lation in Akt activity and the increase in cNOS activation through phosphorylation, and accompanied by the suppression in iNOS expression and the reduction in COX-2 activity associated with the loss in COX-2 protein S-nitrosylation. Moreover, the effect of ghre-lin on the LPS-induced COX-2 S-nitrosylation was subject to repression by Akt inhibition. Our findings demonstrate that induction in iNOS with H. pylori in- fection leads to COX-2 activation through S-nitro- sylation and up-regulation in PGE2 generation, and that ghrelin counters these untoward consequences of the LPS through Akt-mediated up-regulation in cNO- S activation required for the iNOS gene repression.
文摘Background: Helicobacter pylori (H. pylori) infection has been suggested to be associated with atherosclerosis. The issue is still controversial. It is well known that abnormal lipid profile is related to atherosclerosis and measurement of carotid intima-media thickness. Aim of the study: to investigate carotid intima-media thickness and lipid parameters in H. pylori-positive and -negative subjects. Materials & Methods: This study was conducted in Kurdistan Teaching center of Gastroenterology and Hepatology (KCGH) in Sulaimani city during the period of December 2012 to March 2014. One hundred dyspeptic patients with H. pylori infection and 74 apparently healthy asymptomatic volunteers with H. pylori-negative tests were enrolled in this study. Both groups were comparable in age distribution and gender. H. pylori infection (IgG & IgA) were assessed by ELISA tests, Triglyceride, total cholesterol, low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) concentrations were measured by routine enzymatic methods using commercial kits. Carotid intima-media thickness was assessed by high-resolution ultrasound. Results: The mean and maximum values of internal and common carotid intima-media thickness in H. pylori-positive subjects were significantly thicker than in H. pylori-negative subjects (p H. pylori seropositive) than in controls (seronegative subjects), total cholesterol, LDL-C and triglyceride level were found to be higher in patients than in controls (p < 0.01). Conclusions: Carotid intima-media thickness as well as all lipid values apart from HDL-C was increased in H. pylori-positive subjects. These data indicated that H. pylori infection may had a role in atherosclerotic process.
