Immunoregulatory effects of leptin on inflammatory bowel disease / 中华微生物学和免疫学杂志

Inflammatory bowel disease (IBD) is a chronic and recurrent disease characterized by chronic inflammation of the gastrointestinal tract. The quality of life of patients with IBD is seriously affected. The pathogenesis of IBD is complex, among which immune factors are considered to be the predominant factor. Leptin is a hormone derived from adipocytes and recent studies have shown that it is involved in the regulation of immune cells and inflammatory signaling pathways. This review summarized the pathogenesis of IBD, the immunoregulatory mechanism of leptin and research progress in immune modulators, introduced the potential effects of leptin on the regulation of immunological homeostasis in IBD and its potential roles in the etiology and pathogenesis of IBD, and discussed a possible immunotherapy method to treat IBD through leptin antagonist to reduce the inflammatory response and inhibit the inflammatory signaling pathways.

Clinical study on Fuzheng Zhuyu Xiehuo Decoction combine with conventional western medicine therapy in the treatment of acute pancreatitis of intermingled blood stasis-toxin syndrome / 国际中医中药杂志

Objective:To explore the efficacy of Fuzheng Zhuyu Xiehuo Decoction for the patients with acute pancreatitis (AP) of intermingled blood stasis-toxin syndrome and its influence on peripheral blood inflammatory factors and microcirculation indicators.Methods:A total of 100 patients with AP, admitted to department of spleen and stomach diseases of the First Affiliated Hospital of Hunan University of Chinese Medicine and department of gastroenterology of the Central Hospital of Shaoyang, who met the inclusion criteria between March 2019 and March 2020, were divided into two groups according to the random number table method, with 50 in each group. The control group was given conventional western medicine, while the observation group was treated with Fuzheng Zhuyu Xiehuo Decoction on the basis of the control group. The TCM syndromes were scored before and after treatment, and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) was used to evaluate the severity of the disease, and ELISA was adopted to detect the levels of IL-6, IL-8, TNF-α and thromboxane A2 (TXA2), prostaglandin I 2 (PGI 2) and platelet activating factor (PAF). The abdominal pain, abdominal distension, fever, gastrointestinal function recovery time and hospital stay were observed and the adverse events were recorded. Results:The total effective rate was 96.0% (48/50) in the observation group and that of the control group was 84.0% (42/50) ( χ2=4.00, P=0.045). The scores of abdominal pain, abdominal distension, fever and nausea and vomiting in observation group were significantly lower than those in the control group ( t=7.07, 7.06, 11.47, 10.30, all Ps<0.01), and the recovery times of abdominal pain, abdominal distension, fever and gastrointestinal function and hospital stay in the observation group were significantly shorter than those in the control group ( t=4.52, 4.90, 6.27, 6.55, 7.12, all Ps<0.01). After treatment, the levels of serum IL-6 [(30.15±7.04) μg/L vs. (42.37±8.29) μg/L, t=7.95], IL-8 [(39.36±8.11) μg/L vs. (50.36±10.47) μg/L, t=5.87], TNF-α [(106.28±21.04) μg/L vs. (153.45±30.23) μg/L, t=9.06] in the observation group were significantly lower than those in the control group ( P<0.01). The serum TXA2 [(223.68±40.15) ng/L vs. (257.11±50.32) ng/L, t=3.67] and PAF [(74.86±15.37) ng/L vs. (85.53±15.26) ng/L, t=3.48] in the observation group were significantly lower than those in the control group ( P<0.01) while the level of PGI 2 [(91.43±17.45) ng/L vs. (76.49±15.13) ng/L, t=4.57] in the observation group was significantly higher than those in the control group ( P<0.01). Conclusion:Fuzheng Zhuyu Xiehuo Decoction combined with western medicine can improve clinical symptoms and blood microcirculation status, relieve inflammatory response and enhance clinical efficacy of patients with AP of intermingled blood stasis-toxin syndrome.

Genetic and expression changes in TNF-α as a risk factor for rheumatoid arthritis pathogenesis in northeast India

Antitumour necrosis factor-alpha (TNF-α) therapy is used as a clinical intervention for rheumatoid arthritis (RA) but differences exist in response to the treatment which makes the candidature of the screening of TNF-α alteration(s) at genetic and expression levels an important agenda prior to treatment. This study aims to determine the associative role of TNF-α –308G/A polymorphism and differential expression of TNF-α in the pathogenesis of RA. A case–control study where a total of 126 RA patients were enrolled based on ACR-EULAR (2010) criteria, along with 160 community matched age and sex controls over a period of three years. The differential expression level of TNF-α mRNA and protein level was studied and TNF-α –308G/A polymorphism was screened by T-ARMS PCR assay. All statistical analysis was performed using SPSS software. mRNA expression level of TNF-α was upregulated in RA cases (avg. 15.85 ± 9.52 fold) compared to control. TNF-α protein level was found to be higher in RA cases (28.62±7.17 pg/mL) compared to control (23.14±6.91 pg/mL). TNF-α –308 variant GA genotype was higher in RA (46.03%) than in control (25%). The presence of TNF-α –308 variant A allele was associated with increased risk of RA susceptibility (odds ratio (OR) = 2.559 at 95% confidence interval (CI), P< 0.001) but not severity (OR = 1.617 at 95% CI, P = 0.571). The presence of –308 variant genotype was associated with a higher TNF-α mRNA and protein expression. The presence of TNF-α –308A allele is associated with increased risk of RA susceptibility and differential TNF-α expression, and has prognostic significance. Association of higher TNF-α pro-inflammatory cytokine levels with northeast Indian patients makes them suitable subjects for anti-TNF-α therapy.

Role of toll like-receptor 2 in inflammatory activity of macrophage infected with a recombinant BCG expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum

Objective: To investigate the role of toll-like receptor 2 (TLR2) in inflammatory activity of macrophage infected with the recombinant Mycobacterium bovis bacillus Calmette-Guerin (rBCG). Methods: Mouse macrophage cell line J774A.1 was infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG) and rBCG cultures for 48 h in the presence or absence of 10 μg/mL of TLR2 inhibitor. Untreated macrophages were used as a negative control while lipopolysaccharide-stimulated macrophages were used as a positive control. The ability of the macrophage to engulf the BCG and rBCG in the absence or presence of TLR2 inhibitor was assessed using a phagocytic assay, while the production of inflammatory cytokines and nitric oxide by the infected macrophages was evaluated using ELISA and Griess reagent method, while the expression of the inducible nitric oxide synthase was determined using Western blot analysis. Results: The results showed that blocking TLR2 function reduced the phagocytic activity, nitric oxide production and proinflammatory cytokine secretion such as TNF- α, IL-1 β and IL-12p40 as well as inducible nitric oxide synthase expression in the infected macrophages. These data showed the importance of TLR2 in the activation of macrophages following BCG and rBCG infections. Conclusions: Through exploring the immunological mechanism which underlies the protection conferred by the candidate vaccine, this study will improve our understanding of the vaccine candidate's mechanism to protect the host from malaria infection.

Role of toll like-receptor 2 in inflammatory activity of macrophage infected with a recombinant BCG expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum

Objective: To investigate the role of toll-like receptor 2 (TLR2) in inflammatory activity of macrophage infected with the recombinant Mycobacterium bovis bacillus Calmette-Guerin (rBCG). Methods: Mouse macrophage cell line J774A.1 was infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG) and rBCG cultures for 48 h in the presence or absence of 10 μg/mL of TLR2 inhibitor. Untreated macrophages were used as a negative control while lipopolysaccharide-stimulated macrophages were used as a positive control. The ability of the macrophage to engulf the BCG and rBCG in the absence or presence of TLR2 inhibitor was assessed using a phagocytic assay, while the production of inflammatory cytokines and nitric oxide by the infected macrophages was evaluated using ELISA and Griess reagent method, while the expression of the inducible nitric oxide synthase was determined using Western blot analysis. Results: The results showed that blocking TLR2 function reduced the phagocytic activity, nitric oxide production and proinflammatory cytokine secretion such as TNF-α, IL-1β and IL-12p40 as well as inducible nitric oxide synthase expression in the infected macrophages. These data showed the importance of TLR2 in the activation of macrophages following BCG and rBCG infections. Conclusions: Through exploring the immunological mechanism which underlies the protection conferred by the candidate vaccine, this study will improve our understanding of the vaccine candidate's mechanism to protect the host from malaria infection.

Mechanism of elevated proinflammatory cytokines in aqueous humor of patients with primary angle-closure glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

Glaucomais a group of diseases characterized by optic atrophy and visual field defect.In China,primary angle closure glaucoma (PACG) is the most common type of glaucoma.The mechanism of glaucoma has many theories,such as mechanical theory and vascular theory.Recent researches found that inflammation may be involved in the pathogenesis of glaucoma.A variety of proinflammatory cytokines significantly increased in aqueous humor of patients with PACG.In this study,we summarized the methods for the detection of aqueous humor,and analyzed the mechanism of the increasing of proinflammatory cytokines in the aqueous humor of patients with PACG.

Advances in research on the central mechanism of heart failure / 西安交通大学学报(医学版)

Heart failure is a complex clinical syndrome,and impaired filling or ejection disorders result from any ventricular structure or dysfunction can cause heart failure.The prevalence of heart failure in adult populations in developed countries has reached 1% to 2%,while the prevalence in elderly people over 70 years of age has increased to ≥ 10%. With the population aging and the prevalence of coronary heart disease increased, the prevalence of heart failure has increased,becoming a disable and fatal disease.The changes of the central nervous system hormones such as renin-angiotensin system (RAS),inflammatory cytokines (proinflammatory cytokines,PIC),and reactive oxygen species (ROS)may be closely related to increased central activity in heart failure,which can significantly alter the activities of peripheral sympathetic nerves. Constant sympathetic nervous activity is an important cause of development of heart failure,so reducing the sympathetic excitability of heart failure is regarded as one of the focuses of treatment and research.This paper focuses on the influence of central neurohormone on heart failure and possible central mechanism.

Analgesic effect and central mechanisms of CQ prescription on cancer invasion induced mirror image pain in model mice / 中国中药杂志

This study aimed to analyze the analgesic effect and related central mechanisms of CQ prescription on cancer invasion induced mirror image pain (CIIMIP)in model mice.In the study, male BALB/c mice were randomly divided into normal group, operation control group (injected with 0.2 mL inactivated S180 sarcoma cell sap), model group (injected with 0.2 mL S180 sarcoma cell sap on the right leg near the greater trochanter of femur) and CQ prescription low dose group (intraperitoneally injected with CQ prescription 100 mg•kg⁻¹ on the basis of model mice), CQ prescription middle dose group (intraperitoneally injected with CQ prescription 150 mg•kg⁻¹ on the basis of model mice), and CQ prescription high dose group (intraperitoneally injected with CQ prescription 200 mg•kg⁻¹ on the basis of model mice). Mechanical withdraw threshold (MWT) of the mirror image lateral hind paws were evaluated by Von Frey hairs before modeling and after surgery. The levels of glutamate (Glu), gamma aminobutyric acid (GABA), glycine (Gly), and taurine (Tau) in the L3-L5 spinal cord were measured by the high performance liquid chromatography-fluorescence detector (HPLC-FLD); AimPlex detection technology with multiple factors was used to detect the levels of regulated on activation in normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein (MCP-3) in the L3-L5 spinal cord. Then we observed the influence of GABAa receptor antagonist (Bicuculline) on analgesic effect of CQ prescription.The results indicated that CQ prescription could remarkably increase MWT of model mice(P<0.01, P<0.05), decrease the level of Glu(P<0.01, P<0.05), improve the levels of GABA, Gly, Tau(P<0.01, P<0.05), lower the ratio of Glu/GABA(P<0.01, P<0.05), and reduce the levels of RANTES, MCP-3(P<0.05) in the L3-L5 spinal cord, and GABAa receptor antagonist significantly blocked the analgesic effect of CQ prescription at two time points(P<0.05).This study showed that CQ prescription had significant analgesic effect on CIIMIP model mice, and its mechanism was associated with regulating the balance between excitability amino acid(EAA) and inhibitory amino acid (IAA) transmitters in central nervous system, partially activating GABAa receptor, and reducing the release of RANTES and MCP-3 in the spinal cord.

Advances in research on the central mechanism of hypertension / 西安交通大学学报(医学版)

ABSTRACT:Hypertension,the first risk factor for stroke and coronary heart disease in the Chinese population, seriously endangers people’s health.At present,China has more than 270 million people with hypertension and an annual increase rate of 1 0 million people.Then how to improve prevention and treatment of hypertension has become an urgent need to solve major medical and social problems.In the past,research on hypertension mainly focused on the peripheral area,while recent research has shown that the central regulation plays an important role in the development of hypertension. Hypothalamic paraventricular nucleus (PVN ), which plays a key role in maintaining cardiovascular activity, can directly control the sympathetic preganglionic neurons and regulate peripheral sympathetic nerve activity,thus being closely related to the development of hypertension.Research in recent years shows that the comprehensive effects of proinflammatory cytokines (PIC ),reactive oxygen species (ROS),renin-angiotensin system (RAS),neurotransmitter (NT)and nuclear factorκB (NF-κB)are involved in the pathogenesis of hypertension.However,it is unclear how these neurohormones in PVN are activated,how they interact with each other and what role they play in the regulatory mechanism of hypertension.Therefore,the key focus of this research is to explore the impact of activated neurohormones in PVN on hypertension.This study will provide new content for the study on hypertension.

Preinduced intestinal HSP70 improves visceral hypersensitivity and abnormal intestinal motility in PI-IBS mouse model

Objective: To investigate the impact of the preinduced intestinal heat shock protein 70 (HSP70) on the visceral hypersensitivity and abnormal intestinal motility in a post-infectious irritable bowel syndrome (PI-IBS) mouse model. Methods: Eighty-four female C57BL/6 mice were randomly assigned to four groups: control group (n = 21) and induction + PI-IBS group (n = 21), PI-IBS group (n = 21) and induction group (n = 21). The mice in PI-IBS group were infected in vivo with Trichinella spiralis by oral administration. The visceral hypersensitivity and intestinal motility were evaluated respectively with abdominal withdrawal reflex and colon transportation test. The intestinal HSP70 protein and mRNA level was measured by Western blot and real-time PCR. Meanwhile, the intestinal proinflammatory cytokines IL-10 and TNF-α level was detected by ELISA. Results: Compared with their counterparts in PI-IBS group, the animals in the Induction + PI-IBS group show significantly increased intestinal level of HSP70 and obviously ameliorative clinical figures, including abdominal withdrawal reflex score, intestine transportation time and Bristol scores (P < 0.05). Meanwhile, the intestinal post-inflammatory cytokines remarkably changed, including increased IL-10 level and decreased TNF-α level (P < 0.05). Conclusions: Intestinal HSP70 may play a potential protective role through improving the imbalance between the intestinal post-inflammatory and anti-inflammatory cytokines in PI-IBS.

Preinduced intestinal HSP70 improves visceral hypersensitivity and abnormal intestinal motility in PI-IBS mouse model

OBJECTIVE@#To investigate the impact of the preinduced intestinal heat shock protein 70 (HSP70) on the visceral hypersensitivity and abnormal intestinal motility in a post-infectious irritable bowel syndrome (PI-IBS) mouse model.@*METHODS@#Eighty-four female C57BL/6 mice were randomly assigned to four groups: control group (n = 21) and induction + PI-IBS group (n = 21), PI-IBS group (n = 21) and induction group (n = 21). The mice in PI-IBS group were infected in vivo with Trichinella spiralis by oral administration. The visceral hypersensitivity and intestinal motility were evaluated respectively with abdominal withdrawal reflex and colon transportation test. The intestinal HSP70 protein and mRNA level was measured by Western blot and real-time PCR. Meanwhile, the intestinal proinflammatory cytokines IL-10 and TNF-α level was detected by ELISA.@*RESULTS@#Compared with their counterparts in PI-IBS group, the animals in the Induction + PI-IBS group show significantly increased intestinal level of HSP70 and obviously ameliorative clinical figures, including abdominal withdrawal reflex score, intestine transportation time and Bristol scores (P < 0.05). Meanwhile, the intestinal post-inflammatory cytokines remarkably changed, including increased IL-10 level and decreased TNF-α level (P < 0.05).@*CONCLUSIONS@#Intestinal HSP70 may play a potential protective role through improving the imbalance between the intestinal post-inflammatory and anti-inflammatory cytokines in PI-IBS.

Acetyl Eburicoic Acid from Laetiporus sulphureus var. miniatus Suppresses Inflammation in Murine Macrophage RAW 264.7 Cells

The basidiomycete Laetiporus sulphureus var. miniatus belongs to the Aphyllophorales, Polyporaceae, and grows on the needleleaf tree. The fruiting bodies of Laetiporus species are known to produce N-methylated tyramine derivatives, polysaccharides, and various lanostane triterpenoids. As part of our ongoing effort to discover biologically active compounds from wood-rotting fungi, an anti-inflammatory triterpene, LSM-H7, has been isolated from the fruiting body of L. sulphureus var. miniatus and identified as acetyl eburicoic acid. LSM-H7 dose-dependently inhibited the NO production in RAW 264.7 cells without any cytotoxicity at the tested concentrations. Furthermore it suppressed the production of proinflammatory cytokines, mainly inducible nitric oxide synthase, cyclooxygenase-2, interleukin (IL)-1beta, IL-6 and tumor necrosis factor alpha, when compared with glyceraldehyde 3-phosphate dehydrogenase. These data suggest that LSM-H7 is a crucial component for the anti-inflammatory activity of L. sulphureus var. miniatus.

Human Rhinovirus-induced Proinflammatory Cytokine and Interferon-beta Responses in Nasal Epithelial Cells From Chronic Rhinosinusitis Patients

PURPOSE: Asthma exacerbation from human rhinovirus (HRV) infection is associated with deficient antiviral interferon (IFN) secretion. Although chronic rhinosinusitis (CRS), an inflammatory upper airway disease, is closely linked to asthma, IFN-beta responses to HRV infections in human nasal epithelial cells (HNECs) from CRS patients remain to be studied. We evaluated inflammatory and antiviral responses to HRV infection in HNECs from CRS patients. METHODS: HNECs, isolated from turbinate tissue of 13 patients with CRS and 14 non-CRS controls, were infected with HRV16 for 4 hours. The HRV titer, LDH activity, production of proinflammatory cytokines and IFN-beta proteins, and expression levels of RIG-I and MDA5 mRNA were assessed at 8, 24, and 48 hours after HRV16 infection. RESULTS: The reduction in viral titer was slightly delayed in the CRS group compared to the non-CRS control group. IL-6 and IL-8 were significantly increased to a similar extent in both groups after HRV infection. In the control group, IFN-beta production and MDA5 mRNA expression were significantly increased at 8 and 24 hours after HRV16 infection, respectively. By contrast, in the CRS group, IFN-beta was not induced by HRV infection; however, HRV-induced MDA5 mRNA expression was increased, but the increase was slightly delayed compared to the non-CRS control group. The RIG-I mRNA level was not significantly increased by HRV16 infection in either group. CONCLUSIONS: HRV-induced secretion of proinflammatory cytokines in CRS patients was not different from that in the non-CRS controls. However, reductions in viral titer, IFN-beta secretion, and MDA5 mRNA expression in response to HRV infection in CRS patients were slightly impaired compared to those in the controls, suggesting that HRV clearance in CRS patients might be slightly deficient.

Migraine & paediatric obesity: a plausible link.

Obesity and migraine are both highly prevalent disorders in the general population, influenced by genetic and environmental risk factors. In recent studies, obesity was found to be a strong risk factor for transformed migraine and, among migraineurs, obesity was associated with frequent headaches and higher disability scores. Suggested mechanisms included: (i) obesity as a pro-inflammatory state may be associated with neurovascular inflammation in patients with migraine; (ii) elevated levels of plasma calcitonin gene-related peptide (CGRP) in obese individuals may play a role as an important post-synaptic mediator of trigeminovascular inflammation in migraine; (iii) dismodulation in the hypothalamic neuropeptide, orexin, in obese persons may be associated with increased susceptibility to neurogenic inflammation causing migraine attacks; and (iv) leptin and adiponectin can activate proinflammatory cytokine release that is involved in the pathogenesis of migraine. In addition, both conditions are associated with psychiatric co-morbidities, such as depression and anxiety, that can further increase headache frequency and disability. Therefore, the effect of obesity on migraine outcome is important. Weight and BMI should be measured and calculated in all children presenting with migraine, and weight control should be a part of the treatment.

Anti-inflammatory Effect of Plocamium telfairiae Extract in LPS-stimulated Bone Marrow-derived Macrophages and Dendritic Cells

Marine algae are rich sources of various biologically active compounds with potential pharmaceutical properties. In the present study, we investigated the inhibitory effects of Plocamium telfairiae extract (PTE) on proinflammatory cytokine production in bone marrow-derived macrophage (BMDMs) and dendritic cells (BMDCs). PTE pre-treatment in LPS-stimulated BMDMs and BMDCs showed a strong inhibition on interleukin (IL)-12 p40, IL-6, and tumor necrosis factor (TNF)-alpha production as compared to non-treated controls. PTE pre-treatment showed significant inhibition on phosphorylation of mitogen-activated protein kinases and degradation of inhibitor of kappa B (IkappaBalpha). Taken together, these results suggest that PTE may have potential anti-inflammatory property and hence, warrant further studies concerning the potentials of PTE for medicinal purpose.

The role of Toll-like receptor signal in inflammatory response in neonatal sepsis / 中国小儿急救医学

Objective To investigate the role of signal transduction of Toll-like receptors (TLRs) in inflammatory response of neonatal sepsis.Methods Twenty children with neonatal sepsis and 16 health neonates were studied.Real-time PCR were used to evaluate the levels of TLR1 ～ TLR10,myeloid differentiation protein 2 (MD-2),myeloid differentiation factor 88 (MyD88),interleukin (IL)-1β,IL-6,IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression in peripheral blood mononuclear cells.Expressions of proinflammatory cytokines (IL-1β,IL-6,IL-8,TNF-α) were measured by ELISA.Results (1) Compared with control group,the mRNA levels of TLR2,TLR4 in neonatal sepsis group were up-regulated significantly (TLR2:55.16±12.78 vs 9.53 ± 3.73,P ＜ 0.01 ;TLR4:125.22 ±30.64 vs 23.17 ± 5.78,P ＜0.01),the differences were not significant as to other TLRs.(2) Transcription levels of MD-2 and MyD88 were significantly up-regulated in neonatal sepsis group (MD-2:376.83 ± 62.16 vs 12.92 ± 2.54,P ＜ 0.01 ; MyD88:11.97 ±2.48 vs 2.77 ±0.59,P ＜0.01).(3) Expressions of proinflammatory cytokines (IL-1β,IL-6,IL-8,TNF-α) in neonatal sepsis group were higher than those of control group [PCR:(IL-1β:21.72 ± 5.56 vs 5.69 ± 1.26,P ＜0.01 ;IL-6:71.39 ± 18.34 vs 9.65 ±2.13,P ＜0.01 ;IL-8:29.39 ±6.72 vs 8.72 ± 1.95,P＜0.01;TNF-α:65.42 ± 16.95 vs 12.33 ±3.45,P ＜0.01).ELISA:(IL-1β:2 977.36 ±653.97 vs 480.52 ± 120.36,P ＜ 0.01 ; IL-6:3 143.82 ± 775.08 vs 393.78 ± 96.55,P ＜ 0.01 ; IL-8:2 510.78 ± 686.77 vs 276.91 ±72.46,P ＜0.01 ;TNF-α:3 582.24 ± 876.13 vs 1 233.68 ± 289.39,P ＜ 0.01)].Conclusion Abnormal activation of TLRs and higher expressions of proinflammatory cytokines in neonatal sepsis,suggesting that aberrant activation of TLRs may be one of the initiating factors of immune aberrance in neonatal sepsis.

Genetic factors in the pathogenesis of gastroesophageal reflux disease.

Multiple factors play a role in the pathogenesis of gastroesophageal reflux disease (GERD). Two landmark studies showing higher concordance of disease in monozygotic than dizygotic twin pairs suggested the role of host genetic factors in its pathogenesis. Recent studies have shown that genetic polymorphism in genes influencing host’s inflammatory response, drug metabolism, cell cycle regulation, xenobiotic pathways, DNA repair, mutagenesis, esophageal sensory function and gene silencing are associated with risk of GERD and its sequelae—Barrett’s esophagus and esophageal adenocarcinoma. However, more studies on larger sample size are needed before reaching a definite conclusion on the role of an individual gene.

GLP-1 Can Protect Proinflammatory Cytokines Induced Beta Cell Apoptosis through the Ubiquitination

BACKGROUND: Proinflammatory cytokines are one of the causes of diabetes mellitus. However, the exact molecular mechanism by which proinflammatory cytokines induce beta-cell death remains to be clearly elucidated. Glucagon-like peptide-1 (GLP-1) affects the stimulation of insulin secretion and the preservation of beta-cells. Additionally, it may exert an antiapoptotic effect on beta cells; however, the mechanism underlying this effect has yet to be demonstrated. Therefore, we investigated the protective effects of GLP-1 in endoplasmic reticulum (ER)-mediated beta-cell apoptosis using proinflammatory cytokines. METHODS: To induce ER stress, hamster insulin-secreting tumor (HIT)-T15 cells were treated using a mixture of cytokines. Apoptosis was evaluated via MTT assay, Hoechst 33342 staining, and annexin/propidium iodide (PI) flow cytometry. The mRNA and protein expression levels of ER stress-related molecules were determined via PCR and Western blotting, respectively. Nitric oxide was measured with Griess reagent. The levels of inducible nitric oxide synthase (iNOS) mRNA and protein were analyzed via real-time PCR and Western blot, respectively. iNOS protein degradation was evaluated via immunoprecipitation. We pretreated HIT-T15 cells with exendin (Ex)-4 for 1 hour prior to the induction of stress. RESULTS: We determined that Ex-4 exerted a protective effect through nitric oxide and the modulation of ER stress-related molecules (glucose-regulated protein [GRP]78, GRP94, and CCAAT/enhancer-binding protein homologous protein [CHOP]) and that Ex-4 stimulates iNOS protein degradation via the ubiquitination pathway. Additionally, Ex-4 also induced the recovery of insulin2 mRNA expression in beta cells. CONCLUSION: The results of this study indicate that GLP-1 may protect beta cells against apoptosis through the ubiquitination pathway.

Proinflammatory Cytokine and Nitric Oxide Production by Human Macrophages Stimulated with Trichomonas vaginalis

Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.

Proinflammatory Cytokine and Nitric Oxide Production by Human Macrophages Stimulated with Trichomonas vaginalis

Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.