Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory p...Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.展开更多
AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transfor...AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.展开更多
Objective:To explore the relationship between the expression of TLR4 and TIRAP and sepsis severity.Methods:The study selected 30 healthy examinees as the control group and 53 patients with sepsis as the observation gr...Objective:To explore the relationship between the expression of TLR4 and TIRAP and sepsis severity.Methods:The study selected 30 healthy examinees as the control group and 53 patients with sepsis as the observation group.The patients in the observation group were assessed by Acute Physiology and Chronic Health Evaluation Ⅱ(APACHE Ⅱ).40 patients with APACHE Ⅱ score≤20,were classified into the low-score sepsis group;13 patients with APACHE Ⅱ score>20,were classified into the high-score sepsis group.The levels of TLR4 and TIRAP in venous serum were detected in all subjects by enzyme-linked immunosorbent assay(ELISA).Results:The levels of TLR4 and TIRAP in serum were 0.886±0.058 ng/ml and 5.216±0.410 ng/ml in the control group;2.253±0.379 ng/ml and 9.540±2.294 ng/ml in the low-score sepsis group;4.494±0.709 ng/ml and 19.206±1.755 ng/ml in the high-score sepsis group.The observation group(low-score and high-score sepsis groups)was significantly different from the control group(p=.000),and the low-score sepsis group was significantly different from the high-score sepsis group(p=.000).With APACHE II score of 20 as the cut-off point,the low-score sepsis group was consisted of low-risk patients and the high-score group was consisted of the high-risk,which can indicate the sepsis severity.According to Pearson’s correlation analysis,the level of TLR4 was positively correlated with sepsis severity(r=0.931,p<.05),the level of TIRAP was also positively correlated with the sepsis severity(r=0.972,p<.05);the level of TLR4 was positively correlated with the level of TIRAP(r=0.936,p<.05).Conclusions:The levels of TLR4 and TIRAP in septic patients can be used to predict and determine the severity of sepsis.展开更多
OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided...OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided into the control(n=10)and CAG groups(n=55).CAG model rats were further divided into five groups:model(n=10),vitacoenzyme(n=10),low-dose QHY(n=10),medium-dose QHY(n=10),and high-dose QHY groups(n=10).We analyzed histopathological changes using hematoxylin and eosin staining and measured interleukin(IL)-6 and IL-8 levels in serum using enzyme-linked immunosorbent assay(ELISA)(Boster Bio,Pleasanton,USA).In addition,gastrin(GAS),pepsinogen I(PGI),and PGII expressions were evaluated using ELISA.The protein and m RNA expression of toll-like receptor 4(TLR4)and toll or interleukin-1 receptor domaincontaining adaptor inducing interferon-β(TRIF)was detected by Western blotting and quantitative reverse transcription-polymerase chain reaction,respectively.RESULTS:Our results revealed that histopathological changes in CAG model rates could be restored by low-,medium-,and high-dose QHY.The changes in GAS and PGI/II expression demonstrated that QHY improved CAG.Serum IL-6 and IL-levels were decreased by QHY administration.TLR4 and TRIF were upregulated at the m RNA and protein levels in the model group but downregulated by QHY administration.CONCLUSION:We concluded that QHY could effectively improve the histopathological changes of the gastric mucosa induced by CAG in rats.The therapeutic mechanism of QHY may be related to inhibition of the inflammatory factors IL-6 and IL-8 and suppression of TLR4/TRIF m RNA and protein expression.展开更多
BACKGROUND The prevention and treatment of Hirschsprung-associated enterocolitis(HAEC)is a serious challenge in pediatric surgery.Exploring the mechanism of HAEC is conducive to the prevention of this disease.AIM To e...BACKGROUND The prevention and treatment of Hirschsprung-associated enterocolitis(HAEC)is a serious challenge in pediatric surgery.Exploring the mechanism of HAEC is conducive to the prevention of this disease.AIM To explore the possible mechanism of glycyrrhizic acid(GA)and its therapeutic effect on HAEC.METHODS We developed a model of enteritis induced by trinitrobenzenesulfonic acid(TNBS)in zebrafish,and treated it with different concentrations of GA.We analyzed the effect of GA on the phenotype and inflammation of zebrafish.RESULTS After treatment with TNBS,the area of the intestinal lumen in zebrafish was significantly increased,but the number of goblet cells in the intestinal lumen was significantly reduced,but these did not increase the mortality of zebrafish,indicating that the zebrafish enteritis model was successfully developed.Different concentrations of GA protected zebrafish with enteritis.In particular,high concentrations of GA were important for the prevention and control of HAEC because it significantly reduced the intestinal luminal area,increased the number of goblet cells in the intestinal lumen,and reduced the levels of interleukin(IL)-1βand IL-8.CONCLUSION GA significantly reduced the intestinal luminal area,increased the number of intestinal goblet cells,and decreased IL-1βand IL-8 in zebrafish,and is important for prevention and control of HAEC.展开更多
There is a major transformation in gene expression between mature B cells （including follicular, marginal zone, and germinal center cells） and antibody secreting cells （ASCs）, i.e. , ASCs, （including plasma blas...There is a major transformation in gene expression between mature B cells （including follicular, marginal zone, and germinal center cells） and antibody secreting cells （ASCs）, i.e. , ASCs, （including plasma blasts, splenic plasma cells, and long-lived bone marrow plasma cells）. This signifcant change-over occurs to accommodate the massive amount of secretory-specific immunoglobulin that ASCs make and the export processes itself. It is well known that there is an up-regulation of a small number of ASC-specific transcription factors Prdm1 （B-lymphocyte-induced maturation protein 1）, interferon regulatory factor 4, and Xbp1, and the reciprocal down-regulation of Pax5, Bcl6 and Bach2, which maintain the B cell program. Less well appreciated are the major alterations in transcription elongation and RNA proce-ssing occurring between B cells and ASCs. The three ELL family members ELL1, 2 and 3 have different protein sequences and potentially distinct cellular roles in transcription elongation. ELL1 is involved in DNA repair and small RNAs while ELL3 was previously described as either testis or stem-cell specifc. After B cell stimulation to ASCs, ELL3 levels fall precipitously while ELL1 falls off slightly. ELL2 is induced at least 10-fold in ASCs relative to B cells. All of these changes cause the RNA Polymerase Ⅱ in ASCs to acquire different properties, leading to differences in RNA processing and histone modifcations.展开更多
Background:Traumatic brain edema(TBE)is caused by a specific water channel mediated by membrane aquaporins.Aquaporin-4(AQP4)plays an especially important role in this process,but the relationship between AQP4 and TBE ...Background:Traumatic brain edema(TBE)is caused by a specific water channel mediated by membrane aquaporins.Aquaporin-4(AQP4)plays an especially important role in this process,but the relationship between AQP4 and TBE remains unclear.The purpose of this study was to explore expression of AQP4 in the hippocampus after traumatic brain injury(TBI),as well as the effect of brain edema on skeletal protein and its function in hippocampal neurons.Methods:The adult male Wistar rats we divided into a sham group and a TBI group,the latter of which was further divided into 1,3,6,12,24 and 72 hours(h)and 15 days(d)post injury subgroups.A proper TBI model was established,and brain edema was assessed in each group by water content.We measured the abundance of various proteins,including hypoxia inducible factor-1α(HIF-1α),AQP4,microtubule-associated protein 2(MAP2),tau-5 protein,phosphorylated level of TAU,synaptophysin,cyclic adenosine monophosphate response element binding protein(CREB),phosphorylated CREB and general control nonrepressed 2,in each group.Hippocampal neurons and spatial memory test were analyzed in different time points.Results:Compared with that in the sham group,the level of AQP4 in hippocampal neurons began to significantly increase at 1 h post TBI and then decreased at 15 d post TBI.During this time frame,AQP4 level peaked at 12 and 72 h,and these peaks were closely correlated with high brain water content.HIF-1αdisplayed a similar trend.Conversely,levels of MAP2 began to decrease at 1 h post TBI and then increase at 15 d post TBI.In addition,the most severe brain edema in rats was found at 24 h post TBI,with neuronal loss and hippocampal dendritic spine injury.Compared to those in the sham group,rats in the TBI groups had significantly prolonged latency and significantly shortened exploration time.Conclusions:AQP4 level was closely correlated with severity of brain edema,and abnormal levels thereof aggravated such severity after TBI.展开更多
基金supported by the National Natural Science Foundation of ChinaNos.81971047 (to WTL) and 82073910 (to XFW)+2 种基金the Natural Science Foundation of Jiangsu Province,No.BK20191253 (to XFW)Key R&D Program (Social Development) Project of Jiangsu Province,No.BE2019 732 (to WTL)Jiangsu Province Hospital (the First Affiliated Hospital of Nanjing Medical University) Clinical Capacity Enhancement Project,No.JSPH-511B2018-8 (to YBP)。
文摘Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.
文摘AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.
文摘Objective:To explore the relationship between the expression of TLR4 and TIRAP and sepsis severity.Methods:The study selected 30 healthy examinees as the control group and 53 patients with sepsis as the observation group.The patients in the observation group were assessed by Acute Physiology and Chronic Health Evaluation Ⅱ(APACHE Ⅱ).40 patients with APACHE Ⅱ score≤20,were classified into the low-score sepsis group;13 patients with APACHE Ⅱ score>20,were classified into the high-score sepsis group.The levels of TLR4 and TIRAP in venous serum were detected in all subjects by enzyme-linked immunosorbent assay(ELISA).Results:The levels of TLR4 and TIRAP in serum were 0.886±0.058 ng/ml and 5.216±0.410 ng/ml in the control group;2.253±0.379 ng/ml and 9.540±2.294 ng/ml in the low-score sepsis group;4.494±0.709 ng/ml and 19.206±1.755 ng/ml in the high-score sepsis group.The observation group(low-score and high-score sepsis groups)was significantly different from the control group(p=.000),and the low-score sepsis group was significantly different from the high-score sepsis group(p=.000).With APACHE II score of 20 as the cut-off point,the low-score sepsis group was consisted of low-risk patients and the high-score group was consisted of the high-risk,which can indicate the sepsis severity.According to Pearson’s correlation analysis,the level of TLR4 was positively correlated with sepsis severity(r=0.931,p<.05),the level of TIRAP was also positively correlated with the sepsis severity(r=0.972,p<.05);the level of TLR4 was positively correlated with the level of TIRAP(r=0.936,p<.05).Conclusions:The levels of TLR4 and TIRAP in septic patients can be used to predict and determine the severity of sepsis.
基金Supported by Natural Science Foundation of Fujian Province(Based on NLRP3 Inflammatory Body/Caspase-1-mediated Gastric Epithelial Cell Death to Explore the Molecular Mechanism of Qinghua Decoction in the Treatment of Chronic Atrophic Gastritis,No.2020J01253)。
文摘OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided into the control(n=10)and CAG groups(n=55).CAG model rats were further divided into five groups:model(n=10),vitacoenzyme(n=10),low-dose QHY(n=10),medium-dose QHY(n=10),and high-dose QHY groups(n=10).We analyzed histopathological changes using hematoxylin and eosin staining and measured interleukin(IL)-6 and IL-8 levels in serum using enzyme-linked immunosorbent assay(ELISA)(Boster Bio,Pleasanton,USA).In addition,gastrin(GAS),pepsinogen I(PGI),and PGII expressions were evaluated using ELISA.The protein and m RNA expression of toll-like receptor 4(TLR4)and toll or interleukin-1 receptor domaincontaining adaptor inducing interferon-β(TRIF)was detected by Western blotting and quantitative reverse transcription-polymerase chain reaction,respectively.RESULTS:Our results revealed that histopathological changes in CAG model rates could be restored by low-,medium-,and high-dose QHY.The changes in GAS and PGI/II expression demonstrated that QHY improved CAG.Serum IL-6 and IL-levels were decreased by QHY administration.TLR4 and TRIF were upregulated at the m RNA and protein levels in the model group but downregulated by QHY administration.CONCLUSION:We concluded that QHY could effectively improve the histopathological changes of the gastric mucosa induced by CAG in rats.The therapeutic mechanism of QHY may be related to inhibition of the inflammatory factors IL-6 and IL-8 and suppression of TLR4/TRIF m RNA and protein expression.
基金Joint Funds for the Innovation of Science and Technology,Fujian Province,No.2020Y9139Startup Fund for Scientific Research,Fujian Medical University,No.2019QH1141.
文摘BACKGROUND The prevention and treatment of Hirschsprung-associated enterocolitis(HAEC)is a serious challenge in pediatric surgery.Exploring the mechanism of HAEC is conducive to the prevention of this disease.AIM To explore the possible mechanism of glycyrrhizic acid(GA)and its therapeutic effect on HAEC.METHODS We developed a model of enteritis induced by trinitrobenzenesulfonic acid(TNBS)in zebrafish,and treated it with different concentrations of GA.We analyzed the effect of GA on the phenotype and inflammation of zebrafish.RESULTS After treatment with TNBS,the area of the intestinal lumen in zebrafish was significantly increased,but the number of goblet cells in the intestinal lumen was significantly reduced,but these did not increase the mortality of zebrafish,indicating that the zebrafish enteritis model was successfully developed.Different concentrations of GA protected zebrafish with enteritis.In particular,high concentrations of GA were important for the prevention and control of HAEC because it significantly reduced the intestinal luminal area,increased the number of goblet cells in the intestinal lumen,and reduced the levels of interleukin(IL)-1βand IL-8.CONCLUSION GA significantly reduced the intestinal luminal area,increased the number of intestinal goblet cells,and decreased IL-1βand IL-8 in zebrafish,and is important for prevention and control of HAEC.
基金Supported by The National Science Foundation grant MCB-08-42725National Institutes of Health shared resources Grant No.P30CA047904 to the University of Pittsburgh Cancer Instituteinternal funding from the School of Medicine and Department of Immunology
文摘There is a major transformation in gene expression between mature B cells （including follicular, marginal zone, and germinal center cells） and antibody secreting cells （ASCs）, i.e. , ASCs, （including plasma blasts, splenic plasma cells, and long-lived bone marrow plasma cells）. This signifcant change-over occurs to accommodate the massive amount of secretory-specific immunoglobulin that ASCs make and the export processes itself. It is well known that there is an up-regulation of a small number of ASC-specific transcription factors Prdm1 （B-lymphocyte-induced maturation protein 1）, interferon regulatory factor 4, and Xbp1, and the reciprocal down-regulation of Pax5, Bcl6 and Bach2, which maintain the B cell program. Less well appreciated are the major alterations in transcription elongation and RNA proce-ssing occurring between B cells and ASCs. The three ELL family members ELL1, 2 and 3 have different protein sequences and potentially distinct cellular roles in transcription elongation. ELL1 is involved in DNA repair and small RNAs while ELL3 was previously described as either testis or stem-cell specifc. After B cell stimulation to ASCs, ELL3 levels fall precipitously while ELL1 falls off slightly. ELL2 is induced at least 10-fold in ASCs relative to B cells. All of these changes cause the RNA Polymerase Ⅱ in ASCs to acquire different properties, leading to differences in RNA processing and histone modifcations.
文摘Background:Traumatic brain edema(TBE)is caused by a specific water channel mediated by membrane aquaporins.Aquaporin-4(AQP4)plays an especially important role in this process,but the relationship between AQP4 and TBE remains unclear.The purpose of this study was to explore expression of AQP4 in the hippocampus after traumatic brain injury(TBI),as well as the effect of brain edema on skeletal protein and its function in hippocampal neurons.Methods:The adult male Wistar rats we divided into a sham group and a TBI group,the latter of which was further divided into 1,3,6,12,24 and 72 hours(h)and 15 days(d)post injury subgroups.A proper TBI model was established,and brain edema was assessed in each group by water content.We measured the abundance of various proteins,including hypoxia inducible factor-1α(HIF-1α),AQP4,microtubule-associated protein 2(MAP2),tau-5 protein,phosphorylated level of TAU,synaptophysin,cyclic adenosine monophosphate response element binding protein(CREB),phosphorylated CREB and general control nonrepressed 2,in each group.Hippocampal neurons and spatial memory test were analyzed in different time points.Results:Compared with that in the sham group,the level of AQP4 in hippocampal neurons began to significantly increase at 1 h post TBI and then decreased at 15 d post TBI.During this time frame,AQP4 level peaked at 12 and 72 h,and these peaks were closely correlated with high brain water content.HIF-1αdisplayed a similar trend.Conversely,levels of MAP2 began to decrease at 1 h post TBI and then increase at 15 d post TBI.In addition,the most severe brain edema in rats was found at 24 h post TBI,with neuronal loss and hippocampal dendritic spine injury.Compared to those in the sham group,rats in the TBI groups had significantly prolonged latency and significantly shortened exploration time.Conclusions:AQP4 level was closely correlated with severity of brain edema,and abnormal levels thereof aggravated such severity after TBI.
