Objective To explore the role of HIV-1 tat gene variations in AIDS dementia complex （ADC） pathogenesis. Methods HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC an...Objective To explore the role of HIV-1 tat gene variations in AIDS dementia complex （ADC） pathogenesis. Methods HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA. Results HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area （38-47aa）. All Tat proteins could induce ug7 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen. The level of Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen were obviously higher than that from the non-ADC patient＇s basal ganglia. Conclusion Tat protein core functional area （38-47aa） may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.展开更多
It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of...It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of vascular dementia. In model rats, hippocampal CA1 neurons were severely damaged, and expression of the autophagy-related proteins beclin-1, cathepsin B and microtubule-associated protein 1 light chain 3 was elevated compared with that in sham-operated animals. These responses were suppressed in animals that received a single intraperitoneal injection of wortmannin, an autophagy inhibitor, prior to model establishment. The present results confirm that autophagy and autophagy-related proteins are involved in the pathological changes of vascular dementia, and that inhibition of autophagy has neuroprotective effects.展开更多
Highly active antiretroviral treatment(HAART) has had a significant impact on survival of individuals with acquired immunodeficiency syndrome(AIDS);however,with the longer life-span of patients with AIDS,there is incr...Highly active antiretroviral treatment(HAART) has had a significant impact on survival of individuals with acquired immunodeficiency syndrome(AIDS);however,with the longer life-span of patients with AIDS,there is increasing prevalence of AIDS dementia complex(ADC) and other non-AIDS-defining illness,and cardiovascular diseases(CVD) are also common.The influence of these varied disease processes on HIV-1 DNA concentration in brain tissues has not been thoroughly assessed in the post-HAART era.The purpose of the current study is to clarify the impacts of ADC and other complications of HIV disease on the viral load in the brains in AIDS patients with post-HARRT.We examined autopsy specimens from the brains of thirteen patients who died from complications of AIDS with quantitative polymerase chain reaction(QPCR).All but one patient had received HAART prior to death since 1995.Two patients died with severe CVD,multiple cerebrovascular atherosclerosis(CVA) throughout the brain and five patients died with ADC.Six patients had no ADC/CVA.A QPCR was used to measure the presence of HIV-1 DNA in six brain tissues(meninges,frontal grey matter,frontal white matter,temporal subcortex,cerebellum and basal ganglia).In the post-HARRT era,for non-ADC/CVA patients,HIV-1 DNA concentration in brain tissues was statistically higher than that in patients with ADC.In a new finding,two patients who suffered from severe CVD,especially CVA,also had high concentrations of HIV-1 in brain compartments not showing ADC related changes.To our knowledge,this is the first report of a relationship between the CVA and HIV-1 viral burden in brain.The current observations suggest that HAART-resistant HIV reservoirs may survive within ADC lesions of the brain as well as the macrophage rich atherosclerosis,which needs to be confirmed by more AIDS cases with CVA.展开更多
Background HIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-a （TNF-a） and interleukin-113 （IL-113）, which play major role in the neuronal death. It h...Background HIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-a （TNF-a） and interleukin-113 （IL-113）, which play major role in the neuronal death. It has been shown that different HIV-1 variants have varying abilities to elicit secretion of TNF-a by peripheral blood mononuclear cell （PBMC）; however, whether the difference of gp120 gene could affect the secretion of TNF-a and IL-113 by glial cells is unknown. The aim of this study was to explore the association between gene diversity and induction of neurotoxic cytokines. Methods In this study, we constructed retroviral vectors MSCV-IRES-GFP/gp120 using HIV-1 gp120 genes isolated from four different tissues of one patient who died of AIDS dementia complex （ADC）. Recombinant retroviruses produced by cotransfection of MSCV-IRES-GFP/gp120, pCMV-VSV-G and pUMVC into 293T cells were collected and added into U87 glial cells. Concentrations of TNF-α and IL-1β secreted by transduced U87 cells were assayed with ELISA separately. Results The four HIV-1 gp120 were in the different branch of the neighbor-joining tree. Compared to the pMIG retrovirus （gp120-negative） or U87 cells, all the gp120-positive recombinant retroviruses induced more TNF-a （P 〈0.01） and IL-113 （P 〈0.01）. In addition, compared with the L/MIG retrovirus, all the three brain gp120-positive recombinant retroviruses induced less TNF-α （P 〈0.01） and IL-1β （P 〈0.01）. Conclusions HIV-1 gp120 could induce U87 cells secret more TNF-α and IL-1β again. The more important is that difference of HIV-1 gp120, especially cell-tropism may account for the different ability in eliciting secretion of TNF-α and IL-1 β, which might supply a novel idea helping understand the pathogenesis of ADC.展开更多
BACKGROUND： Previous studies of curcumin have focused mainly on its cytotoxic properties for antitumor therapy. There are few studies addressing the application of curcumin in the prevention and treatment of nervous ...BACKGROUND： Previous studies of curcumin have focused mainly on its cytotoxic properties for antitumor therapy. There are few studies addressing the application of curcumin in the prevention and treatment of nervous system diseases. OBJECTIVE： To observe the protective effect of curcumin against tumor necrosis factor-alpha （TNF-α）-induced neuronal damage in the rat hippocampus and to explore the intervention effect of curcumin on Ca^2＋ influx following neuronal damage. DESIGN, TIME AND SETTING： A cell morphological and physiological study was performed at the Institute of Brain Research, Medical College of Jinan University, China, from December 2006 to June 2007. MATERIALS： Curcumin （Sigma, USA） and TNF-α （Sigma, USA） were used in this study. METHODS： Hippocampal neurons were isolated from one-day neonatal rats and primarily cultured for 5 days. Following this they received 1 pmol/L curcumin and 100 ng/mL TNF-a pre-treatment. Dynamic morphological changes were observed for 1 hour by inverted microscopy. At 48 hours post-treatment, static morphological characteristics of the neurons were observed using inverted microscopy. Subsequently, hippocampal neurons were primarily cultured for 7 days, after receiving 1 pmol/L curcumJn and 4.5 ng/mL TNF-a pre-treatment. Intracellular free Ca^2＋ was measured using Fluo 3/acetoxymethyl ester. MAIN OUTCOME MEASURES： Effects of curcumin on TNF-a-induced neuronal damage and Ca^2＋ influx in the rat hippocampus were measured. RESULTS： Following curcumin treatment, TNF-a-induced neurons grew as normal. TNF-a induced a rapid Ca^2＋ influx into the neuronal cytoplasm; however, Ca2＋ fluorescence intensity only slightly increased when neurons were co-perfused with curcumin and TNF-α. CONCLUSION： Curcumin has a protective effect on rat hippocampal neurons possibly by reducing the TNF-α-induced rapid Ca^2＋ influx into neuronal cytoplasm and by maintaining the Ca^2＋ homeostasis.展开更多
基金supported by the Science&Technology Development Program of Shandong Province(Grant No.2007GG30002003)
文摘Objective To explore the role of HIV-1 tat gene variations in AIDS dementia complex （ADC） pathogenesis. Methods HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA. Results HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area （38-47aa）. All Tat proteins could induce ug7 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen. The level of Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen were obviously higher than that from the non-ADC patient＇s basal ganglia. Conclusion Tat protein core functional area （38-47aa） may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.
基金supported by the Scientific Technology Research Project of Hebei Provincial Higher Learning Schools in China,No.ZH2012046the Major Medical Research Program of Hebei Province in China,No.ZD2013087
文摘It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of vascular dementia. In model rats, hippocampal CA1 neurons were severely damaged, and expression of the autophagy-related proteins beclin-1, cathepsin B and microtubule-associated protein 1 light chain 3 was elevated compared with that in sham-operated animals. These responses were suppressed in animals that received a single intraperitoneal injection of wortmannin, an autophagy inhibitor, prior to model establishment. The present results confirm that autophagy and autophagy-related proteins are involved in the pathological changes of vascular dementia, and that inhibition of autophagy has neuroprotective effects.
基金Supported by the National Institutes of Health (Grant Nos. NIH ZMH1 BRB-S and UOI CA66259-09 TDC)National Science Foundation (Grant No. NSF DMI0349669)abd Science & Technology Development Program of Shandong Province (Grant No. 2007GG30002003)
文摘Highly active antiretroviral treatment(HAART) has had a significant impact on survival of individuals with acquired immunodeficiency syndrome(AIDS);however,with the longer life-span of patients with AIDS,there is increasing prevalence of AIDS dementia complex(ADC) and other non-AIDS-defining illness,and cardiovascular diseases(CVD) are also common.The influence of these varied disease processes on HIV-1 DNA concentration in brain tissues has not been thoroughly assessed in the post-HAART era.The purpose of the current study is to clarify the impacts of ADC and other complications of HIV disease on the viral load in the brains in AIDS patients with post-HARRT.We examined autopsy specimens from the brains of thirteen patients who died from complications of AIDS with quantitative polymerase chain reaction(QPCR).All but one patient had received HAART prior to death since 1995.Two patients died with severe CVD,multiple cerebrovascular atherosclerosis(CVA) throughout the brain and five patients died with ADC.Six patients had no ADC/CVA.A QPCR was used to measure the presence of HIV-1 DNA in six brain tissues(meninges,frontal grey matter,frontal white matter,temporal subcortex,cerebellum and basal ganglia).In the post-HARRT era,for non-ADC/CVA patients,HIV-1 DNA concentration in brain tissues was statistically higher than that in patients with ADC.In a new finding,two patients who suffered from severe CVD,especially CVA,also had high concentrations of HIV-1 in brain compartments not showing ADC related changes.To our knowledge,this is the first report of a relationship between the CVA and HIV-1 viral burden in brain.The current observations suggest that HAART-resistant HIV reservoirs may survive within ADC lesions of the brain as well as the macrophage rich atherosclerosis,which needs to be confirmed by more AIDS cases with CVA.
文摘Background HIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-a （TNF-a） and interleukin-113 （IL-113）, which play major role in the neuronal death. It has been shown that different HIV-1 variants have varying abilities to elicit secretion of TNF-a by peripheral blood mononuclear cell （PBMC）; however, whether the difference of gp120 gene could affect the secretion of TNF-a and IL-113 by glial cells is unknown. The aim of this study was to explore the association between gene diversity and induction of neurotoxic cytokines. Methods In this study, we constructed retroviral vectors MSCV-IRES-GFP/gp120 using HIV-1 gp120 genes isolated from four different tissues of one patient who died of AIDS dementia complex （ADC）. Recombinant retroviruses produced by cotransfection of MSCV-IRES-GFP/gp120, pCMV-VSV-G and pUMVC into 293T cells were collected and added into U87 glial cells. Concentrations of TNF-α and IL-1β secreted by transduced U87 cells were assayed with ELISA separately. Results The four HIV-1 gp120 were in the different branch of the neighbor-joining tree. Compared to the pMIG retrovirus （gp120-negative） or U87 cells, all the gp120-positive recombinant retroviruses induced more TNF-a （P 〈0.01） and IL-113 （P 〈0.01）. In addition, compared with the L/MIG retrovirus, all the three brain gp120-positive recombinant retroviruses induced less TNF-α （P 〈0.01） and IL-1β （P 〈0.01）. Conclusions HIV-1 gp120 could induce U87 cells secret more TNF-α and IL-1β again. The more important is that difference of HIV-1 gp120, especially cell-tropism may account for the different ability in eliciting secretion of TNF-α and IL-1 β, which might supply a novel idea helping understand the pathogenesis of ADC.
基金the grant from Medical Science Foundation of Guangdong Province,No.A2006334the Natural Science Foundation of Guangdong Province,No.06105246,No.9151040701000008the Science and Technology Project of Guangzhou City,No.2007J1-C0041
文摘BACKGROUND： Previous studies of curcumin have focused mainly on its cytotoxic properties for antitumor therapy. There are few studies addressing the application of curcumin in the prevention and treatment of nervous system diseases. OBJECTIVE： To observe the protective effect of curcumin against tumor necrosis factor-alpha （TNF-α）-induced neuronal damage in the rat hippocampus and to explore the intervention effect of curcumin on Ca^2＋ influx following neuronal damage. DESIGN, TIME AND SETTING： A cell morphological and physiological study was performed at the Institute of Brain Research, Medical College of Jinan University, China, from December 2006 to June 2007. MATERIALS： Curcumin （Sigma, USA） and TNF-α （Sigma, USA） were used in this study. METHODS： Hippocampal neurons were isolated from one-day neonatal rats and primarily cultured for 5 days. Following this they received 1 pmol/L curcumin and 100 ng/mL TNF-a pre-treatment. Dynamic morphological changes were observed for 1 hour by inverted microscopy. At 48 hours post-treatment, static morphological characteristics of the neurons were observed using inverted microscopy. Subsequently, hippocampal neurons were primarily cultured for 7 days, after receiving 1 pmol/L curcumJn and 4.5 ng/mL TNF-a pre-treatment. Intracellular free Ca^2＋ was measured using Fluo 3/acetoxymethyl ester. MAIN OUTCOME MEASURES： Effects of curcumin on TNF-a-induced neuronal damage and Ca^2＋ influx in the rat hippocampus were measured. RESULTS： Following curcumin treatment, TNF-a-induced neurons grew as normal. TNF-a induced a rapid Ca^2＋ influx into the neuronal cytoplasm; however, Ca2＋ fluorescence intensity only slightly increased when neurons were co-perfused with curcumin and TNF-α. CONCLUSION： Curcumin has a protective effect on rat hippocampal neurons possibly by reducing the TNF-α-induced rapid Ca^2＋ influx into neuronal cytoplasm and by maintaining the Ca^2＋ homeostasis.
