Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Effect of MCE on CCl 4 -induced acute liver damage in mice.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Photomicrographs of liver sections taken from mice treated with CCl 4 with or without pretreatment with MCE. (a) Normal; (b) CCl 4 ; (c) CCl 4 + MCE L ; (d) CCl 4 + MCE M ; and (e) CCl 4 + MCE H .

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Protective effect of MCE on the ultrastructure of hepatocytes induced by CCl 4 . Mice were divided into five groups: (a) Normal, (b) CCl 4 , (c) CCl 4 + MCE L , (d) CCl 4 + MCE M and (e) CCl 4 + MCE H . The CCl 4 and different MCE groups were administered saline or various concentrations of MCE orally for 5 days before the intra-peritoneal injection of 0.30% CCl 4 . Specimens were taken 24 h later and regularly prepared for examination under an electron microscope (×5000).

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques: Saline, Injection, Microscopy

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Prevention of MCE on mitochondrial membrane potential dissipation induced by CCl 4 . Mice were treated with MCE for 5 days before pretreatment with CCl 4 . Liver mitochondria were isolated and the mitochondrial membrane potential was determined using Rh123. Each value represents mean ± SD ( n = 8). * P < .05, ** P < .01, versus Normal; + P < .05 versus CCl 4 group.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques: Membrane, Isolation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Effect of MCE on liver mitochondrial calcium content in mice treated with CCl 4 . Mice were treated with MCE for 5 days before pretreatment with CCl 4 . Liver mitochondria were then isolated and mitochondrial free calcium content was determined using Fluo-3. MCE showed a dose-dependent suppression of the CCl 4 -induced intra-mitochondrial Ca 2+ overload. Values represent mean ± SD ( n = 6). * P < .01 versus the normal group; + P < .01 versus the CCl 4 group.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques: Isolation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Effect of MCE on Ca 2+ -induced mitochondrial swelling. CCl 4 can decrease the sensitivity in Ca 2+ -induced mitochondrial swelling. MCE dose-dependently recuperated this sensitivity. The curves represent typical recordings from experiments of at least three different mitochondrial preparations.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: Effect of MCE on mitochondrial VDAC expression in CCl 4 -insulted mouse livers. Livers from various groups were taken 24 h following 0.30% CCl 4 . (a) Inhibitory effect of MCE on the decrease in VDAC mRNA level induced by CCl 4 analyzed by RT-PCR. (b) Inhibitory effect of MCE on the decrease in VDAC protein level induced by CCl 4 analyzed by western blot.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Myrica rubra Extracts Protect the Liver from CCl 4 -Induced Damage

doi: 10.1093/ecam/nep196

Figure Lengend Snippet: CCl 4 could induce peroxidation of the unsaturated fatty acids of cell membrane, and lead to membrane injury, enhance the leakage of AST, the dissipation of mitochondrial membrane potential, hepatocellular Ca 2+ overload and decrease the sensitivity in Ca 2+ -induced mitochondrial swelling and other the mRNA and the protein of VDAC's expressions. Using MCE could prevent all these changes, that is to say, MCE has hepatoprotective activity and the mechanisms underlying its protective effects may be related to the mitochondrial protection.

Article Snippet: Et Zucc. extract (MCE) against liver injury induced by CCl 4 , addressing the possible action of MCE on liver mitochondrial and VDAC expression in order to search for the mechanism underlying its hepatoprotective activity.

Techniques: Membrane, Activity Assay

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Mixed neuronal-glial cerebrocortical cultures from WT or IFNAR1KO mice were incubated with IFNβ in increasing doses (from 500 to 5,000 U/ml) or BSA/PBS vehicle control for 0, 3, 6, 12 and 24 h. Total RNA was extracted from cell lysates, analyzed by qRT-PCR and normalized to GAPDH expression levels. ( a–d ) RNA expression is shown as fold change (FC) in relation to vehicle treated controls which were defined as baseline activity. ( e–h ) Time course for protein expression measured in cell-free supernatants for CCL3, CCL4, CCL5 and CXCL10 using a commercially available multiplex assay as described in Methods. Baseline protein expression in vehicle treated cell cultures is represented as 0 h time point. Values are mean ± s.e.m.; n = 3–5 independent experiments per ISG; ***p < 0.001, **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test. For clarity, the significance is only indicated for differences between treatments and baseline within each experimental group.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Incubation, Control, Quantitative RT-PCR, Expressing, RNA Expression, Activity Assay, Multiplex Assay

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: ( a ) Mixed neuronal-glial cerebrocortical cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL (200 pM) and mouse IFNβ (5,000 U/ml) in the presence and absence of neutralizing antibodies against CCL3, CCL4, CCL5, IFNγ or CXCL10. IFNγ antibody was used as control for neutralizing antibodies since this protein was undetectable in cerebrocortical cell cultures. ( b ) Mouse cerebrocortical cultures from IFNAR1KO mice were stimulated with gp120 BaL for 24 h the presence or absence of mouse IFNβ (5,000 U/ml) or BSA/PBS control. ( c ) Cerebrocortical cell cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL in the presence and absence of murine CCL4 (2 or 20 nM). Neuronal survival was assessed by immunofluorescence microscopy and counting of MAP-2/NeuN double-positive neurons. Values are mean ± s.e.m.; n = 3–5 independent experiments with 3–7 replicates and an average of 9,000 (IFNAR1KO) or 5,700 (WT) cells counted per condition; ** p < 0.01, *** p < 0.001 by ANOVA with Fisher’s PLSD post hoc test.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Control, Immunofluorescence, Microscopy

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: RNA was purified from one brain hemisphere each of 4–5 month-old HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle and analyzed by qRT-PCR for fold-change (FC) in ISG expression. Significant changes in gene expression were observed between IFNβ and vehicle treatment groups in WT brains ( a ) for CCL4, and in gp120tg brains ( b ) for CCL4, CXCL11 and IRF3. Expression of transgenic HIVgp120 was not affected by IFNβ ( c ). Values are mean ± s.e.m.; n = 4–5 animals per group/genotype; *p < 0.05, student’s t-test.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Purification, Quantitative RT-PCR, Expressing, Gene Expression, Transgenic Assay

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Sagittal brains sections of HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle (veh) were immunolabeled for CCL4, neuronal MAP-2 or astrocytic GFAP. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was labeled with Hoechst (H) 33342. The fluorescence-labeled brain sections were analyzed using confocal laser-scanning microscopy. Representative images of cortex layer III are shown; scale bar, 50 μm.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Immunolabeling, Labeling, Fluorescence, Confocal Laser Scanning Microscopy

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: ( a ) Cerebrocortical cultures from mice were prepared to either contain microglia, neurons and astrocytes (M + N + A) or were depleted of microglia (N + A) or neurons and microglia (A). Complete and depleted cell cultures were incubated with mIFNβ (5,000 U/ml) or BSA/PBS vehicle control for 0, 3, 6, 12 and 24 h and concentrations of CCL4 were measured in cell-free supernatants using a commercially available multiplex assay as described in Methods. Maximum concentrations were reached in samples of 12 to 24 h mIFNβ exposure and compared to vehicle-treated, baseline samples. Values are mean ± s.e.m.; n = 3 independent experiments; *p < 0.05, student’s t-test. ( b ) Microglia-depleted rat cerebrocortical cultures were exposed for 24 h to 50% cell-free conditioned media (CM) from human MDM in the presence or absence of human IFNβ (5,000 U/ml). MDM were previously stimulated for 24 h with HIV-1 gp120 BaL (MDM gp120 CM) or vehicle (MDM CM). Following the incubation the cells were fixed and permeabilized. Neurons were immunolabeled for neuronal MAP-2 and NeuN and nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments, with 4–8 replicates and an average of 4,000 cells counted per condition; **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Incubation, Control, Multiplex Assay, Immunolabeling, Staining, Fluorescence, Microscopy, Cell Counting