Journal: International Journal of Molecular Sciences

Article Title: TOMM40 Genetic Variants Cause Neuroinflammation in Alzheimer’s Disease

doi: 10.3390/ijms24044085

Figure Lengend Snippet: TOMM40 genetic variants induce the secretion of pro-inflammatory cytokines in microglial cells, leading to cell death of hippocampal neurons. ( A ) Compared to control cells or cells expressing WT TOMM40, BV2 microglial cells’ expression of (F113L) or (F131L) TOMM40 significantly increased secretion of pro-inflammatory IL-1β, IL-6, or TNF-α in culture medium of BV2 microglial cells. ( B ) Culture medium (CM) of HT22 hippocampal neurons was replaced with CM from BV2 microglial cells transfected with cDNA of WT, (F113L) or (F131L) TOMM40. One day after replacement, CM of BV2 microglia cells expressing mutant (F113L) or (F131L) TOMM40 significantly reduced cell viability of HT22 hippocampal neurons. Each bar represents mean ± S.D. of four experiments. Each experiment was performed in triplicate. * p < 0.05 or ** p < 0.01 compared to control BV2 microglial cells or HT22 hippocampal neurons.

Article Snippet: BV2 mouse microglial cells and HT22 mouse hippocampal neuronal cells were purchased from Elabscience (Cat. EP-ML-0697 and EP-CL-0493) and maintained in a DMEM medium containing 10% FBS.

Techniques: Control, Expressing, Transfection, Mutagenesis

Journal: Antioxidants

Article Title: Tetramethylpyrazine Analogue T-006 Protects Neuronal and Endothelial Cells Against Oxidative Stress via PI3K/AKT/mTOR and Nrf2 Signaling

doi: 10.3390/antiox13101272

Figure Lengend Snippet: Effects of T-006 in HT22 cells challenged by excessive glutamate. ( A ) Chemical structure of TMP and T-006. ( B ) Cell viability of HT22 cells treated with different concentrations of glutamate as measured by MTT assay. ( C ) Representative images of HT22 cells morphology under glutamate challenge (scale bar, 50 μm). ( D ) Schematic diagram of experiment protocol. HT22 cells were incubated with T-006 (1, 3, and 10 μM) or TMP (10 μM) for 24 h in the presence of glutamate. ( E ) Cell viability of HT22 cells treated with different concentrations of T-006 and TMP in the presence of glutamate (10 mM) as measured by MTT assay. Data are shown as mean ± SEM from at least three independent experiments (n > 3) and analyzed by one-way ANOVA followed by Tukey test. ### p < 0.001 vs. control group, *** p < 0.001 vs. glutamate-induced group.

Article Snippet: Mouse hippocampal neuronal cell line HT22 was generously provided by Prof. Yonghua Zhao (Institute of Chinese Medical Sciences, University of Macau).

Techniques: MTT Assay, Incubation, Control

Journal: Antioxidants

Article Title: Tetramethylpyrazine Analogue T-006 Protects Neuronal and Endothelial Cells Against Oxidative Stress via PI3K/AKT/mTOR and Nrf2 Signaling

doi: 10.3390/antiox13101272

Figure Lengend Snippet: T-006 inhibited glutamate-induced ROS production and apoptosis in HT22 cells. ( A , B ) Quantitative analysis of ROS generation by flow cytometry with H2DCFHDA. ( C , D ) Quantitative analysis of apoptosis by flow cytometry with Annexin V-FITC/PI. Data are shown as mean ± SEM from at least three independent experiments (n > 3) and analyzed by one-way ANOVA followed by Tukey test. ### p < 0.001 vs. control group, *** p < 0.001 vs. glutamate-induced group. “ns” stands for “not significant”.

Article Snippet: Mouse hippocampal neuronal cell line HT22 was generously provided by Prof. Yonghua Zhao (Institute of Chinese Medical Sciences, University of Macau).

Techniques: Flow Cytometry, Control

Journal: Antioxidants

Article Title: Tetramethylpyrazine Analogue T-006 Protects Neuronal and Endothelial Cells Against Oxidative Stress via PI3K/AKT/mTOR and Nrf2 Signaling

doi: 10.3390/antiox13101272

Figure Lengend Snippet: T-006 induced protective effect via AKT-mediated mTOR and HO-1 signaling in glutamate-challenged HT22 cells. ( A ) Representative immunoblots of AKT, mTOR, LC3, and p62 proteins. ( B – E ) Quantitative analysis for p-mTOR, p-AKT, LC3-I/II, and p62 proteins levels (n = 3–4). ( F ) Representative immunoblots of Nrf2 and HO-1 proteins. ( G , H ) Quantitative analysis for Nrf2 and HO-1 (n = 3–4). Data are shown as mean ± SEM and analyzed by one-way ANOVA followed by Tukey test. # p < 0.05 and ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. glutamate-induced group.

Article Snippet: Mouse hippocampal neuronal cell line HT22 was generously provided by Prof. Yonghua Zhao (Institute of Chinese Medical Sciences, University of Macau).

Techniques: Western Blot, Control

Journal: Antioxidants

Article Title: Tetramethylpyrazine Analogue T-006 Protects Neuronal and Endothelial Cells Against Oxidative Stress via PI3K/AKT/mTOR and Nrf2 Signaling

doi: 10.3390/antiox13101272

Figure Lengend Snippet: T-006 induced protective effects in glutamate-challenged HT22 cells was inhibited by LY294002. ( A ) Representative images of cell morphology with bright field microscope (scale bar 100 μm). ( B ) Cell viability was detected by MTT assay (n = 6). ( C ) Representative immunoblots of AKT and mTOR proteins. ( D , E ) Quantitative analysis for p-mTOR and p-AKT (n = 3–4). ( F ) Representative immunoblots of Nrf2 and HO-1 proteins. ( G , H ) Quantitative analysis for Nrf2 and HO-1 (n = 3–4). Data are shown as mean ± SEM and analyzed by one-way ANOVA followed by Tukey test. # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. control group; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. glutamate-induced group. “ns” stands for “not significant”.

Article Snippet: Mouse hippocampal neuronal cell line HT22 was generously provided by Prof. Yonghua Zhao (Institute of Chinese Medical Sciences, University of Macau).

Techniques: Microscopy, MTT Assay, Western Blot, Control