Journal: Frontiers in Cell and Developmental Biology

Article Title: Detecting Key Functional Components Group and Speculating the Potential Mechanism of Xiao-Xu-Ming Decoction in Treating Stroke

doi: 10.3389/fcell.2022.753425

Figure Lengend Snippet: The comparation of effective components and non-KFCG components on cell viabilities induced PC12 cells. (A–E) The treatments with ferulic acid, vanillic acid, zingerone, caryophyllene oxide, and methylephedrine hydrochloride, respectively. (F) The structures of ferulic acid, vanillic acid, zingerone, caryophyllene oxide, methylephedrine hydrochloride, and edaravone. K represents the control group without the treatments of components and OGD; M represents the model group with OGD treatment and without components treatments; Eda represents edaravone (20 μM). ### represents the comparation of model group to control group ( p = 0.001); *, **, and *** represent the comparations of components groups to model group in the levels of p < 0.05, p < 0.01, and p < 0.001. (G) The screening process of components. Five components marked in red were selected for the in vitro experiments.

Article Snippet: The PC12 cell line was obtained from CHI SCIENTIFIC (Shanghai, China) and cultured in RPMI-1640 with FBS, penicillin 100 U/mL, streptomycin 100 μg/ml, respectively, at 37°C in a fully humidified 5% CO 2 atmosphere.

Techniques: Control, In Vitro

Journal: Cell Death & Disease

Article Title: MiR-127-3p targeting CISD1 regulates autophagy in hypoxic–ischemic cortex

doi: 10.1038/s41419-021-03541-x

Figure Lengend Snippet: A Results of miRNA sequencing analysis showed that the expression of miRNAs in sham and HI have obvious difference ( n = 2/group). B The qRT-PCR validation of differential miRNAs in normal group rats and HI group rats at 12 h after HI injury. Each bar was the mean ± s.d. relative to −log2 (** P < 0.01 with Student’s t -test, n = 6). C Relative expression of miRNA-127-3p in heart, liver, brain, and kidney of rats. MiRNA-127-3p was selectively highly expressed in the brain (** P < 0.01 with one-way ANOVA, n = 6). D qRT-PCR analysis of miR-127-3p expression at 12, 48, and 96 h. MiRNA-127-3p decreased at 12 h after HI, while it increased at 48 and 96 h after HI injury (** P < 0.01 with one-way ANOVA, n = 6). E The relative expression of miR-127-3p in OGD of PC12, SY5Y, and neurons was significantly decreased in vitro (** P < 0.01 with Student’s t -test, n = 6). HI hypoxic ischemic.

Article Snippet: PC12 cell line (GCC-KI0009RT) was obtained from Shanghai Genechem Co., LTD and SY5Y cell line (BFN60700126) was obtained from BLUEFBIO.

Techniques: Sequencing, Expressing, Quantitative RT-PCR, Biomarker Discovery, In Vitro

Journal: Basic and Clinical Neuroscience

Article Title: Effectiveness of Nicotinamide Phosphoribosyltransferase/Pre-B Cell Colony-enhancing Factor/Visfatin in preventing High Glucose-induced Neurotoxicity in an In-vitro Model of Diabetic Neuropathy

doi: 10.32598/bcn.2021.2870.2

Figure Lengend Snippet: A) The effects of different concentrations of glucose on pheochromocytoma PC12 cell viability, B) The effects of visfatin on high-glucose damaged PC12 cells Data are shown as Mean±SEM; * P<0.05, ** P<0.01 vs control, # P<0.001, ## P<0.0001 vs HG treatment.

Article Snippet: PC12 cells were purchased from the Buali (Avicenna) Research Institute.

Techniques: Control

Journal: Basic and Clinical Neuroscience

Article Title: Effectiveness of Nicotinamide Phosphoribosyltransferase/Pre-B Cell Colony-enhancing Factor/Visfatin in preventing High Glucose-induced Neurotoxicity in an In-vitro Model of Diabetic Neuropathy

doi: 10.32598/bcn.2021.2870.2

Figure Lengend Snippet: A) The effect of visfatin on HG-induced intracellular ROS level in PC12 cells, B) The effects of visfatin on HG-induced apoptosis of PC12 cell, C) The apoptosis evaluation of PC12 cells under different treatments Note: Data are shown as Mean±SEM; * P<0.05 vs control, # P<0.01, ## P<0.0001vs HG treatment.

Article Snippet: PC12 cells were purchased from the Buali (Avicenna) Research Institute.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology-Guided Discovery of Traditional Chinese Medicine Extracts for Alzheimer’s Disease: Targeting Neuroinflammation and Gut–Brain Axis Dysfunction

doi: 10.3390/ijms26178545

Figure Lengend Snippet: Cell viability assessment of PC12/Caco-2 co-cultures treated with TCM extracts and active compounds. ( A ) Cell viability of PC12/Caco-2 co-cultures exposed to Aβ and LPS damage, followed by treatment with 20 μg/mL concentrations of VT, PM, AVF, and EF extracts. ( B ) Cell viability of PC12/Caco-2 co-cultures exposed to Aβ and LPS damage, followed by treatment with 2 μM concentrations of bioactive compounds. PC12 cells were initially stimulated with NGF to enhance neuronal differentiation (1.5–2-fold increase in proliferation). Co-cultured cells were then pre-treated with 10 μM Aβ or LPS for 1 h to induce cytotoxicity. Subsequently, PC12 cells (basal layer) were treated with herbal extracts (10, 20, 50 μg/mL) or bioactive compounds (2, 5 μM) in low NGF medium, while Caco-2 cells (upper layer) received the same concentrations of LPS treatment. Donepezil hydrochloride served as positive control. After 24 h treatment, cell viability was assessed using the MTT assay. Data are presented as mean values. Different superscript letters (a, b, c) indicate significant differences between groups within each parameter (Tukey’s test, p < 0.05). Abbreviations: Aβ, β-amyloid; AVF, Apocyni Veneti Folium ; EF, Eucommiae folium ; LPS, lipopolysaccharide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NGF, nerve growth factor; PM, Plantago major ; TCM, traditional Chinese medicine; VT, Vitex trifolia .

Article Snippet: An in vitro gut–brain axis model was established using a co-culture system of PC12 neuronal cells and Caco-2 intestinal epithelial cells (KCLB 21721, Korean Cell Line Bank, Seoul, Republic of Korea), as illustrated in .

Techniques: Cell Culture, Positive Control, MTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology-Guided Discovery of Traditional Chinese Medicine Extracts for Alzheimer’s Disease: Targeting Neuroinflammation and Gut–Brain Axis Dysfunction

doi: 10.3390/ijms26178545

Figure Lengend Snippet: Relative mRNA expression of PC12 cells/Caco-2 cells co-culture. ( A ) Inflammatory cytokine mRNA expression ( TNF-α , IL-1β , IL-6 ) in PC12 and Caco-2 cells following treatment with 20 μg/mL concentrations of VT, PM, AVF, and EF extracts. ( B ) Neurodegeneration-related mRNA expression ( Tau and BDNF ) in PC12 cells following treatment with 20 μg/mL concentrations of VT, PM, AVF, and EF extracts. ( C ) Inflammatory cytokine mRNA expression ( TNF-α , IL-1β , IL-6 ) in PC12 and Caco-2 cells following treatment with 2 μM concentrations of active ingredients. ( D ) Neurodegeneration-related mRNA expression ( Tau and BDNF ) in PC12 cells following treatment with 2 μM concentrations of active ingredients. PC12 and Caco-2 cells were maintained in co-culture. PC12 cells were exposed to 10 μM Aβ for 1 h to induce neurodegeneration, while Caco-2 cells were exposed to 10 μM LPS for 1 h to induce intestinal inflammation. Following damage induction, cells were treated for 24 h with vehicle control (cell water), donepezil hydrochloride (positive control), TCM extracts (VT, PM, AVF, EF), or bioactive compounds. mRNA was then extracted from both cell types for gene expression analysis. Data are presented as mean ± standard error. Different letters (a, b, c, d) above bars indicate significant differences between treatment groups (Tukey’s test, p < 0.05). Identical letters indicate no significant difference. Abbreviations: Aβ, β-amyloid; AVF, Apocyni Veneti Folium ; BDNF, brain-derived neurotrophic factor; EF, Eucommiae folium ; IL-1β, interleukin-1β; IL-6, interleukin-6; LPS, lipopolysaccharide; PM, Plantago major ; TCM, traditional Chinese medicine; TNF-α, tumor necrosis factor-α; VT, Vitex trifolia .

Article Snippet: An in vitro gut–brain axis model was established using a co-culture system of PC12 neuronal cells and Caco-2 intestinal epithelial cells (KCLB 21721, Korean Cell Line Bank, Seoul, Republic of Korea), as illustrated in .

Techniques: Expressing, Co-Culture Assay, Control, Positive Control, Gene Expression, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology-Guided Discovery of Traditional Chinese Medicine Extracts for Alzheimer’s Disease: Targeting Neuroinflammation and Gut–Brain Axis Dysfunction

doi: 10.3390/ijms26178545

Figure Lengend Snippet: Insulin signaling pathway in PC12 cells by Western blot. ( A ) Representative Western blot images showing protein expression levels of key insulin signaling pathway components (Akt, GSK-3β, AMPK) and their phosphorylated forms in PC12 cells from co-culture experiments. ( B ) Quantitative analysis of total and phosphorylated protein levels of Akt, GSK-3β, and AMPK in PC12 cells following treatment with TCM extracts. Experimental protocol: PC12 and Caco-2 cells were maintained in co-culture with PC12 cells in the basal layer and Caco-2 cells in the upper layer. PC12 cells were exposed to 10 μM Aβ for 1 h to induce neuronal damage, while Caco-2 cells were simultaneously exposed to 10 μM LPS for 1 h to simulate intestinal inflammation. Following damage induction, cells were treated for 24 h with vehicle control, donepezil hydrochloride (positive control), or 20 μg/mL concentrations of VT, PM, AVF, and EF extracts. PC12 cells were then harvested for Western blot analysis of insulin signaling pathway proteins, including total and phosphorylated forms of Akt, AMPK, and GSK-3β, with β-actin serving as loading control. Data are presented as mean ± standard error. Different letters (a, b, c) above bars indicate significant differences between treatment groups (Tukey’s test, p < 0.05). Identical letters indicate no significant difference. Abbreviations: Akt, AKT serine/threonine kinase; AMPK, AMP-activated protein kinase; AVF, Apocyni Veneti Folium ; EF, Eucommiae folium ; GSK-3β, glycogen synthase kinase-3β; LPS, lipopolysaccharide; PM, Plantago major ; TCM, traditional Chinese medicine; VT, Vitex trifolia ; Aβ, β-amyloid.

Article Snippet: An in vitro gut–brain axis model was established using a co-culture system of PC12 neuronal cells and Caco-2 intestinal epithelial cells (KCLB 21721, Korean Cell Line Bank, Seoul, Republic of Korea), as illustrated in .

Techniques: Western Blot, Expressing, Co-Culture Assay, Control, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Network Pharmacology-Guided Discovery of Traditional Chinese Medicine Extracts for Alzheimer’s Disease: Targeting Neuroinflammation and Gut–Brain Axis Dysfunction

doi: 10.3390/ijms26178545

Figure Lengend Snippet: Morphological and mitochondrial analysis of PC12/Caco-2 co-cultures. ( A ) Hematoxylin and eosin (H&E) staining of Caco-2 cells following treatment with 20 μg/mL TCM extracts in co-culture experiments (100× magnification). ( B ) Toluidine blue staining of Caco-2 cells showing mast cell infiltration (indicated by red circles) following treatment with 20 μg/mL TCM extracts in co-culture experiments (100× magnification). ( C ) JC-1 fluorescent staining of PC12/Caco-2 co-cultures demonstrating mitochondrial membrane potential and cellular protection following treatment with TCM extracts (20 μg/mL) or active ingredients (2 μM) (200× magnification). Experimental protocol: PC12 and Caco-2 cells were maintained in co-culture with PC12 cells in the basal layer and Caco-2 cells in the upper layer. PC12 cells were exposed to 10 μM Aβ for 1 h, while Caco-2 cells were exposed to 10 μM LPS for 1 h to induce cellular damage. Following damage induction, cells were treated for 24 h with vehicle control, donepezil hydrochloride (positive control), VT, PM, AVF, or EF extracts (20 μg/mL), or active ingredients (2 μM). JC-1 fluorescent staining was used to examine the degree of apoptosis in Caco-2 cells and PC12 cells. The mitochondrial membrane potential was expressed as red fluorescence. The more fluorescent red (Caco-2 cells) and fluorescent green (PC12-cells), the higher the intensity of cell protection and the inhibition of cell apoptosis. Data are presented as mean ± standard error. Different letters (a, b, c) above bars indicate significant differences between treatment groups (Tukey’s test, p < 0.05). Identical letters indicate no significant difference. Abbreviations: AVF, Apocyni Veneti Folium; EF, Eucommiae folium; H&E, hematoxylin and eosin; JC-1, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; LPS, lipopolysaccharide; PM, Plantago major ; VT, Vitex trifolia .

Article Snippet: An in vitro gut–brain axis model was established using a co-culture system of PC12 neuronal cells and Caco-2 intestinal epithelial cells (KCLB 21721, Korean Cell Line Bank, Seoul, Republic of Korea), as illustrated in .

Techniques: Staining, Co-Culture Assay, Membrane, Control, Positive Control, Fluorescence, Inhibition