hgmcs  (ScienCell)

Journal: Food Science of Animal Resources

Article Title: Oral Administration of Lactilactobacillus curvatus LB-P9 Promotes Hair Regeneration in Mice

doi: 10.5851/kosfa.2023.e74

Figure Lengend Snippet: (A) Both hFDPCs (1×10 4 cells/well) and hGMCs (1×10 4 cells/well) were cultured with 0.25% or 0.5% of LB-P9 CM at 96-well plate for 24 h. After culture, the relative proliferation ratio was measured using colorimetric assays (n=3). Vehicle, PBS treatment. Percentage indicated the concentration of LB-P9 CM in culture media. (B) The hGMCs (2×10 5 cells/well) were cultured with LB-P9 (2×10 6 CFU/well) at 6-well plate for 24 h. After culture, qRT-PCR analyses were performed to determine the relative expression levels of Vegf and Fgf7 in hGMCs (n=3). Vehicle, PBS treatment; LB-P9, LB-P9 treatment. All results are shown as the mean±SD. Significant differences compared with the control group are indicated as “p=number”. hFDPC, hair follicle dermal papilla cell; hGMC, hair germinal matrix cell; VEGF, vascular endothelial growth factor; FGF-7, fibroblast growth factor 7; CM, conditioned medium; PBS, phosphate-buffered saline; CFU, colony forming unit; qRT-PCR, quantitative reverse-transcriptase real-time polymerase chain reaction.

Article Snippet: Human hair germinal matrix cells (hGMCs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Cell Culture, Concentration Assay, Quantitative RT-PCR, Expressing, Control, Saline, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: The expression of ZFAS1 in DN blood samples and HG-induced HGMC ( A and B ) ZFAS1 is highly upregulated DN samples and HG-induced HGMC compared to the normal healthy or control group

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: The lncRNA ZFAS1 regulates HG-induced proliferation, oxidative stress, fibrosis, and inflammation in mesangial cells A ZFAS1 expression was significantly inhibited in the HG-induced cell, after silencing ZFAS1. Silencing ZFAS1 significantly inhibited the proliferation of the HG-induced cell compared to the negative control group ( B ). Oxidative stress induced after ZFAS1 knockdown in HG-treated HGMC. The level of ROS and MDA were significantly reduced in the HG-treated cell relative to the HG-si-NC group while the SOD activity significantly increased ( C ). The western blot analysis result showed that the protein expression level of fibronectin and collagen type IV was significantly inhibited in the HG si-ZFAS relative to the control HG si-NC group ( D ). The level of inflammatory cytokines IL-6, IL-1β, and TNF-α, produced by HG-induced cells after ZFAS1 knockdown. The inflammatory cytokines were significantly downregulated after ZFAS1 knockdown ( E ). All the experimental data are shown as mean ± SD of three independent experiments and the significance level was defined as p < 0.05 (** p < 0.01 and *** p < 0.001 vs. normal group; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. HG-si-NC group)

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Negative Control, Knockdown, Activity Assay, Western Blot, Control, Produced

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: ZFAS1 endogenously targets the miR-588 sequence. Bioinformatics prediction of target miRNA. Lncbase predicted that the ZFAS1 targets miR-588 seed region sequences. Dual-luciferase reporter assay experiment. MiR-588 mimics significantly reduced the luciferase activity of HGMC co-transfected with the ZFAS1 wild type (WT) reporter plasmid but had no inhibitory effect on that of ZFAS1 mutant type (Mut) plasmid ( A ). Biotinylated RNA pull-down assay. Biotin-coupled probes with wild type (WT) miR-588 sequence pull-down more ZFAS1 than those with the mutated miR-588 sequence (MUT) ( B ). RIP-qRT-PCR analysis. More ZFAS1 and miR-588 were significantly enriched in the immunoprecipitated Ago2-containing complexes compared to the IgG group ( C ). Experiments were carried out in triplicates and p < 0.05 was chosen as the significance level (*** p < 0.001 and ### p < 0.001)

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Sequencing, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Pull Down Assay, Quantitative RT-PCR, Immunoprecipitation

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: MiR-588 regulates HG-induced proliferation, oxidative stress, fibrosis, and inflammation in mesangial cells. The expression level of miR-588 in DN and HG-induced HGMC was significantly downregulated compared to the healthy or control group and was inversely related to ZFAS1 expression ( A – C ). CCK-assay and western blot analysis revealed the overexpression of miR-588 in HG-induced HGMC significantly reduced the cell viability, and the fibronectin and collagen type IV protein expression level, compared to the negative control group ( D and E ). overexpression of miR-588 markedly reversed the HG-induced upregulation of ROS and MDA, and downregulation of SOD, in the HG miR-NC treatment group ( F ). The overexpression of miR-588 significantly reduced the level of the HG-induced inflammatory cytokines secreted ( G ). Significance level was defined as p < 0.05 (** p < 0.01 and *** p < 0.001 vs. normal group; ## p < 0.01, and ### p < 0.001 vs. HG-si-NC group)

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Control, Western Blot, Over Expression, Negative Control

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: ROCK1 mRNA was a direct target of miR-588. Targetscan prediction of miR-588 target mRNA. Dual-luciferase reporter experiments. MiR-588 could significantly inhibit the luciferase activity of the HGMC co-transfected with the wild type (WT) ROCK reporter plasmid but had no inhibiting effect on that of the mutant type (MUT) ROCK group ( A ). QRT-PCR and Western blot analysis. Inhibiting miR-588 markedly upregulated the mRNA and protein expression level of ROCK in the HGMC compared to the negative control group while the overexpression of miR-588 reversed this ( B ). Inhibiting miR-588 restored si-ZFAS1-downregulated expression of ROCK1 mRNA and protein in the HGMC cell ( C ). (*** p < 0.001 ### p < 0.001)

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Quantitative RT-PCR, Western Blot, Expressing, Negative Control, Over Expression

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA ZFAS1 regulates the proliferation, oxidative stress, fibrosis, and inflammation of high glucose-induced human mesangial cells via the miR-588/ROCK1 axis

doi: 10.1186/s13098-022-00791-3

Figure Lengend Snippet: ZFAS1 inhibits HG-induced proliferation, oxidative stress, fibrosis, and inflammation in mesangial cells via the miR-588/ROCK1 axis. Transfecting pcDNA-ROCK1 overexpression plasmid into HGMC significantly upregulated ROCK1 expression ( A ). The overexpression of ROCK1 restored HG-induced cell viability that was reduced by the depleted ZFAS1 ( B ). MiR-588 inhibition or overexpression of ROCK1 partly restored the depleted level of ROS and MDA and repressed the SOD activity that was increased by ZFAS1 knockdown ( C ). MiR-588 inhibitor or overexpressed ROCK1 also recovered the downregulated fibronectin and collagen type IV protein expression that was induced by ZFAS1 knockdown in the HGMC ( D ). The depleted level of IL-6, IL-1β, and TNF-α after ZFAS1 knockdown in the HG-induced cell was also recovered by miR-588 inhibition or ROCK1 overexpression ( E ). Significance level was set at p < 0.05 (** p < 0.01, *** p < 0.001; ### p < 0.001)

Article Snippet: Human glomerular mesangial cells (HGMC) were bought from Sciencell Research Laboratories (Carlsbad, CA, USA).

Techniques: Over Expression, Plasmid Preparation, Expressing, Inhibition, Activity Assay, Knockdown

Journal: Bioscience Reports

Article Title: SNHG15 knockdown inhibits diabetic nephropathy progression in pediatric patients by regulating the miR-141/ICAM-1 axis in vitro

doi: 10.1042/BSR20204099

Figure Lengend Snippet: ( A ) The expression of miR-141 in HG-stimulated HGMCs was determined by quantitative reverse-transcription PCR (qRT-PCR); *** P <0.001 vs . the normal glucose (NG) group. ( B ) The expression of SNHG15 after transfection of miR-141 mimics/negative control (NC) or miR-141 inhibitor/inhibitor NC into HG-stimulated HGMCs was detected by qRT-PCR. ( C ) The viability of HG-stimulated HGMCs was measured by MTT assay. ( D ) The levels of IL-1β, IL-6 and TNF-α in HG-stimulated HGMCs were measured by ELISA. *** P <0.001 vs . the miR-NC group; ### P <0.001 vs . the inhibitor NC group.

Article Snippet: The proteins from HG-induced HGMCs were extracted using RIPA buffer containing protease inhibitor, followed by detecting the protein concentrations by a BCA Protein Assay Kit (Abcam).

Techniques: Expressing, Reverse Transcription, Quantitative RT-PCR, Transfection, Negative Control, MTT Assay, Enzyme-linked Immunosorbent Assay

Journal: Bioscience Reports

Article Title: SNHG15 knockdown inhibits diabetic nephropathy progression in pediatric patients by regulating the miR-141/ICAM-1 axis in vitro

doi: 10.1042/BSR20204099

Figure Lengend Snippet: ( A ) The predicted complementary binding site of ICAM-1 and miR-141. ( B ) The luciferase activity in human glomerular mesangial cells (HGMCs) co-transfected with pGL3-ICAM-1 WT/pGL3-ICAM-1 MUT and miR-141 mimics/negative control (NC) was determined by dual luciferase reporter (DLR) assay. *** P <0.001 vs. the miR-NC group. ( C ) The protein level of ICAM-1 was measured by Western blot assay. *** P <0.001 vs. the miR-NC group; ### P <0.001 vs. the inhibitor NC group.

Article Snippet: The proteins from HG-induced HGMCs were extracted using RIPA buffer containing protease inhibitor, followed by detecting the protein concentrations by a BCA Protein Assay Kit (Abcam).

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Negative Control, Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA KCNQ1OT1 affects cell proliferation, apoptosis and fibrosis through regulating miR-18b-5p/SORBS2 axis and NF-ĸB pathway in diabetic nephropathy

doi: 10.1186/s13098-020-00585-5

Figure Lengend Snippet: Knockdown of KCNQ1OT1 inhibited proliferation and fibrosis and promoted apoptosis in high glucose-induced HGMCs and HRGECs. High glucose-induced HGMCs and HRGECs were transfected with si-KCNQ1OT1. a The expression of KCNQ1OT1 was detected by qRT-PCR. b , c proliferation of HGMCs and HRGECs was determined by MTT assay. d apoptosis of HGMCs and HRGECs was determined by flow cytometry assay. Apoptotic rate: Annexin positive cells/all cells. e The expression of fibrosis-related proteins was detected by western blot assay in glucose-induced HGMCs and HRGECs. N = 3, * P < 0.05

Article Snippet: Human glomerular mesangial cells (HGMCs) and human renal glomerular endothelial cells (HRGECs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Knockdown, Transfection, Expressing, Quantitative RT-PCR, MTT Assay, Flow Cytometry, Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA KCNQ1OT1 affects cell proliferation, apoptosis and fibrosis through regulating miR-18b-5p/SORBS2 axis and NF-ĸB pathway in diabetic nephropathy

doi: 10.1186/s13098-020-00585-5

Figure Lengend Snippet: Silencing of SORBS2 repressed proliferation and facilitated apoptosis in high glucose-induced HGMCs and HRGECs. High glucose-induced HGMCs and HRGECs were transfected with si-SORBS2. a , b The mRNA and protein levels of SORBS2 were detected by qRT-PCR and western blot. c , d proliferation of HGMCs and HRGECs was determined by MTT assay. e apoptosis of HGMCs and HRGECs was determined by flow cytometry assay. f The expression of fibrosis-related proteins was measured by western blot assay. N = 3, * P < 0.05

Article Snippet: Human glomerular mesangial cells (HGMCs) and human renal glomerular endothelial cells (HRGECs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Transfection, Quantitative RT-PCR, Western Blot, MTT Assay, Flow Cytometry, Expressing

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA KCNQ1OT1 affects cell proliferation, apoptosis and fibrosis through regulating miR-18b-5p/SORBS2 axis and NF-ĸB pathway in diabetic nephropathy

doi: 10.1186/s13098-020-00585-5

Figure Lengend Snippet: KCNQ1OT1 affected proliferation and apoptosis by regulating SORBS2 expression in high glucose-induced HGMCs and HRGECs. a , b the mRNA level of SORBS2 was detected by qRT-PCR in HGMCs and HRGECs transfected with si-KCNQ1OT1 or pcDNA-KCNQ1OT1. c , d the protein level of SORBS2 was detected by western blot in HGMCs and HRGECs transfected with si-KCNQ1OT1 or pcDNA-KCNQ1OT1. e the protein level of SORBS2 was detected in HGMCs and HRGECs transfected with pcDNA-SORBS2. f , g , proliferation of HGMCs and HRGECs was determined after transfected with si-KCNQ1OT1 or si-KCNQ1OT1 + pcDNA-SORBS2. h and i apoptosis of HGMCs and HRGECs was determined after transfected with si-KCNQ1OT1 or si-KCNQ1OT1 + pcDNA-SORBS2. j western blot assay was utilized to detect the fibrosis-related proteins. N = 3, * P < 0.05

Article Snippet: Human glomerular mesangial cells (HGMCs) and human renal glomerular endothelial cells (HRGECs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA KCNQ1OT1 affects cell proliferation, apoptosis and fibrosis through regulating miR-18b-5p/SORBS2 axis and NF-ĸB pathway in diabetic nephropathy

doi: 10.1186/s13098-020-00585-5

Figure Lengend Snippet: KCNQ1OT1 targeted miR-18b-5p to regulate SORBS2 expression. a the putative binding sites between KCNQ1OT1 and miR-18b-5p were predicated by starBase 3.0. b The luciferase activities of WT-KCNQ1OT1 and MUT-KCNQ1OT1 were measured in 293 T cells transfected with miR-NC or miR-18b-5p. c The expression of miR-18b-5p was detected by qRT-PCR in HGMCs and HRGECs transfected with pcDNA-KCNQ1OT1. d The expression of miR-18b-5p was detected by qRT-PCR in HGMCs and HRGECs transfected with miR-18b-5p inhibitor. e , f proliferation of HGMCs and HRGECs was determined after transfected with si-KCNQ1OT1 or si-KCNQ1OT1 + miR-18b-5p inhibitor. g , h apoptosis of HGMCs and HRGECs was determined after transfected with si-KCNQ1OT1 or si-KCNQ1OT1 + miR-18b-5p inhibitor. i the fibrosis-related proteins were detected by western blot after transfection with si-KCNQ1OT1 or si-KCNQ1OT1 + miR-18b-5p inhibitor. j The putative binding sites between miR-18b-5p and SORBS2 were predicated by starBase 3.0. k The luciferase activities of WT-SORBS2 and MUT- SORBS2 were measured in 293 T cells transfected with miR-NC or miR-18b-5p. l , m , the mRNA and protein levels of SORBS2 were detected in HGMCs and HRGECs transfected with miR-18b-5p or miR-18b-5p + pcDNA-KCNQ1OT1. N = 3, * P < 0.05

Article Snippet: Human glomerular mesangial cells (HGMCs) and human renal glomerular endothelial cells (HRGECs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Expressing, Binding Assay, Luciferase, Transfection, Quantitative RT-PCR, Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: LncRNA KCNQ1OT1 affects cell proliferation, apoptosis and fibrosis through regulating miR-18b-5p/SORBS2 axis and NF-ĸB pathway in diabetic nephropathy

doi: 10.1186/s13098-020-00585-5

Figure Lengend Snippet: Knockdown of KCNQ1OT1 inhibited EMT and NF-ĸB pathway. a , b the mRNA and protein levels of Twist, NF-κB and STAT3 were detected in HGMCs transfected with si-KCNQ1OT1. c , d the mRNA and protein levels of Twist, NF-κB and STAT3 were detected in HRGECs transfected with si-KCNQ1OT1. N = 3, * P < 0.05

Article Snippet: Human glomerular mesangial cells (HGMCs) and human renal glomerular endothelial cells (HRGECs) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA).

Techniques: Knockdown, Transfection