Journal: iScience

Article Title: MircroRNA-92b as a negative regulator of the TGF-β signaling by targeting the type I receptor

doi: 10.1016/j.isci.2023.108131

Figure Lengend Snippet:

Article Snippet: Human renal glomerular endothelial cells (HRGECs) (ScienCell, Carlsbad, CA, USA; Cat. #4000) were cultured in Endothelial Cell Medium (ScienCell, Cat.#1001) and human mesangial mells (HMCs) (ScienCell, Cat. #4200) were cultured in Mesangial Cell Medium (ScienCell, Cat. #4201).

Techniques: Recombinant, Control, Plasmid Preparation, Software, Modification, Staining, Extraction, Bicinchoninic Acid Protein Assay, Western Blot, cDNA Synthesis, SYBR Green Assay, Immunohistochemistry, Isolation

Journal: Molecules

Article Title: Synthetic Epoxyeicosatrienoic Acid Mimics Protect Mesangial Cells from Sorafenib-Induced Cell Death

doi: 10.3390/molecules30071445

Figure Lengend Snippet: Cell viability of human renal mesangial cells measured by a WST assay ( N = 4). E 10µM : median increase in cell viability in percentage points upon treatment with EET analog (10 µM) relative to sorafenib (10 µM) only. E 1µM : EET-analog (1 µM). Missing entries denote insignificant ( p > 0.05, Student’s t ) increase in cell viability. HsEH : IC 50 for human soluble epoxide hydrolase (HsEH).

Article Snippet: Human renal mesangial cells (4200, ScienCell, Carlsbad, CA, USA) were cultured at 37 °C (in 5% CO2) in RPMI 1640 medium (GibcoTM, LS11875093) containing 10% FBS, 100 U/mL of penicillin, and 0.1 mg/mL of streptomycin.

Techniques: WST Assay

Journal: Molecules

Article Title: Synthetic Epoxyeicosatrienoic Acid Mimics Protect Mesangial Cells from Sorafenib-Induced Cell Death

doi: 10.3390/molecules30071445

Figure Lengend Snippet: WST-8 assay of human renal mesangial cells treated with sorafenib and 8,9-EET mimics, ( a ) 7b , ( b ) 8b , ( c ) 10b , ( d ) 8d , at indicated concentrations after 48 h incubation, as percentages relative to control. Bars represent mean ± SEM ( n = 4).

Article Snippet: Human renal mesangial cells (4200, ScienCell, Carlsbad, CA, USA) were cultured at 37 °C (in 5% CO2) in RPMI 1640 medium (GibcoTM, LS11875093) containing 10% FBS, 100 U/mL of penicillin, and 0.1 mg/mL of streptomycin.

Techniques: Incubation, Control

Journal: Viruses

Article Title: Expression Profile of Human Renal Mesangial Cells Is Altered by Infection with Pathogenic Puumala Orthohantavirus

doi: 10.3390/v14040823

Figure Lengend Snippet: Receptor expression in primary human mesangial cells. Surface expression of hantaviral receptor proteins integrin α v β 3 and CD55 was analyzed in two donors by flow cytometry. CHO cells served as negative control and Vero E6 cells as positive control for integrin α v β 3 expression. Upper panels: isotype controls, lower panels: receptor staining.

Article Snippet: Primary human renal mesangial cells (HRMC, donor #5948 and #20018) were obtained from Sciencell and maintained in mesangial cell medium (Sciencell, Carlsbad, CA, USA).

Techniques: Expressing, Flow Cytometry, Negative Control, Positive Control, Staining

Journal: Viruses

Article Title: Expression Profile of Human Renal Mesangial Cells Is Altered by Infection with Pathogenic Puumala Orthohantavirus

doi: 10.3390/v14040823

Figure Lengend Snippet: Analysis of functional consequences in PUUV-infected mesangial cells. Viability ( A ), migration capacity ( B ) and adhesion ( C ) of PUUV-infected HRMCs (4 dpi) were analyzed. Infection was quantified by staining of N protein. Results of uninfected cells were set to 100%. Three independent experiments were performed. The mean ± SD is shown. ns: not significant.

Article Snippet: Primary human renal mesangial cells (HRMC, donor #5948 and #20018) were obtained from Sciencell and maintained in mesangial cell medium (Sciencell, Carlsbad, CA, USA).

Techniques: Functional Assay, Infection, Migration, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: The high expression of circ-ACTR2 was detected in DN samples and HG-treated HRMCs. A , B The qRT-PCR was performed to detect the circ-ACTR2 level in normal and DN samples ( A ) or control and HG-treated HRMCs ( B ). C The determination of circ-ACTR2 and linear ACTR2 was conducted by qRT-PCR after treatment of RNase R in total RNA. D The circ-ACTR2, U6 and GAPDH levels were assayed by qRT-PCR in cytoplasm and nucleus. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HG-induced cell damages in HRMCs were partly reversed by the silence of circ-ACTR2. A The expression of circ-ACTR2 was quantified using qRT-PCR in four groups of control, HG, HG + si-NC, HG + si-ACTR2 in HRMCs. B Cell viability detection was performed by CCK-8 assay. C Inflammatory cytokines IL-6 and TNF-α were determined by ELISA. D Cell proliferation analysis was performed by EdU assay. E The protein examination of collagen I and collagen IV was conducted by western blot. F , G SOD activity ( F ) and MDA level ( G ) were respectively measured using the corresponding kits. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 exhibited the sponge function of miR-205-5p. A Starbase v2.0 predicted the binding sites between circ-ACTR2 and miR-205-5p. B The miR-205-5p level was examined by qRT-PCR after transfection of miR-NC or miR-205-5p. C , D Dual-luciferase reporter assay ( C ) and RIP assay ( D ) were used to confirm whether circ-ACTR2 combined with miR-205-5p. E The quantification of miR-205-5p was carried out using qRT-PCR in normal and DN tissues. F The linear analysis between circ-ACTR2 and miR-205-5p was performed using the Pearson’s correlation coefficient. G The effect of HG on the miR-205-5p expression was analyzed via qRT-PCR. H The level of circ-ACTR2 was detected by qRT-PCR in control, HG, HG + pcD5-ciR or HG + circ-ACTR2 group. I The regulation of circ-ACTR2 inhibition or overexpression on the miR-205-5p expression was performed by qRT-PCR in HG-treated HRMCs. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Binding Assay, Quantitative RT-PCR, Transfection, Luciferase, Reporter Assay, Expressing, Control, Inhibition, Over Expression

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2/miR-205-5p axis affected the regulation of HG in HRMCs. A The transfection efficiency of anti-miR-205-5p was assessed by qRT-PCR. B The qRT-PCR was applied for the expression detection of miR-205-5p after transfection of si-NC, si-ACTR2, si-ACTR2 + anti-miR-NC or si-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. C Cell viability was examined via CCK-8 assay. D The concentrations of IL-6 and TNF-α were tested via ELSIA. E Cell proliferation was assessed via EdU assay. F , G The levels of collagen I and collagen IV were assayed via western blot. H , I Oxidative stress was evaluated by SOD activity ( H ) and MDA level ( I ) using the kits. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HMGA2 downregulation protected against the HG-induced cell dysfunction in HRMCs. A The protein expression of HMGA2 in control, HG, HG + si-NC or HG + si-HMGA2 group was evaluated by western blot. B CCK-8 was performed to analyze cell viability. C ELISA was performed to measure the concentrations of IL-6 and TNF-α. D EdU assay was conducted to determine cell proliferation. E , F Western blot was conducted to detect the protein levels of collagen I and collagen IV. G , H SOD activity ( G ) and MDA level ( H ) by the detection kits were exploited to examine the oxidative stress. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Control, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 regulated the HMGA2 level by acting as a sponge of miR-205-5p. A , B The mRNA ( A ) and protein ( B ) levels of HMGA2 were respectively detected by qRT-PCR and western blot after transfection of si-NC, si-circ-ACTR2, si-circ-ACTR2 + anti-miR-NC or si-circ-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Quantitative RT-PCR, Western Blot, Transfection

Journal: International Journal of Molecular Sciences

Article Title: The Ethyl Acetate Extract of Phyllanthus emblica L. Alleviates Diabetic Nephropathy in a Murine Model of Diabetes

doi: 10.3390/ijms25126686

Figure Lengend Snippet: The targeted gene effects of ethyl acetate extract of P. emblica L. (EPE) in human renal mesangial cell lysates by Western blot analyses with a primary antibody against TGF-β1 and Collagen IV. ( A ) Representative blots in human renal mesangial cells; ( B , C ) Quantification of the expression levels of TGF-β1 and Collagen IV. All values are means ± S.E. +++ p < 0.001 compared with the control group; * p < 0.05, *** p < 0.001 compared with the HG group. The β-actin was used as the internal standard in each sample. HG: high glucose. EPE: ethyl acetate extract of P. emblica L.

Article Snippet: Human renal mesangial cell (HRMC) (no. 4200; ScienCell Research Laboratories (Carlsbad, CA, USA) lysates were treated with 1 μg/mL, 5 μg/mL, or 10 μg/mL EPE for 24 h, and then, the cells were collected to examine the expression levels of TGF-β1 and collagen IV.

Techniques: Western Blot, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: The Ethyl Acetate Extract of Phyllanthus emblica L. Alleviates Diabetic Nephropathy in a Murine Model of Diabetes

doi: 10.3390/ijms25126686

Figure Lengend Snippet: Representative photographs of the kidney; ethyl acetate extract of P. emblica L. (EPE) in db/db mice on ( A ) glomerular basement membrane in cortex and ( B ) mesangial matrix expansion in outer medulla by periodic acid Schiff (PAS) staining (magnification: 20 × 10); ( C ) expression levels of nephrin in the kidneys by immunohistochemical (IHC) staining; ( D ) Nephrin-positive area (%) score by IHC stains compared with the db/db group. EPE: EPE1, EPE2, and EPE3 (100, 200, and 400 mg/kg body). ### p < 0.001 compared with the db/m group; *** p < 0.001 compared with the db/db + vehicle (distilled water) (db/db) group.

Article Snippet: Human renal mesangial cell (HRMC) (no. 4200; ScienCell Research Laboratories (Carlsbad, CA, USA) lysates were treated with 1 μg/mL, 5 μg/mL, or 10 μg/mL EPE for 24 h, and then, the cells were collected to examine the expression levels of TGF-β1 and collagen IV.

Techniques: Membrane, Staining, Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: The Ethyl Acetate Extract of Phyllanthus emblica L. Alleviates Diabetic Nephropathy in a Murine Model of Diabetes

doi: 10.3390/ijms25126686

Figure Lengend Snippet: Effects of seven fractions of EPE (EA) on expression levels of collagen IV, TGF-β1, VEGF, and KIM in human renal mesangial (HRM) cells by Western blotting analyses. Human renal mesangial (HRM) cells were treated with seven fractions and equal amounts of lysates were resolved by SDS-PAGE and blotted for collagen IV, TGF-β1, VEGF, and KIM. ( A – E ) Effects of 7 fractions of ethyl acetate extract of P. emblica L. (EtOAc soluble fractions) (EA-1, EA-2, EA-3, EA-A, EA-5, EA-6, and EA-7) on expression levels of Collagen IV, TGF-β1, KIM, and VEGF in HRM cells by Western blotting analysis. Human renal mesangial (HRM) cells were treated with 7 fractions as described in the experimental procedures and equal amounts of lysates were resolved by SDS-PAGE and blotted for collagen IV, TGF-β1, KIM, and VEGF. ( A ) Representative blots for 7 fractions in HRM cells; ( B – E ) Quantification of the expression levels to β-actin including ( B ) collagen IV, ( C ) TGF-β1, ( D ) KIM, and ( E ) VEGF. All values are means ± S.E. ### p < 0.001 compared with the control group; * p < 0.05, *** p < 0.001 compared with the glucose group. ( F , G ) High-performance liquid chromatography analysis of ( F ) 2500 ppm ethyl acetate of P. emblica L. (EPE), ( G ) 10.3 mg/5 mL EA-4 of ethyl acetate of P. emblica L.

Article Snippet: Human renal mesangial cell (HRMC) (no. 4200; ScienCell Research Laboratories (Carlsbad, CA, USA) lysates were treated with 1 μg/mL, 5 μg/mL, or 10 μg/mL EPE for 24 h, and then, the cells were collected to examine the expression levels of TGF-β1 and collagen IV.

Techniques: Expressing, Western Blot, SDS Page, Control, High Performance Liquid Chromatography