Journal: International Journal of Molecular Sciences

Article Title: Resveratrol Pretreatment Ameliorates Concanavalin A-Induced Advanced Renal Glomerulosclerosis in Aged Mice through Upregulation of Sirtuin 1-Mediated Klotho Expression

doi: 10.3390/ijms21186766

Figure Lengend Snippet: Effects of resveratrol and klotho gene silencing on concanavalin A (Con A)-induced changes in the protein levels of klotho and SIRT-1, glomerulosclerosis-regulated factors, and reactive oxygen species (ROS) in mesangial cells. ( A ) The mRNA levels of transforming growth factor (TGF)-β, fibronectin, procollagen-III, and collagen I, and ( B ) the relative ROS levels were detected using flow cytometry after preincubation of mesangial cells with 25 μM resveratrol for 2 h, followed by cotreatment with 25 μΜ resveratrol plus 20 μg/mL Con A for 30 min. ( C ) mRNA levels of TGF-β, fibronectin, procollagen-III and collagen I, and ( D ) ROS relative levels were measured using flow cytometry after sequential treatments with klotho siRNA, 25 μΜ resveratrol for 2 h, 25 μΜ resveratrol plus 20 μg/ml Con A for 30 min. Different letters (a, b, and c) indicate significant differences between the groups ( p < 0.05).

Article Snippet: The mouse glomerular mesangial cell line was obtained from the Bioresource Collection and Research Center, Food Industry Research and Development Institute (cat. No. 60366; Hsinchu, Taiwan).

Techniques: Flow Cytometry

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