human renal mesangial cell line hrmc  (BioVector NTCC)

Journal: Journal of Healthcare Engineering

Article Title: lncRNA MSC-AS1 Aggravates Diabetic Nephropathy by Regulating the miR-325/CCNG1 Axis

doi: 10.1155/2022/2279072

Figure Lengend Snippet: LncRNA MSC-AS1 knockdown alleviates HG-induced proliferation, fibrosis, and inflammation of HRMC cells. (a) The expression of MSC-AS1 in serum samples from DN patients and normal subjects ( n = 30). (b) The expression of MSC-AS1 in HG- and NG-treated HRMC cells. (c) The expression of MSC-AS1 in HG-treated HRMC cells with its siRNA. (d, e) Cell proliferation and apoptosis were detected in the NG, HG, and HG + si-MSC-AS1 groups. (f) Cell fibrosis was assessed by the levels of FN, Col IV, and Col I in the NG, HG, and HG + si-MSC-AS1 groups. (g) The release of TNF- α , IL-6, and IL-1 β was examined in the NG, HG, and HG + si-MSC-AS1 groups. ∗ P < 0.05 ∗∗ P < 0.01.

Article Snippet: Human renal mesangial cell line HRMC (Procell, Wuhan, China) was cultured in a matched dedicated complete medium (CM-H067; Procell) at 37°C with 5% CO 2 .

Techniques: Knockdown, Expressing

Journal: Journal of Healthcare Engineering

Article Title: lncRNA MSC-AS1 Aggravates Diabetic Nephropathy by Regulating the miR-325/CCNG1 Axis

doi: 10.1155/2022/2279072

Figure Lengend Snippet: lncRNA MSC-AS1 functions as a miR-325 sponge. (a) The binding sites between MSC-AS1 and miR-325. (b) The relationship between MSC-AS1 and miR-325 was verified by pull-down assay. (c) MiR-325 expression was detected in HG-treated HRMC cells with MSC-AS1 siRNA and vector. (d) MSC-AS1 expression was measured in HG-treated HRMC cells containing miR-325 mimics or inhibitors. (e) MiR-325 expression was detected in DN patients and normal subjects ( n = 30). (f) A negative correlation between MSC-AS1 and miR-325 expression was found in DN patients ( n = 30). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Human renal mesangial cell line HRMC (Procell, Wuhan, China) was cultured in a matched dedicated complete medium (CM-H067; Procell) at 37°C with 5% CO 2 .

Techniques: Binding Assay, Pull Down Assay, Expressing, Plasmid Preparation

Journal: Journal of Healthcare Engineering

Article Title: lncRNA MSC-AS1 Aggravates Diabetic Nephropathy by Regulating the miR-325/CCNG1 Axis

doi: 10.1155/2022/2279072

Figure Lengend Snippet: CCNG1 is a direct target of miR-325. (a) The binding sites between miR-325 and CCNG1. (b) The relationship between miR-325 and CCNG1 was confirmed by pull-down assay. (c) CCNG1 expression was measured in HG-treated HRMC cells containing miR-325 mimics or inhibitors. (d) CCNG1 expression was detected in DN patients and normal subjects. (e) A negative correlation between CCNG1 and miR-325 expression was identified in DN patients ( n = 30). (f) A positive correlation between MSC-AS1 and CCNG1 expression was detected in DN patients ( n = 30). (g) CCNG1 expression was assessed in HG-treated HRMC cells with MSC-AS1 siRNA and vector. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Human renal mesangial cell line HRMC (Procell, Wuhan, China) was cultured in a matched dedicated complete medium (CM-H067; Procell) at 37°C with 5% CO 2 .

Techniques: Binding Assay, Pull Down Assay, Expressing, Plasmid Preparation

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