Journal: bioRxiv

Article Title: Remdesivir inhibits renal fibrosis in obstructed kidneys

doi: 10.1101/2020.04.01.019943

Figure Lengend Snippet: GS-441524, the intermediate metabolite of remdesivir, inhibited renal fibrosis in vitro . (A) Following TGF-β stimulation, renal fibroblasts NRK-49F cells were treated with various concentration (0, 0.1, 1, 10, 100 μM) of GS-441524 for 24 hours. CCK8 assay was performed to determine the viability of NRK-49F cells. (B and C) The expression of fibronectin (FN), phosphorylated Smad3 (pSmad3), and alpha-smooth muscle actin (α-SMA) of NRK-49F cells were analyzed by Western blotting and quantified; (D) Upon TGF-β stimulation, human renal epithelial HK2 cells were treated with various concentration (0, 0.1, 1, 10, 100 μM) of GS-441524 for 48 hours. CCK8 assay was performed to determine the viability of HK2 cells. (E and F) The expression of FN, pSmad3, and α-SMA of HK2 cells were analyzed by Western blotting and quantified. Data represent mean ± SD. * P < 0.05 versus Vehicle-DMSO; ** P < 0.01 versus Vehicle-DMSO; # P < 0.05 versus TGF-β-DMSO; ## P < 0.01 versus TGF-β-DMSO. One representative result of at least three independent experiments is shown.

Article Snippet: Remdesivir (Product name GS-5734; Cat. No. CSN19703) was purchased from CSNpharm (Chicago, Illinois, USA) and dissolved in DMSO as a 50mg/ml stock, which was further diluted into normal saline by sonication as a working solution.

Techniques: In Vitro, Concentration Assay, CCK-8 Assay, Expressing, Western Blot

Journal: bioRxiv

Article Title: Remdesivir inhibits renal fibrosis in obstructed kidneys

doi: 10.1101/2020.04.01.019943

Figure Lengend Snippet: Remdesivir inhibited renal fibrosis in vitro . (A) Following TGF-β stimulation, renal fibroblasts NRK-49F cells were treated with various concentration (0, 0.01, 0.1, 1, 10 μM) of remdesivir for 24 hours. CCK8 assay was performed to determine the viability of NRK-49F cells. (B and C) The expression of FN, pSmad3, and α-SMA of NRK-49F cells were analyzed by Western blotting and quantified; (D) Upon TGF-β stimulation, human renal epithelial HK2 cells were treated with various concentration (0, 0.01, 0.1, 1, 10 μM) of remdesivir for 48 hours. CCK8 assay was performed to determine the viability of HK2 cells. (E and F) The expression of FN, pSmad3, and α-SMA of HK2 cells were analyzed by Western blotting and quantified. Data represent mean ± SD. * P < 0.05 versus Vehicle-DMSO; ** P < 0.01 versus Vehicle-DMSO; # P < 0.05 versus TGF-β-DMSO; ## P < 0.01 versus TGF-β-DMSO. One representative result of at least three independent experiments is shown.

Article Snippet: Remdesivir (Product name GS-5734; Cat. No. CSN19703) was purchased from CSNpharm (Chicago, Illinois, USA) and dissolved in DMSO as a 50mg/ml stock, which was further diluted into normal saline by sonication as a working solution.

Techniques: In Vitro, Concentration Assay, CCK-8 Assay, Expressing, Western Blot

Journal: bioRxiv

Article Title: Remdesivir inhibits renal fibrosis in obstructed kidneys

doi: 10.1101/2020.04.01.019943

Figure Lengend Snippet: Intraperitoneal (i.p.) administration of remdesivir inhibited fibrosis in UUO mice. After sham or UUO operation, wide type c57 mice were treated with 10 mg/kg/d remdesivir by i.p. injection for 10 days. Serum and kidney tissue were collected 1 hour after remdesivir injection at day 10. (A) Renal fibrosis was assessed by Masson’s trichrome staining. (B) The expression of FN, collagen I (Col-1), pSmad3, and α-SMA by Western blotting. One representative of at least three independent experiments is shown. (C) Liver function (ALT and AST) and renal function (Scr and BUN) were assessed. (D) The concentrations of nucleoside metabolite (GS-441524) and alanine metabolite (Ala-Met), two remdesivir metabolites, in serum and kidneys were determined by LC-MS/MS. Data represent mean ± SD. ND represents not determined. * P < 0.05 versus Sham-vehicle; ** P < 0.01 versus Sham-vehicle; # P < 0.05 versus UUO-vehicle; ## P < 0.01 versus UUO-vehicle.

Article Snippet: Remdesivir (Product name GS-5734; Cat. No. CSN19703) was purchased from CSNpharm (Chicago, Illinois, USA) and dissolved in DMSO as a 50mg/ml stock, which was further diluted into normal saline by sonication as a working solution.

Techniques: Injection, Staining, Expressing, Western Blot, Liquid Chromatography with Mass Spectroscopy

Journal: bioRxiv

Article Title: Remdesivir inhibits renal fibrosis in obstructed kidneys

doi: 10.1101/2020.04.01.019943

Figure Lengend Snippet: Intrarenal administration of remdesivir inhibited fibrosis in UUO mice. 50μL of vehicle or remdesivir (1 mg/mL) was injected intrarenally to the left kidney, which was subjected to the UUO operation thereafter. Serum and renal tissues were collected at 1 hour or day7 after UUO operation. (A) Renal fibrosis was assessed by Masson’s trichrome staining. (B) The expression of FN, collagen I (Col-1), pSmad3, and α-SMA by Western blotting. One representative of at least three independent experiments is shown. (C) Liver function (ALT and AST) and renal function (Scr and BUN) were assessed. (D) The concentrations of nucleoside metabolite (GS-441524) and alanine metabolite (Ala-Met), two remdesivir metabolites, in serum and kidney were determined by LC-MS/MS. Data represent mean ± SD. ND represents not determined. # P < 0.05 versus UUO-vehicle; ## P < 0.01 versus UUO-vehicle.

Article Snippet: Remdesivir (Product name GS-5734; Cat. No. CSN19703) was purchased from CSNpharm (Chicago, Illinois, USA) and dissolved in DMSO as a 50mg/ml stock, which was further diluted into normal saline by sonication as a working solution.

Techniques: Injection, Staining, Expressing, Western Blot, Liquid Chromatography with Mass Spectroscopy