体外研究 (In Vitro) | Amantadine sulfate (0-500 μM, 26 h) inhibits SARS-CoV-2 replication, with IC50concentrations between 83 and 119 μM[4]. Amantadine sulfate (0-100 μg/mL, 24-72 h) markedly inhibits the proliferation of HepG2 and SMMC-7721 cells[6]. Amantadine sulfate (0-75 μg/mL, 48 h) arrests the cell cycle at the G0/G1 phase and induces apoptosis[6]. Amantadine sulfate (0-75 μg/mL, 48 h) reduces the levels of the cell cycle-related genes and proteins (cyclin D1, cyclin E and CDK2), reduces Bcl-2 and increases the Bax protein and mRNA levels[6].
Cell Viability Assay[4] Cell Line: | Vero E6 cells | Concentration: | 500 μM, 100 μM, 20 μM, 4 μM, and 8 nM | Incubation Time: | 26 h | Result: | Caused a concentration-dependent reduction (IC50=83 μM) of viral nucleic acids in the supernatant 26 h after infection at 10-500 μM. Caused a concentration-dependent reduction (IC50=119 μM) of viral nucleic acids in the cytosol 26 h after infection. |
Cell Proliferation Assay[6] Cell Line: | Human HCC cell lines (HepG2 and SMMC-7721) and normal hepatocellular cells (L02 cells) | Concentration: | 0, 1, 2, 5, 10, 25, 50 and 100 μg/mL | Incubation Time: | 24, 48 and 72 h | Result: | Inhibited cellular proliferation in a time- and dose-dependent manner in HepG2 and SMMC-7721 cells. |
Cell Cycle Analysis[6] Cell Line: | HepG2 and SMMC-7721 cells | Concentration: | 0, 10, 25, 50 and 75 μg/mL | Incubation Time: | 48 h | Result: | Significantly increased the population of HepG2 and SMMC-7721 cells in the G0/G1 phase in a dose-dependent manner, and significantly decreased the number of HepG2 cells in the S phase. |
Apoptosis Analysis[6] Cell Line: | HepG2 and SMMC-7721 cells | Concentration: | 0, 10, 25, 50 and 75 μg/mL | Incubation Time: | 48 h | Result: | Markedly increased the percentage of apoptotic HepG2 and SMMC-7721 cells (early- and late-stage apoptosis) in a dose-dependent manner. |
Western Blot Analysis[6] Cell Line: | HepG2 and SMMC-7721 cells | Concentration: | 0, 10, 25, 50 and 75 μg/mL | Incubation Time: | 48 h | Result: | Showed downregulation of cyclin D1, cyclin E and CDK2, and showed a decrease in Bcl-2 levels and an increase of Bax levels in HepG2 and SMMC-7721 cells. |
RT-PCR[6] Cell Line: | HepG2 and SMMC-7721 cells | Concentration: | 0, 10, 25, 50 and 75 μg/mL | Incubation Time: | 48 h | Result: | Revealed an increase in Bax and decrease in Bcl-2 genes. |
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体内研究 (In Vivo) | Amantadine sulfate (25 mg/kg, IP, once daily for 3 days) inhibits surgery induced neuroinflammation and learning and memory impairment[5].
Animal Model: | Fischer 344 rats (Four-month old, male, 290-330 g, 15 rats each group)[5] | Dosage: | 25 mg/kg | Administration: | IP, once daily for 3 days (the first dose at 15 min before surgery) | Result: | Inhibited surgery induced neuroinflammation and learning and memory impairment, increased GDNF (glial cell line-derived neurotrophic factor) that was co-localized with glial fibrillary acidic protein (an astrocytic marker) in the hippocampus. |
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