Withaferin A is a steroidal lactone that is isolated from the plant Withania somnifera. It has potent anti-inflammatory properties as it inhibits the activation of NF-κ B signaling pathway. The anti-tumor activity of Withaferin A is due to its ability to alter cytoskeletal architecture by binding annexin II and disrupting F actin cross-links. Withaferin A also inhibits angiogenesis by binding to vimentin and F-actin.
Withaferin A/醉茄素A 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Human retinal microvascular endothelial cells (HRMECs)
50 µM
15 minutes
To investigate the protective effect of Withaferin A on HRMECs under ischemia-reperfusion (I/R) injury. Results showed that Withaferin A reduced I/R-induced apoptosis of HRMECs by upregulating the expression of antioxidant molecules HO-1 and Prdx-1.
Cells. 2022 Oct 2;11(19):3113.
HaCaT cells
1 µM
15 minutes
To assess the effect of Withaferin-A on keratin filament dynamics. Results showed that Withaferin-A significantly reduced fluorescence recovery, indicating slowed turnover of keratin filaments.
Int J Mol Sci. 2020 Jun 23;21(12):4450.
Primary peritoneal macrophage
0.2 and 0.5 µM
2 hours
To test the direct anti-inflammatory effect of WA on LPS-induced inflammatory response, results showed WA at 0.2 and 0.5 μM dose-dependently attenuated LPS-induced upregulation of Tnfa, Il6, Il1b mRNAs.
Cell Death Dis. 2021 Feb 11;12(2):174.
BV2/NF-κB-Luc cells
2.5 µM
2 hours
To assess the effect of WFA on NF-κB activity in BV2/NF-κB-Luc cells. Results showed that WFA significantly reduced LPS-induced NF-κB activity.
Neurotherapeutics. 2021 Jan;18(1):286-296.
MCF7 ARE-luciferase reporter cells
1 µM
20 hours
To evaluate the effect of WA on ARE-luciferase reporter activity. WA significantly increased luciferase activity, indicating activation of ARE via the Nrf2 pathway.
Free Radic Biol Med. 2016 Dec;101:116-128.
Wild-type mouse embryonic fibroblasts (WT MEF)
0.1, 0.3, 0.7, 1 µM
20 hours
To evaluate the induction of Nqo1 and Ho-1 transcripts by WA. WA dose-dependently induced Nqo1 and Ho-1 transcripts in WT MEF but not in Nrf2-disrupted MEF.
Free Radic Biol Med. 2016 Dec;101:116-128.
NSC-34/NF-κB-Luc cells
1 µM
20 minutes
To assess the effect of WFA on NF-κB activity in NSC-34/NF-κB-Luc cells. Results showed that WFA significantly reduced TNF-α-induced NF-κB activity.
Neurotherapeutics. 2021 Jan;18(1):286-296.
SH-SY5Y neuroblast cells
1, 10, 100 nM
24 hours
To evaluate the protective effect of WFA against hemin-induced cell injury, results showed that WFA significantly increased cell viability and reduced the level of oxidative stress marker MDA, while increasing the activities of anti-oxidative stress markers GSH-Px and SOD.
Neural Regen Res. 2023 Jun;18(6):1308-1315.
SH-SY5Y cells
2 µM
24 hours
To evaluate the protective effect of WA against MPP+-induced neurotoxicity, results showed WA improved cell viability.
Cell Death Differ. 2021 Aug;28(8):2517-2535.
Bladder cancer J82 cells
0 to 10 µM
24 hours
To evaluate the antiproliferative effect of WFA on bladder cancer J82 cells. Results showed that WFA reduced cell viability in a dose-dependent manner with an IC50 value of 3.1 μM.
Antioxidants (Basel). 2021 Jun 30;10(7):1063.
Human Epidermal Keratinocytes (HEKs)
0.5-3 µM
3 hours
To evaluate the effect of Withaferin-A on keratin filament assembly. Results showed that Withaferin-A disrupts keratin filaments in a dose-dependent manner, starting at 0.5 µM with a 5-fold reduction at 3 µM.
Int J Mol Sci. 2020 Jun 23;21(12):4450.
Primary mouse hepatocytes
0.01–0.5 µM
30 minutes
To test the direct anti-apoptotic effect of WA on ACTD/TNF-α and GalN/TNF-α-induced apoptosis models, results showed WA at 0.01–0.5 μM had no significant effect in reducing cell death.
Cell Death Dis. 2021 Feb 11;12(2):174.
THP-1 cells
20 µM
4 hours
To investigate the regulatory effect of WFA on inflammatory cytokines/chemokines. Results showed that WFA treatment significantly downregulated 38 cytokines/chemokines (e.g., IL-1β, GM-CSF, CCL2/MCP-1) and upregulated 3 (RLN2, TNFRSF8/CD30, MPO).
Front Immunol. 2018 Feb 9;9:195.
HA1800 cells
1 or 3 µM
48 hours
To evaluate the effects of WA on the viability of GBM cells and normal astrocytes. WA inhibited the growth of U87 and U251 cells more than normal astrocyte cells HA1800 at the same concentration.
Cell Prolif. 2020 Jan;53(1):e12706.
U251 cells
1 or 3 µM
48 hours
To evaluate the effects of WA on the viability of GBM cells and normal astrocytes. WA inhibited the growth of U87 and U251 cells more than normal astrocyte cells HA1800 at the same concentration.
Cell Prolif. 2020 Jan;53(1):e12706.
U87 cells
1 or 3 µM
48 hours
To evaluate the effects of WA on the viability of GBM cells and normal astrocytes. WA inhibited the growth of U87 and U251 cells more than normal astrocyte cells HA1800 at the same concentration.
Cell Prolif. 2020 Jan;53(1):e12706.
Human cardiomyocyte cell line AC16
0.5 µM
48 hours
To investigate the protective effect of Withaferin A on cardiomyocytes under acute hypoxia, results showed that WA pretreatment significantly reduced apoptosis and necrosis of cardiomyocytes.
Cells. 2022 Dec 25;12(1):85.
AML12 hepatocytes
0.01–1.0 µM
6 hours
To evaluate the protective effect of Withaferin A against H2O2-induced oxidative stress and necrosis. Results showed that Withaferin A significantly enhanced cell survival, reduced LDH release, and decreased oxidative stress.
Biochem Pharmacol. 2015 Sep 1;97(1):122-32.
SH-SY5Y cells
2 μM
24 hours
To evaluate the protective effect of WA against MPP+-induced neurotoxicity, results showed that WA significantly increased cell viability.
Cell Death Differ. 2021 Aug;28(8):2517-2535
Withaferin A/醉茄素A 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6J mice
ICH model
Intracerebroventricular injection
0.1, 1, 5 μg/kg
Once daily for 7 days
To evaluate the neuroprotective effect of WFA after ICH, results showed that WFA significantly reduced brain tissue injury and iron deposition, and improved neurological function in a dose-dependent manner.
Neural Regen Res. 2023 Jun;18(6):1308-1315.
Mice
Diet-induced obese (DIO) mice
Intraperitoneal injection
1.25-2 mg/kg
Once daily for 21 days
Evaluate the weight-reducing effect of Withaferin A as a leptin sensitizer, results showed DIO mice had significant body weight reduction (22.8%), decreased food intake (62%), reduced fat mass (35%), and complete resolution of hepatic steatosis
Nat Med. 2016 Sep;22(9):1023-32.
C57Bl/6J mice
Retinal ischemia-reperfusion injury model
Intraperitoneal injection
10 nM/g
Single dose 8 hours before surgery
To investigate the protective effect of Withaferin A on retinal ischemia-reperfusion injury in vivo. Results showed that Withaferin A activated the Akt signaling pathway, upregulated the expression of HO-1 and Prdx-1, and reduced retinal I/R injury and apoptosis of HRMECs.
Cells. 2022 Oct 2;11(19):3113.
Sprague-Dawley (SD) rats
Simulated extreme hypoxic environment at 7620 m
Intraperitoneal injection
2 mg/kg
Once daily for 7 days
To investigate the protective effect of Withaferin A on SD rats in an extreme hypoxic environment, results showed that WA pretreatment significantly improved the 24-hour survival rate, reduced myocardial damage, and maintained cardiac function.
Cells. 2022 Dec 25;12(1):85.
Mice
Liver tumor model
Intraperitoneal injection
2.5 mg/kg
2 weeks
WFA monotreatment prolonged median survival by 44.5 days, and the combination with α-PD-1 and SB225002 significantly improved survival rates
Gut. 2023 Sep;72(9):1774-1782
Mice
MPTP-induced Parkinson's disease model
Intraperitoneal injection
20 μg/kg
Once daily for 7 days
To evaluate the protective effect of WA on dopaminergic neuron loss and motor deficits, results showed WA alleviated neuron loss and motor deficits.
Cell Death Differ. 2021 Aug;28(8):2517-2535.
Mice
PTEN conditional knockout mouse model
Oral
3 mg/kg and 5 mg/kg
Daily administration for 45 weeks
To evaluate the chemopreventive effect of WA on prostate cancer in PTEN-deficient mice. WA significantly inhibited primary tumor growth, completely suppressed metastatic lesions, and downregulated pAKT expression and EMT markers.
Neoplasia. 2017 Jun;19(6):451-459
C57BL/6 mice
Angiotensin II-induced cardiac cachexia model
Intraperitoneal injection
4 mg/kg
Once every three days for three weeks
WFA treatment significantly improved Ang II-induced left ventricular dysfunction and cardiac hypertrophy, reduced cardiac fibrosis and inflammatory responses
Cells. 2024 May 3;13(9):783
C57BL/6J mice
Angiotensin II-induced muscle cachexia model
Intraperitoneal injection
4 mg/kg
Once every three days for three weeks
Withaferin A attenuates muscle cachexia induced by Angiotensin II through regulating pathways activated by Angiotensin II, restoring muscle mass and strength.
Cells. 2025 Feb 8;14(4):244
C57BL/6NTac male mice
Acetaminophen (APAP)-induced liver injury model
Intraperitoneal injection
40 mg/kg
Single dose, euthanized at 4 h and 16 h after APAP injection
To evaluate the protective effect of Withaferin A against APAP-induced liver injury. Results showed that Withaferin A significantly reduced serum ALT levels, hepatocyte necrosis, and intrahepatic hemorrhage, decreased GSH depletion, JNK activation, mitochondrial Bax translocation, and nitrotyrosine generation, and upregulated the expression of Nrf2, Gclc, and Nqo1, while downregulating Il-6 and Il-1β.
Biochem Pharmacol. 2015 Sep 1;97(1):122-32.
Nude mice
U87 xenograft model
Tail vein injection
5 mg/kg
Once daily for 27 days
To evaluate the inhibitory effect of WA on the growth of U87 xenografts. The tumor volume and weight in the WA-treated group were significantly smaller than those in the control group.
Cell Prolif. 2020 Jan;53(1):e12706.
Mice
TDP-43G348C transgenic mouse model
Intraperitoneal injection
5 mg/kg
Once every 2 days for 8 weeks
To evaluate the effects of WFA on cognitive function and TDP-43 pathology in TDP-43G348C transgenic mice. Results showed that WFA treatment improved cognitive function, reduced TDP-43 cytoplasmic aggregation and neuroinflammation, and increased levels of the autophagic marker LC3BII.
Neurotherapeutics. 2021 Jan;18(1):286-296.
C57BL/6J mice
Wild-type, Nrf2-disrupted, Nrf2flox/flox, and Nrf2flox/flox::AlbCre mice
Oral gavage
7 mg/kg
Single dose, euthanized 20 hours later
To evaluate the protective effect of WA against APAP-induced hepatotoxicity and its Nrf2-dependence. WA significantly protected wild-type mice from APAP hepatotoxicity but not Nrf2-disrupted mice.
Free Radic Biol Med. 2016 Dec;101:116-128.
C57BL/6 mice
MPTP-induced Parkinson's disease mouse model
Intraperitoneal injection
20 μg/kg
Once daily for 7 days
To evaluate the protective effect of WA against MPTP-induced Parkinson's disease symptoms, results showed that WA significantly alleviated the loss of dopaminergic neurons and motor deficits.
搜索
