Oroxylum indicum, of the Bignoniaceae family, has various ethnomedical uses such as an astringent, anti-inflammatory, anti-bronchitis, anti-helminthic and anti-microbial, including anticancer properties. The druggability of OI stem bark extract was determined by its molecular docking interactions with PARP and Caspase-3, two proteins involved in cell survival and death. Note that 50 µg/mL of Oroxylum indicum extract (OIE) showed a significant (p < 0.05%) toxicity to HSC-3 cells. MTT aided cell viability and proliferation assay demonstrated that 50 µg/mL of OIE displayed significant (p < 0.5%) reduction in cell number at 4 h of incubation time. Cell elongation and spindle formation was noticed when HSC-3 cells were treated with 50 µg/mL of OIE. OIE initiated DNA breakage and apoptosis in HSC-3 cells, as evident from DNA ladder assay and calcein/EB staining. Apoptosis potential of OIE is confirmed by flow cytometer and triple-staining (live cell/apoptosis/necrosis) assay. Caspase-3/7 fluorescence quenching (LANCE) assay demonstrated that 50 µg/mL of OIE significantly enhanced the RFU of caspases-3/7, indicating that the apoptosis potential of OIE is probably through the activation of caspases. Immuno-cytochemistry of HSC-3 cells treated with 50 µg/mL of OIE showed a significant reduction in mitochondrial bodies as well as a reduction in RFU in 60 min of incubation time. Immunoblotting studies clearly showed that treatment of HSC-3 cells with OI extract caused caspase-3 activation and PARP deactivation, resulting in apoptotic cell death. Overall, our data indicate that OIE is an effective apoptotic agent for human squamous carcinoma cells and it could be a future cancer chemotherapeutic target.
Objectives: To evaluate the radio-mitigating property of Asparagus racemosus root ethanolic extract and Isoprinosine against ionizing radiationinduced harmful effects in Swiss albino mice. Methods: The experimental animals were exposed to sublethal dose (6 Gy) of whole-body electron beam radiation and administered orally with Asparagus racemosus root ethanolic extract (ARE-200mg/kg bd. wt) and Isoprinosine (IPR-400 mg/ kg bd. wt) for 15 consecutive days. After the experimental period, the animals were sacrificed. Blood and liver tissue were collected. Blood sample was used for assessment of hematological parameters, serum was used for biochemical analysis (lipid peroxidation, total antioxidant capacity) and the level of pro-inflammatory cytokines was determined in the liver homogenate. Results: The radiation control group showed a statistically significant reduction in the hematological parameters. IPR post-treatment group showed a substantial increase in the percentage of lymphocytes and monocytes. ARE and IPR post-treatment caused a statistically significant improvement in platelet count. Irradiation of mice resulted in a significant elevation in lipid peroxidation. Post-treatment of mice with ARE and IPR caused significant depletion in lipid peroxidation. The serum total antioxidant capacity (TAC) in the irradiated group decreased significantly. Administration of ARE to the animals after irradiation caused a significant elevation in TAC level. IPR post-treatment did not show any substantial change in the reduced level of TAC. The cytokines level in the liver homogenate of irradiated mice showed that Interleukin-2 (IL-2), Tumor Necrosis Factor alpha (TNF-α) and Interferon gamma (IFN-γ) significantly increased after radiation exposure. Administration of ARE and IPR after irradiation significantly reduced the levels of IL-2 and TNF-α in the posttreatment groups. An elevated level of IFN-γ was reduced considerably in the IPR post-treatment animals. Conclusion: Asparagus racemosus root ethanolic extract and Isoprinosine exhibit radio-mitigating effects against electron beam radiation-induced detrimental effects. This occurs through several mechanisms, such as free radical scavenging, anti-inflammation, facilitation of repair activity, regeneration of hematopoietic cells and affecting to molecular levels.
Objective: We aimed to evaluate anticlastogenic and radioprotective potential of Asparagus racemosus root extract (ARE) and Isoprinosine (IPR) against electron beam radiation (EBR) induced clastogenicity and toxicity in Swiss albino mice. Methodology: In the pre-radiation study, the experimental animals were orally administered ARE -200mg and IPR -400mg/ kg b.wt once daily for 15 consecutive days. The animals exposed to sublethal dose (6Gy) of whole body EBR. Chromosomal aberration analysis and micronucleus assay were carried out in the bone marrow cells of the experimental animals. The various types of aberrations were scored and the micronuclei in Polychromatic Erythrocytes (PCE) and Nomochromatic Erythrocytes (NCE) were recorded. Assessment of Granulocyte Macrophage Colony Stimulating Factor (GM-CSF), was performed using mouse GM-CSF Picokine ELISA kit. Non-specific Alpha -esterase activity was determined by simultaneous azo dye coupling method. Dose Reduction Factor (DRF) was calculated to determine the protective role of ARE and IPR against EBR. Result: Treatment of mice with ARE-200 mg/kg b.wt and IPR-400mg/kg b.wt decreased the percentage of the total aberration compared to the irradiated group; significantly reduced (P<0.05) the frequency of Mn PCE and Mn NCE when compared with irradiation alone groups. Irradiation reduced the level of GM-CSF in the splenocytes which was enhanced by the pre-treatment with ARE and IPR. There was a significant increase in the number of alpha-esterase positive cells in the pre-treatment group compared to radiation control. Increase in survival percentage was observed in the pre-treated mice when compared to radiation alone group. The DRF value of 1.11 and 1.04 was observed respectively. Conclusion:The present study suggests that the antioxidant potential of ARE and IPR could be of extreme significance in offering radioprotection and may be useful in combating various free-radical and reactive oxygen speciesmediated human pathological conditions.
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