Anti-metastatic potential of Punarnavine, an alkaloid from Boerhaavia diffusa Linn.

The alkaloid Punarnavine isolated from the plant Boerhaavia diffusa Linn. was studied for its anti-metastatic activity using B16F-10 melanoma cells in C57BL/6 mice. Administration of Punarnavine (40mg/kg body weight) prophylactically (95.25%), simultaneously (93.9%) and 10 days after tumor inoculation (80.1%) could inhibit the metastatic colony formation of melanoma in lungs. Survival rate of the metastatic tumor-bearing animals were increased significantly by the administration of Punarnavine in all the modalities compared to the metastasis bearing untreated control. These results correlated with the biochemical parameters such as lung collagen hydroxyl proline, uronic acid, hexosamine, serum sialic acid, serum gamma-glutamyltranspeptidase and serum vascular endothelial growth factor (VEGF) levels and histopathological studies. Punarnavine administration could suppress or down regulate the expression of MMP-2, MMP-9 (MMP--matrix metalloproteinase), ERK-1, ERK-2 (ERK--extracellular-signal-regulated kinase) and VEGF in the lung tissue of metastasis-induced animals. Punarnavine could inhibit MMP-2 and MMP-9 protein expression in gelatin zymographic analysis of B16F-10 cells. These results indicate Punarnavine could inhibit the metastatic progression of B16F-10 melanoma cells in mice.

Boerhaavia diffusa stimulates cell-mediated immune response by upregulating IL-2 and downregulating the pro-inflammatory cytokines and GM-CSF in B16F-10 metastatic melanoma bearing mice.

Effect of Boerhaavia diffusa extract on the Cell Mediated Immune (CMI) response in metastatic condition was studied using C57BL/6 mice model. Administration of Boerhaavia diffusa enhanced Natural Killer (NK) cell activity, antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent complement mediated cytotoxicity (ACC) and the activity was observed in treated group much earlier compared to the metastatic tumor bearing control. Production of the cytokine IL-2 was significantly enhanced by the administration of Boerhaavia diffusa compared to the untreated metastatic tumor bearing control. Levels of GM-CSF and pro-inflammatory cytokines such as IL-1beta, IL-6 and TNF-a were significantly lowered by Boerhaavia diffusa administration compared to metastatic control. The gene expression level of IL-2, IL-1beta, IL-6, TNF-alpha and GM-CSF in B16F-10 cells also correlate the above result. These results indicate Boerhaavia diffusa could enhance the immune response against metastatic progression of B16F-10 melanoma cells in mice.

Ursolic acid induces apoptosis by activating p53 and caspase-3 gene expressions and suppressing NF-kappaB mediated activation of bcl-2 in B16F-10 melanoma cells.

The objective of this study was to assess the effect of ursolic acid, a triterpene on inducing apoptosis in B16F-10 melanoma cells. Treatment of B16F-10 cells with nontoxic concentration of ursolic acid showed the presence of apoptotic bodies and induced DNA fragmentation in a dose depended manner. The apoptotic genes p53 and caspase-3 were found to be upregulated while the anti-apoptotic gene bcl-2 was down regulated in ursolic acid treated cells. The transcription factors NF-kappaBp65, NF-kappaBp50, NF-kappaBc-Rel, c-FOS, ATF-2 and CREB-1 were found to be inhibited significantly (p<0.001) in ursolic acid treated cells compared to untreated control. The pro-inflammatory cytokine production and gene expression of TNF-alpha, IL-1beta, IL-6 and GM-CSF were down regulated in ursolic acid treated cells compared to nontreated B16F-10 metastatic melanoma cells. All these results demonstrate that ursolic acid induce apoptosis via inhibition of NF-kappaB induced bcl-2 mediated anti-apoptotic pathway and subsequent activation of p53 mediated and TNF-alpha induced caspase-3 mediated pro-apoptotic pathways.

Studies on the protective effects of Boerhaavia diffusa L. against gamma radiation induced damage in mice.

The radioprotective effect of the hydro-alcoholic extract of Boerhaavia diffusa was studied using the in vivo mice model. The sublethally irradiated mice (600 rads, single dose) were treated intraperitoneally with 20 mg/kg of the extract. The animals were sacrificed at different time periods after the whole-body radiation. The most affected tissues--bone marrow and intestine--were considerably protected by the intraperitoneal administration of B. diffusa as estimated by bone marrow cellularity, maturing monocytes, and intestinal glutathione. Total white blood cell count was lowered drastically after radiation exposure (ninth day, 1500+/-500 cells/ mm(3)). When the animals were exposed to radiation and treated with B. diffusa, the total white blood cell count was lowered only to 4000+/-400 cells/mm(3) on the third day, and it reached an almost normal level (6250+/-470 cells/mm(3)) by the ninth day. The elevated level of serum and liver alkaline phosphatase after radiation exposure was reduced in the B. diffusa-treated group. The serum and liver glutamate pyruvate transferase, which were elevated after radiation exposure, were also reduced by treatment with B. diffusa compared to the control. The lipid peroxidation level also increased in the irradiated animals both in the liver and serum, but in B. diffusa-treated animals, there was a significant reduction in lipid peroxidation levels. The agarose gel electrophoresis of DNA isolated from bone marrow of mice exposed to gamma radiation showed heavy damage that was reduced by treatment with B. diffusa. These results are indicative of the radioprotective effect of the whole-plant extract of B. diffusa.

Effect of Punarnavine, an alkaloid from Boerhaavia diffusa, on cell-mediated immune responses and TIMP-1 in B16F-10 metastatic melanoma-bearing mice.

Effect of Punarnavine on the cell-mediated immune (CMI) response in metastatic condition was studied using C57BL/6 mice model. Administration of Punarnavine enhanced Natural Killer (NK) cell activity, antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent complement mediated cytotoxicity (ACC) and the activity was observed in treated group much earlier compared to the metastatic tumor-bearing control. Production of cytokines such as IL-2 and IFN-gamma were significantly enhanced by the administration of Punarnavine compared to the untreated metastatic tumor-bearing control. Peaks of pro-inflammatory cytokines such as IL-1beta, IL-6 and TNF-alpha were significantly lowered by Punarnavine administration compared to metastatic control. The level and expression of TIMP-1 was also enhanced by the administration of Punarnavine compared to metastatic tumor bearing control. These results indicate Punarnavine could enhance the immune response against metastatic progression of B16F-10 melanoma cells in mice.