Piperine attenuates the cancerous activity response in Neuro-2a cell line

Interactions of cancer cells with their microenvironment play a significant role in defining the severity of the disease. In search of novel compounds with anti-inflammatory and anticancerous capabilities, the effects of purified compound piperine were investigated in Neuro-2a cell line. The neuronal lineage of Neuro-2a cell line was confirmed by using antibody against Beta-III tubulin protein. The cells were treated with different concentrations of piperine [MuM: 10, 50 and 100] for 48 hrs at 37 degree C. A dose of 100 MuM was selected that induces a 50% inhibition in the cell growth calculated by MTT and morphometery assays. The result shows that in the presence of piperine neurite outgrowth was decreased in a dose dependent manner. The gene expression of TN-C, TNfnD and TnfnC were significantly reduced whereas the expression intensities of TnfnA1, TnfnA2, CSPGs and Laminin were significantly elevated when compared to their respective untreated controls. Similarly proinflammatory marker COX-2 expression was significantly inhibited in the presence of piperine when compared to untreated controls. This is the first time we have illustrated that irrespective of increased expressions of CSPGs, a significant reduction in Tenascin-C and its TNfnD and TNfnC domains are necessary to inhibit the tumor progression. Taken together, the capabilities of piperine to induce an apoptosis by decreasing the neurite outgrowth, proliferation rate and expression of TN-C and COX-2 in Neuro-2a cell line confirmed for its anticancerous and anti-inflammatory potential

Neurite outgrowth properties of calotropis procera: In search for a neuroprotectant

In view of the well-documented medicinal properties of Calotropis procera [CP], the present study was designed to evaluate the neuroprotective effect of the extract. We have prepared a methanolic extract of Calotropis procera and screen varying concentration of CP [20, 30, 40, 50 and 70microg/ml] for the stimulatory potency on neurite outgrowth. The stimulatory effect of CP on neurite outgrowth was assessed in primary hippocampal neurons. Neurite lengths were measured using optika provison analysis software. Neuritogenesis was further analyzed by immunostaining by using specific neuronal marker beta III-tubulin. The data show that neurite outgrowth from hippocampal neurons were significantly enhanced in the presence of CP [40microg/ml]. The most stimulatory neurite outgrowth effects were appeared after 48hrs incubation of neurons with CP [40microg/ml]. These data confirm that CP extract could promote invitro hippocampal neurite outgrowth in a dose-dependent manner. Our results indicate that CP can be used as a healthy dietary supplement for the cognitive functions of the brain

Vitex negundo induces an anticonvulsant effect by inhibiting voltage gated sodium channels in murine Neuro 2A cell line

Vitex negundo [Vn] extract is famous for the treatment of neurological diseases such as migraine and epilepsy. These neurological diseases have been associated with abnormally increased influx of sodium ions into the neurons. Drugs that inhibit voltage gated sodium channels can be used as potent anti-epileptics. Till now, the effects of Vn on sodium channels have not been investigated. Therefore, we have investigated the effects of methalonic fraction of Vn extract in Murine Neuro 2A cell line. Cells were cultured in a defined medium with or without the Vn extract [100 microg/ml]. Sodium currents were recorded using whole-cell patch clamp method. The data show that methanolic extract of Vn inhibited sodium currents in a dose dependent manner [IC50 =161microg/ml]. Vn [100 microg/ml] shifted the steady-state inactivation curve to the left or towards the hyper polarization state. However, Vn did not show any effects on outward rectifying potassium currents. Moreover, Vn [100 microg/ml] significantly reduced the sustained repetitive [48 +/- 4.8%, P<0.01] firing from neonatal hippocampal neurons at 12 DIV. Hence, our data suggested that inhibition of sodium channels by Vn may exert pharmacological effects in reducing pain and convulsions

Neuroprotective capabilities of Vitex negundo in primary hippocampal neurons

Vitex negundu [Vn] is a well-known aromatic shrub commonly used as a traditional folk medicine famous for its potential pharmacological and biological activities. Several chemical compounds are extracted and identified from the different parts of the Vn such as leaves, root, seeds and flowers. Number of researches reported the herb as antimicrobial, anti-androgenic, anti-osteoporotic, and anti-tumour, anti-cancer, anti-inflammatory, anti-oxidant, anti-hyperglycemic and hepatoprotective. The effects of Vn on neurite outgrowth have not been identified till now. Therefore present study was designed to investigate the neurite outgrowth effects of Vn extract in hippocampal neurons. Neurons from P0 mice were isolated and cultured in defined medium containing the different concentrations of Vn [20, 30, 40, 50, 100, 150 and 200 microg/ml] for 48 hrs. The presence of the neurites was confirmed by using betaIII-tubulin antibody which specifically labels only the neurites. Morphometric analysis was done by using Optika Pro-Vision software. The data show that Vn at 30 and 40 microg/ml significantly increased the mean average length of the longest neurite whereas at 150 and 200 microg/ml it significantly decreased the mean average length of the 10 longest neurite in hippocampal neurons. Nevertheless Vn did not show any significant effects on the sum of all the neurite lengths at any concentrations tested. Taken together the result shows that methanolic extract of Vn has potential to produce long neurites at 30 and 40 microg/ml and therefore can be act as a neuroprotective agent in the future drug development

Cardamom extract induces cell proliferation by increasing potassium currents in NIH3T3 cell line

Amommum subulatum [Roxb.] or Cardamom extract is known to have anti-inflammatory and neuroprotective effects towards many gastrointestinal related problems. However, uptill now different fractions of cardamom extract on fibroblasts with respect to potassium channel activity have not been investigated. Therefore, present study investigated the effects of different tractions of cardamom extract on potassium channels in non-tumor NIH3T3 cell line. Phytochemical analysis of hydroalcoholic, n-hexane, butane and ethyl acetate fractions of cardamom extracts were purified and isolated by thin layer chromatography [TLC]. 3T3 cells were cultured and incubated with hydroalcohol [1-2 Hg/ml], n-hexane [1 microg/ml], butane [2 microg/ml] and ethyl acetate [1-2 microg/ml] for 5 hrs at 37°C. Modulation in potassium currents were recorded by whole-cell patch clamp method. The data showed two constituents Cineol [CioHigO] and Terpinyl acetate [CioHi[7]OOCCH[3]] by TLC method. The present study shows that the constituents in n-hexane, hydro alcohol [1 [microg/ml] and ethyl acetate [2 microg/ml] significantly increased [p<0.01] the potassium outward rectifying currents from NIH3T3 cells when compared to untreated controls cells. Where as, butanol fraction [2 microg/ml] significantly decreased [p<0.01] the inward rectifying currents when compared to controls. Moreover hydroalcoholic and n-hexane fractions have increased the proliferation in 3T3 cell line. On the other hand butanol and ethyl acetate did not induce proliferation in 3T3 cells. Taken together, our data suggested that cardamom extract contains constituents that increased K[+] currents, cell migration and proliferation and are involved in wound healing