Acalypha indica root extract increases post-hypoxic rat hippocampal tissue culture cell viability via phospholipase A2 inhibition.

Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, and inhibition of its activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level was determined using ELISA. Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15, and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs 7.96 ng/mL, p < 0.05). Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL.

Acalypha indica Linn root extract improved hippocampal cell viability and increased brain-derived neurotrophic factor (BDNF) in hypoxic condition.

Background: This study was done to determine the effect of root extract of Acalypha indica Linn (akar kucing) in protecting neuron viability of the rat hippocampus on tissue culture in hypoxic condition. Methods: This is an experimental study of in vitro primary cell culture of hippocampus of Sprague Dowley adult rat. The cultures were group into control (C) and exposure to root extract of Acalypha indica Linn with dose of 10 mg/mL, 15 mg/mL, and 20 mg/mL for 72 hours. The cultures were then exposed to hypoxic gas (5% oxygen, 5% carbondioxide, nitrogen balance) for 24 hours. After that, relative cell viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), cell proliferation by 5-bromo2’-deoxy-uridine (BrdU), and Brain-derived Neurotrophic Factor (BDNF) levels by BDNF ELISA kit. Results: The result showed MTT viability (C: 99.7%, A indica L 10: 326.3%, A indica L 15: 411.7%, A indica L 20: 445.9%), BrdU absorbance (C: 0.07, A indica L 10: 0.10, A indica L 15: 0.12, A indica L 20: 0.13) of the exposured hippocampal cell were significantly higher than the control group (p < 0.01) accompany by increased level of BDNF (C: 11.3 pg/mL, A indica L 10: 12.5 pg/mL, A indica L 15: 23.1 pg/mL, A indica L 20: 18.1 pg/mL). Conclusion: The root extract of Acalypha indica Linn is able to improve rat hippocampal cell viability and endogenous BDNF levels in hypoxic condition.

Increased cell viability and proliferation in post-hypoxic hippocampal tissue culture treated with Acalypha indica root extract.

Background: This research was done to study the influence of Acalypha indica Linn root extract towards relative cell viability and proliferation as parameters of neurogenesis in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from adult Sprague Dawley rat exposed to hypoxia with 5% O2/5% CO2/N2 balance gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to 3 treatment groups. No treatment was given to the control group. Each group consists of 6 samples. After 90 hours of incubation, relative cell viability was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and cell proliferation was measured by using 5-bromo2’-deoxy-uridine (BrdU) for cell proliferation. Data was analyzed using one way ANOVA parametric tests, then further analyzed with post-hoc analysis. Results: The relative cell viability of rat hippocampal tissue culture treated with Acalypha indica Linn root extract with dose of 10, 15, and 20 mg/mL was significantly higher than control (176.95%, 220.62%, and 386.02% vs. 100%). Cell proliferation of rat hippocampal tissue culture treated with Acalypha indica Linn root extract with dose of 10, 15, and 20 mg/mL was significantly higher than control (0.132, 0.117, 0.114 vs 0.096). Conclusion: Acalypha indica Linn root extract with doses of 10, 15, and 20 mg/mL can increase relative cell viability and proliferation in post-hypoxic hippocampal tissue culture.

The nerve protection and in vivo therapeutic effect of Acalypha indica extract in frogs.

Aim To demonstrate nerve protection and/or treatment effect of Acalypha indica Linn. extract on nerve paralysis induced by subcutaneus injection of pancuronium bromide on frog’s back. Methods The study was performed on sixty frogs (Bufo melanostictus Schneider) that divided into two groups, i.e. the neuro-protection and neuro-therapy group. Each group was divided further into 6 sub-treatment groups: negative control group treated by water and positive control group treated by piracetam, treatment groups received the extracts 200, 300, 400, 500 mg/kgBW. Pancuronium bromide 0.2% (1 : 20 dilutions) were injected subcutaneously as muscle relaxant. The protective effect was studied by giving the extract orally, 1 hour prior to injection; while the therapeutic effect of the extract was studied by 10 minute treatment after injecting pancuronium bromide solution. The parameters measured were the onset and duration of paralysis (in minutes) and the recovery time (time needed to recover into normal condition). Results The study showed signifi cantly different protective effect of Acalypha indica Linn. root water extract at 400 and 500 mg/Kg.BW compared to negative control group and positive control group (piracetam (p<0.05); while the therapeutic effect was obvious at the dose 200-500 mg/Kg.BW compared to negative control group (p=0.000). There was no signifi cant difference compared to positive control group (piracetam), except at 300 mg/Kg.BW (p=0,012). Conclusion These results have proven that the water extract of Acalypha indica Linn. root has comparable protective and treatment effect on nerves system, as piracetam, but further studies should be performed to provide more evidences particularly pharmacokinetic and pharmacodynamic studies on two animal models that commonly used.