ABSTRACT
Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into various cell types such as cartilage, bone, and fat cells. Recent studies have shown that induction of MSCs in vitro by growth factors including epidermal growth factor (EGF) and fibroblast growth factor (FGF2) causes them to differentiate into neural like cells. These cultures also express ChAT, a cholinergic marker; and TH, a dopaminergic marker for neural cells. To establish a protocol with maximum differentiation potential, we examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP-9, retinoic acid, and heparin. Adipose-derived MSCs were extracted and expanded in vitro for 3 passages after reaching >80% confluency, for a total duration of 9 days. Cells were then characterized by flow cytometry for CD markers as CD44 positive and CD45 negative. MSCs were then treated with neural induction media and were characterized by morphological changes and Q-PCR. Differentiated MSCs expressed markers for immature and mature neurons; ß Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons. Improved protocols for MSCs induction will facilitate and ensure the reproducibility and standard production of MSCs for therapeutic applications in neurodegenerative diseases.
Subject(s)
Adipose Tissue/cytology , Cholinergic Neurons/physiology , Dopaminergic Neurons/physiology , Mesenchymal Stem Cells/physiology , Neural Stem Cells/physiology , Neurogenesis , Adult , Cell Lineage , Cell Separation , Cells, Cultured , Cholinergic Neurons/drug effects , Cholinergic Neurons/metabolism , Culture Media/metabolism , Dopaminergic Neurons/drug effects , Dopaminergic Neurons/metabolism , Female , Gene Expression Profiling , Gene Expression Regulation, Developmental , Humans , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Microtubule-Associated Proteins/metabolism , Neural Stem Cells/drug effects , Neural Stem Cells/metabolism , Neurogenesis/drug effects , Phenotype , Tubulin/metabolismABSTRACT
Achillea millefolium Linn. (Asteraceae) is used in folk medicine for the treatment of overactive cardiovascular and respiratory ailments. This study describes its hypotensive, cardio-depressant, vasodilatory and bronchodilatory activities. The crude extract of Achillea millefolium (Am.Cr) caused a dose-dependent (1-100 mg/kg) fall in arterial blood pressure of rats under anaesthesia. In spontaneously beating guinea-pig atrial tissues, Am.Cr exhibited negative inotropic and chronotropic effects. In isolated rabbit aortic rings, Am.Cr at 0.3-10 mg/mL relaxed phenylephrine (PE, 1 µm) and high K(+) (80 mm)-induced contractions, as well as suppressed the PE (1 µm) control peaks obtained in Ca(++) -free medium, like that caused by verapamil. The vasodilator effect of Am.Cr was partially blocked by N(ω) -nitro-l-arginine methyl ester in endothelium intact preparations. In guinea-pig tracheal strips, Am.Cr inhibited carbachol (CCh, 1 µm) and K(+) -induced contractions. These results indicate that Achillea millefolium exhibits hypotensive, cardiovascular inhibitory and bronchodilatory effects, thus explaining its medicinal use in hyperactive cardiovascular and airway disorders, such as hypertension and asthma.
Subject(s)
Achillea/chemistry , Blood Pressure/drug effects , Bronchodilator Agents/pharmacology , Cardiovascular System/drug effects , Plant Extracts/pharmacology , Animals , Female , Guinea Pigs , In Vitro Techniques , Male , Rabbits , Rats , Rats, Sprague-DawleyABSTRACT
Taxus wallichiana Zucc. (Himalayan Yew) is often used in northern areas of Pakistan for the treatment of pyrexia, acute pains and epilepsy. We have investigated certain pharmacological activities of the methanol leaf extract against convulsion, nociception and pyrexia induced in rodents. The aim was to justify and explore its folk uses in these pathological conditions, on scientific basis. The studies were carried out using acetic acid-induced nociception and pentylenetetrazole-induced convulsions in mice, while formalin test and yeast-induced pyrexia in rats. Significant analgesic (67.77 and 74.29%) effect was found in acetic acid-induced model at doses of 100 and 200mg/kg, i.p. respectively. Crude extract exhibited significant (P<0.05) inhibition of the formalin noxious stimulation on both early and late phases of pain by the extracts (100 and 200mg/kg doses). In case of yeast-induced pyrexia model, 200mg/kg dose showed very significant (P<0.01) inhibition while 50 and 100mg/kg dose caused a significant (P<0.05) inhibition. Plant extract has controlled the pentylenetetrazole-induced convulsions in mice. 100 and 200mg/kg i.p doses of the extract significantly (P<0.05) inhibited the mioclonus and clonus while inhibition of tonus and hind limb tonic extension (HLTE) was highly significant (P<0.01). The anticonvulsant activity of this plant has been reported for the first time throughout the whole genus. The observed pharmacological activities provide the scientific basis for the folkloric use of the plant in treating epilepsy, pyrexia and acute pain.
