| Object i veTo observe the influence of serum containing Asarum on the voltage gated sodium channel in isolated SD rat's medulla DRG neurons.Methods1. Make the blood serum containing Asarum.First prepare certain Asarum root powder of density give SD rat to irritated stomach, twice a day,for eight days.The dosage of Asarum were according to The Pharmacopeia of the People's republic of China(1st part, public in 2005).2. The influence of serum containing Asarum on the INa in isolated SD rat's medulla DRG neurons.Single rat medulla DRG neurons were acutely isolated by enzymatic digestion and mechanical dispersion from 1-3-day-old SD rat. Briefly, 400-500 mm thick brain slices were cut from medulla .Then the medulla DRG region was removed from the slice and incubated for 60min at 32°C in artificial cerebrospinal solution(ACSF), bubbled with 9 5%O2+5%CO2; and successively transferred into D-hanks containing trypsin at 32°C for 15 min. The brain fragments collected from several brain slices were treated by gentle pipetting using a fine glass tube.Whole-cell patch clamp recordings were made with EPC-10 double amplifier after the neurons were transferred to 35 mm culture dish filled with 1. 5-2ml ACSF with pipettes resistance of 1-5MΩ. Cells were held at a holding potential (HP) of -80mV, a series of 100 ms depolarizing steps to -80mV- +30mV increment at 1OmV each step and then stored in computer using pulse+pulsefit (version8.74) software.For drug application, a series of ten micro-tubes (200 μM) were glued together side by side. Solutions were fed from independent reservoirs by gravity. The microtubes were shifted horizontally with a micro-manipulator for aligning the flow of solution from the tubes relative to the cells.ResultsThe normal serum can't increased the peak of INa in isolated SD rat's medulla DRG neurons. The peak of INa change from 1.121±... |
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