动物造模 z-bio 2022-09-26 652

　　Objective: To establish a method to evaluate the effect of paeoniflorin on the concentration of free Ca2+ in rat dorsal root ganglion neurons.

　　Methods: Rat dorsal root ganglia (DRG) were obtained by microdissection, digested with trypsin, sieved, and cultured and purified alternately with DF-12 and anti-mitotic medium to obtain primary rat DRG neurons. The purity of DRG neurons was determined by immunofluorescence; laser confocal microscopy was used to observe the change of intracellular Ca2+ fluorescence intensity, and the change rate of Ca2+ fluorescence intensity was analyzed to explore the effect of paeoniflorin on the concentration of free calcium ions in DRG cells and Effects of capsaicin receptors.

　　Results: The purity of DRG cells isolated by the above method can be as high as 95%. Capsaicin can inhibit the increase of intracellular Ca2+ by blocking the effect of capsaicin-activated transient receptor potential channel. Paeoniflorin exhibits similar performance to capsaicin. It can block the influx of extracellular Ca2+.

　　Conclusion: Paeoniflorin may inhibit the large increase of Ca2+ in DRG cells by acting on TRPV1 channel. This method can be used to evaluate the effect of drugs on Ca2+ concentration in rat DRG cells.