Construction and preliminary functional study of heterologous expression system of TRPV1 channel / 中国药理学通报

Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.

TRPV1 channel-mediated thermogenesis is a common mode for the Chinese pungent-hot or pungent-warm herbs to demonstrate their natures / 药学学报

To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.