The Effect Of Mahuang Lianqiao Chixiaodou Decoction Drug Serum On The Electrophysiology Of The Rat Isolated DRG Neurons In The Rat Model

ObjectivePreliminary experimental study found Mahuang Lianqiao Chixiaodou Decoction(MLC)can improve allergic inflammation.Mahuang Lianqiao Chixiaodou Decoction(MLC)in treatment of cholestasis itching will be further studied.The clinical treatment of cholestasis itching curative effect is distinct,but the related mechanism of treatment is not clear.The TRPV1 channel expressed in the dorsal root nerve is taken as the breakthrough point of research,and its mechanism of action is explored to lay the foundation for further study.MethodsIn this study,we used the surgical ligation of rat bile duct to make the rat model of cholestasis.Behavioral observation was conducted after the rat model.The drug serum was prepared by using high performance liquid chromatography-mass spectrometry(HPLC-MS)method.The experiment was divided into a blank serum group,5% drug serum group,20% of the serum group.By the acute separation of the rat dorsal root neurons(DRG)cells,it can be go to patch clamp experiments.The TRPV1 channel was activated by capsaicin,and the effect of drug serum on the channel current was observed.Results(1)Behavioral measurement:the scratch behavior of rats with different degrees began to appear in the week after the mold-making,and there was a significant difference between the scratch behavior of rats and the behavior of blank rats through behavioral test(P<0.05).(2)HPLC-MS test:there is a small amount of forsythia in the drug containing serum.(3)Acute isolation and culture of DRG neurons in rats:it was complete with smooth edges and good wall.(4)Patch clamp detection:capsaicin can activate the TRPV1 channel current of the DRG cells,when the serum containing the drug alone acted on the cells,it was found that the containing serum can not cause any current changes;Capsaicin and drug containing serum were used to superposition the cells,and it was found that there was a significant inhibitory effect on the TRPV1 channel activated by capsaicin;and the blank serum had no significant effect on the channel current(P>0.05).The 5% concentration serum was significantly different from the blank serum(P<0.05);the inhibitory effect of 20% concentration was stronger than the 5% concentration(P<0.05).Conclusions(1)By behavioral testing:the replication of the rat model of cholestasis was successful.(2)Quantitative analysis of the composition of forsythia in MLC containing serum by liquid mass,and the results showed that the preparation of MLC containing drug serum was successful;(3)Acute isolated rat DRG neurons and culture: through microscopic observation,it was demonstrated the success of cell culture;(4)The patch clamp experiment:Mahuang Lianqiao Chixiaodou Decoction(MLC)medicated serum by acting on TRPV1 channels can inhibit channel current,and reduce the internal flow of calcium ions,which may inhibit the release of the itch causing substances to achieve the purpose of the treatment of cholestasis itching.TRPV1 channels may be one of MLC in treatment of cholestasis itch signaling pathways.