| Objective: Previous studies have been found that Mahuang Lianqiao Chixiaodou Decoction is used to treat various pruritic diseases of the skin,and it has a good curative effect.The mechanism of Mahuang Lianqiao Chixiaodou Decoction in the treatment of itching caused by cholestasis and the neurophysiological mechanism of the itching in cholestasis will be further observed.So the itching of cholestasis is taken as the breakthrough point,by the model rats which were made by the method of bile duct ligation,and the Mahuang Lianqiao Chixiaodou Decoction was administered,which to lay the foundation for further study the antipruritic mechanism of Mahuang Lianqiao Chixiaodou Decoction.Methods:1.Theoretical researchTo review the relevant information and comb through the understanding of pruritus in traditional Chinese medicine,and to analyze the mechanism of the antipruritic effect of Mahuang Lianqiao Chixiaodou Decoction.2.Experimental research72 SPF wistar rats were divided into six groups,and each group with 12 examples: the control group,conventional feeding.(2)the sham group,the rats in the sham group were only conducted in the abdominal incision suture,no bile duct ligation,and after the suture will awaken its own,after a day of fasting except water,namely began regular feed;(3)the model group,(4)lower dose treatment group,(5)middle dose treatment group,(6)high dose treatment group,the rats of the above four groups were conducted by bile duct ligation,after the surgery will awaken its own,after a day of fasting except water,start regular feeding,three treatment groups in three weeks after the operation began filling and taking different doses(low dose groups: 0.255 g/ml,dose group of 0.51 g/ml,high dose group of 1.02 g/ml)of Mahuang Lianqiao Chixiaodou Decoction,treatment time is two weeks,every record in the process of Mahuang Lianqiao Chixiaodou Decoction of general situation and the behavior of rats.By immunofluorescence double labeling labeling PAR2 antigen and antigen of TRPV1,detection of DRG neurons PAR2 and TRPV1 antigen expression,using real-time fluorescent quantitative PCR method to detect DRG neurons PAR2 mRNA and TRPV1 mRNA expression.Results:(1)The theoretical research indicates that itch is mainly related to wind,humidity,heat,insect,poison and other factors,among which,it is dominated by the wind,and can be combined with the evil,forming rheumatism,wind and heat,rheumatic fever and etc.(2)the behavior change the scratching behavior of the rats in model group compare with control group were significantly increasing,after the treatment,the scratching behavior of the rats in low dose group and middle dose group decreased moderately,in high dose group decreased more significantly,suggest the treatment of MLCD to pruritus in cholestasis have certain effect.(3)Immunofluorescence double testing results rats in the model group PAR2 antigen expression compared with the control group and sham group had significant difference(P<0.05),the treatment groups compared with the model group,low dose group and middle dose group had no significant difference(P>0.05),high dose group had significant difference(P<0.05).Rats in model group TRPV1 antigen expression compared with the control group and sham group had significant difference(P<0.05),the treatment groups compared with the model group,low dose group had no significant difference(P>0.05),middle dose group and high dose group had significant difference(P<0.05).(4)RT-PCR test results rats in the model group PAR2 mRNA expression of relative quantity compared with the control group and sham group had significant difference(P<0.05),the treatment groups compared with the model group,low dose group,middle dose group and high dose group had no significant difference(P>0.05).Rats in the model group TRPV1 mRNA expression of relative quantity compared with the control group and the sham group had significant difference(P<0.05),the treatment groups compared with the model group,low dose group and middle dose group had no significant difference(P>0.05),high dose group had significant difference(P<0.05),suggest that after conducting bile duct ligation,relative expression of TRPV1 mRNA in rats increased,and Mahuang Lianqiao Chixiaodou Decoction can effectively reduce the TRPV1 mRNA expression,the curative effect in high dose group of is the most significant in this experiment.Conclusions:1.The appearance of pruritus is closely related to wind,humidity,heat and other evil spirits,and the Mahuang Lianqiao Chixiaodou Decoction has a hydrophobic solution,which may be the mechanism of its antipruritic effect.2.It was successful that the method of bile duct ligation was used to make the rat model of cholestasis.Mahuang Lianqiao Chixiaodou Decoction can significantly reduce the cholestasis itch model rats spontaneous scratching frequency,suggest that the treatment of Mahuang Lianqiao Chixiaodou Decoction in the cholestasis itching model has certain effect;3.The occurrence of pruritus may be related to the expression of PAR2 and TRPV1 in rats with cholestasis.4.The mechanism of the treatment of Mahuang Lianqiao Chixiaodou Decoction in the cholestasis itching may be related to the regulation of TRPV1 expression. |
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