Study On Protective Effect Of Extracts Of The Chinese Medicine Shaoyao And Gancao To DRGn Damaged By LPS

Objective By establishing the cell model of dorsal root ganglion neurone(DRGn)which were impaired by LPS,to study whether the extracts of the Chinese medicine Shaoyao and Gancao have protective effects on SIRT1-mTOR pathway.Method We used LPS at the concentration of 100 ng/m on the DRGn of SD fetuses,then resected DRG by microsurgery.Cells were divided randomly into four groups,8 samples in each group:control group(normal lab rat serum effected by LPS for 24 hours),LPS group(DRGn of SD fetuses effected by LPS for 24 hours with 24 hours reaction of the normal lab rat serum),SGD group(DRGn of SD fetuses reacted with gancao and shaoyao serum for 24 hours),SGD+Anti-SIRT1 group(nicotinamide 98nM effected DRGn of SD fetuses reacted with LPS for 24 hours then reacted with gancao and shaoyao serum for 24 hours).In this study,the dose of LPS was 100ng/mL,and the dosage of SGD was 60mg/kg.At the same time,the prepared SGD rats were given intragastric administration according to the grouping and experimental requirements.The orbital blood samples were collected at room temperature and the supernatant was taken after 30min.The preparation of serum containing SGD was completed.The morphology of the DRGn was observed by microscope and the survival rate of DRGn was detected by CCK-8 test.The expression levels of SIRT1,mTOR,p-mTOR,Ac-NF-κB were detected by Western Blot method.Finally,we did statistical analysis on the experimental results.Results The results of activity of cells under microscope showed that the morphology of DRG neurons in LPS group was significantly higher than that in group Con,and the survival rate was lower(P<0.01).The activity of each group was detected by CCK-8.The results showed that the activity of dorsal root ganglion neurons in group SGD and group SGD+anti-sirt1 was significantly higher than that of group LPS(P<0.01,P<0.05).Compared with group SGD,the cell activity of group SGD+anti-sirt1 was significantly lower(P<0.05).The results of Western and Blot showed that the expression of SIRT1 and p-mTOR in LPS group was lower than that in group Con,and the expression of mTOR and Ac-NF-κB increased(P<0.05).In group SGD,the expression of SIRT1 and mTOR increased,and the expression of p-mTOR、Ac-NF-κB and NF kappa B decreased(P<0.05)compared with the pure LPS group.The expression of SIRT1 and mTOR in SGD+anti-sirtl group was lower than that in group SGD,and the expression of p-mTOR、Ac-NF-κB was increased(P<0.05).Conclusion First,gancao and shaoyao decoction can effectively reduce the apoptosis of dorsal root ganglion neurons damaged by the dorsal root ganglion neurons inflammation induced and activated by lipopolysaccharide,and increase the data of dorsal root ganglion neurons.Second,in the LPS induced inflammatory injury,SIRT1 expression decreased,and its mechanism may be related to the increasing expression of mTOR,Ac-NF-,NF,B.Third,gancao and shaoyao decoction can increase the expression of SIRT1,promote the transformation of mTOR to p-mTOR,and reduce the expression of Ac-NF-kappa B,that is,through the SIRT1-p-mTOR-Ac-NF-kappa B pathway to play its anti-inflammatory and analgesic effects.Subject words Modified Shaoyao Gancao Decoction,Lipopolysaccharides,dorsal root ganglion neurone,silent mating type information regulation 2 homolog 1 anti-tumor.