Study On The Mechanism Of Lilac,Curcuma Aromatica And Kaki Calyx Decoction In Treating Acute CINV By Regulating 5-HT And 5-HT3 Receptor

Objective Replication the pica model in acute CINV rats by adopting the method of Chronic restraint and injection of Cisplatin,to explore the modern scientific and material foundation of the "liver depressing spleen,stomach Qi rising" syndrome of TCM from protein quantitative analysis and gene expression level.The lilac,curcuma aromatica and kaki calyx decoction,a new prescription with "soothing the liver,strengthening the spleen and stomach and suppressing superinverse" as the treatment method,was used to intervene in rats.By observing the symptoms of rats and detecting related indexes,to make clear the therapeutic effect of "soothing the liver,strengthening the spleen and stomach and suppressing superinverse" method on acute CINV.By observing the changes of the content of 5-HT in the rat serum and the ileum,and the expression of 5-HT3 R in the ileum and medullary tissues before and after the administration,to reveal the partial mechanism of "soothing the liver,strengthening the spleen and stomach and suppressing superinverse" method in the prevention and treatment of acute CINV.Methods A total of 48 male Wistar rats are adopted at the SPF level and adapted for one week in the laboratory.Replication the pica model in acute CINV rats by adopting the method of chronic restraint and injection of cisplatin.Then divide the experimental animal into blank,model,ondansetron hydrochloride,lilac and kaki calyx decoction,only curcuma aromatica and lilac,curcuma aromatica and kaki calyx decoction group with a random number method.The drug dosage of 7.3,4.9 and 12.2 g·kg-1·d-1 is given to the group of lilac and kaki calyx decoction,only curcuma aromatica and lilac,curcuma aromatica and kaki calyx decoction for gavage treatment.The drug dosage of 2.6g·kg-1·d-1 is given to the reference group of ondansetron hydrochloride for gavage treatment.Taking the blood from the venae cava inferior and organization from ileum and oblongata after successful modeling.Then,determining the content of 5-HT and 5-HIAA in serum and the content of 5-HT,5-HIAA,MAOA and TPH1 in the ileum by the method of Elisa.determining the expression of 5-HT3 R gene in the ileum and oblongata by the method of RTq PCR.Results 1.The assessment of successful model of the acute CINV:(1)Sugar water preference experiment: 23-24 h after establishing the acute CINV model,the sugar water preference of the model group is lower than that of the blank group.The difference among them is statistically significant(P<0.01).The sugar water preference in each treatment group is higher than that of the model group in addition to the only curcuma aromatica group.The difference among them is statistically significant(P<0.01,P<0.05)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01).(2)Monitoring of vomiting in rats by kaolin intake: 12 and 24 h after establishing the acute CINV model,the intake of kaolin of the model group is significantly higher than that of the blank group.The difference among them is statistically significant(P<0.01).2.The assessment of the lilac,curcuma aromatica and kaki calyx decoction efficacy: Monitoring of vomiting in rats by kaolin intake: 12 h after establishing the acute CINV model,the intake of kaolin in each treatment group is lower than that of the model group.The difference among them is statistically significant(P<0.01).12-24 h after establishing the acute CINV model,the intake of kaolin in each treatment group is lower than that of the model group in addition to the only curcuma aromatica group.The difference among them is statistically significant(P<0.01).0-24 h after establishing the acute CINV model,the intake of kaolin in each treatment ,group is lower than that of the model group.The difference among them is statistically significant(P<0.01,P<0.05)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01).3.Determining the content of 5-HT and 5-HIAA in serum and the content of 5-HT,5-HIAA,MAOA and TPH1 in the ileum in each group of rats by the method of Elisa,and calculating the ratio of 5-HIAA/5-HT content: The content of 5-HT in serum and the ileum in the model group is higher than that of the blank group.The difference among them is statistically significant(P<0.01).The content of 5-HT in serum in each treatment group is lower than that of the model group in addition to the only curcuma aromatica group.The difference of the lilac and kaki calyx decoction group is no statistically significant(P> 0.05).The difference of ondansetron hydrochloride and lilac,curcuma aromatica and kaki calyx decoction group is statistically significant(P<0.01).The content of 5-HT in serum and the ileum in each treatment group is lower than that of the model group in addition to the only curcuma aromatica group.The difference among them is statistically significant(P<0.01,P<0.05)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01).The content of 5-HIAA in serum and the ileum in the model group is lower than that of the blank group.The difference among them is statistically significant(P<0.01,P<0.05).The content of 5-HIAA in serum in each treatment group is higher than that of the model group in addition to the only curcuma aromatica group.The difference of lilac and kaki calyx decoction group is statistically significant(P<0.01).The content of 5-HIAA in the ileum in each treatment group is higher than that of the model group.The difference among them is statistically significant(P<0.01,P<0.05)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01). The ratio of 5-HIAA/5-HT content in serum and the ileum in the model group is lower than that of the blank group.The difference among them is statistically significant(P<0.01,P<0.05).The ratio of 5-HIAA/5-HT content in serum and the ileum in each treatment group is higher than that of the model group in addition to the only curcuma aromatica group.The difference among them is statistically significant(P<0.01)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01).The content of MAOA in the ileum in the model group is lower than that of the blank group.The difference among them is statistically significant(P<0.01).The content of MAOA in the ileum in each treatment group is higher than that of the model group.The difference of the only curcuma aromatica group is no statistically significant(P> 0.05).The difference of the ondansetron hydrochloride,lilac and kaki calyx decoction and lilac,curcuma aromatica and kaki calyx decoction group is statistically significant(P<0.01).The content of TPH1 in the ileum in the model group is higher than that of the blank group.The difference among them is statistically significant(P<0.01,P<0.05).The content of TPH1 in the ileum in each treatment group is lower than that of the model group in addition to the only curcuma aromatica group.The difference among them is statistically significant(P<0.01)and the most significant one is the lilac,curcuma aromatica and kaki calyx decoction group(P<0.01).4.determining the expression of 5-HT3 R m RNA in the ileum and oblongata by the method of RT-q PCR: The expression of 5-HT3 R m RNA in the ileum is higher than that of the blank group.The difference among them is statistically significant(P<0.01).The expression of 5-HT3 R m RNA in the ileum in each treatment group is lower than that of the model group in addition to the only curcuma aromatica group.The difference of the ondansetron hydrochloride and lilac and kaki calyx decoction group is no statistically significant(P> 0.05).There is a significant difference in the group of lilac,curcuma aromatica and kaki calyx decoction group and the difference is statistically significant(P<0.01).The expression of 5-HT3 R m RNA in the oblongata is higher than that of the blank group.The difference among them is statistically significant(P<0.01).The expression of 5-HT3 R m RNA in the oblongata in each treatment group is lower than that of the model group.The difference among them is statistically significant(P<0.01,P<0.05).Conclusion 1.The pica model of liver depressing spleen in acute CINV rats can be successfully replicated by Binding stress response in rats by using a binding device and injecting of cisplatin.2.The liver depressing spleen model in acute CINV is related to the protein expression of 5-HT and the gene expression of 5-HT3 R.Reducing the level of TPH1 in the ileum tissue,increasing the level of MAOA,inhibiting the synthesis of 5-HT,promoting the metabolism of 5-HT and reducing the expression of 5-HT3 R in the ileum and oblongata tissues is possible to be a new direction for the treatment of the liver depressing spleen model in acute CINV.3.In the treatment of the liver depressing spleen model in acute CINV,lilac with curcuma aromatica indeed has the function of reducing toxicity and increasing efficacy.