To Study Of Chronic Stress On Neuropeptide Y And Receptor Y1 In Arcuate Of Rat Models Of Syndrome Of Liver-depression And Spleen-deficiency.

Objective:To observe the Changes of protein and gene expression of chronic stress on Neuropeptide Y and receptor Y1 in Arcuate nucleus of rat models of syndrome of liver-depression and spleen-deficiency.With this asa starting point to explore terminal with liver-depression and spleen-deficiency syndrome appeared nervous-endocrine basis of emotional estrangement, meanwhile to the effect of Xiaoyaosan(Taiping Huimin and agentof Bureau) as intervention drug use, revealing the mechanism of relieving the depressed liver and strengthening the spleen.Methods:This study established rat models of syndrome of liver-depression and spleen-deficiency with method of chronic immobilization stress.133 SD rats were randomly divided into five groups.Rats in the control groupwere bred physiological saline without stress.Rats in 7-days- the model group were bred physiological saline and adopted with stress 7 days.Rats in 21-days- the model group were bred physiological saline and adopted with stress 21 days.Rats in 7-days- the model group were bred physiological saline and adopted with stress 7 days,to establish models.Rats in the 7-days-Xiaoyaosan group were continuously stressed by method ofimmobilization and intragastric administration with Xiaoyaosan(3.854g/kg/day,70 kg per adult) for 7 days.Day 7 in the 8th day of the experiment,normal group,on day 21 in the 22 nd day of the experiment subjects.Toobserve changes of weight,food intake,drinking water,external appearanceand signs and to record those of rats in five groups.Toshow that could successfully reproduce the syndrome of liver-depressionand spleen-deficiency of rat model.Methods of immunofluorescence and RT-q PCR method were adopted to detect the expression of NPY and receptor Y1 in ARC.Results:Based on successful model,body weight and ingestion quantity of rats in the model group lower than control group,outward appearances make spirit are worse,unresponsive,yellow dark fur,ear claws luster fades,wet and dry stool is not adjusted,decreased mobility,while weightand ingestion quantity of rats in Xiaoyaosan group after three weeks better than the model group(P<0.05 或 P<0.01).NPY neurons and receptor Y1 weremarked with red fluorescence and distribute extensively in ARC;With macroscopic observation, the expression of receptor Y1 and NPY decreased at the end of 7 days and 21 days in model groups,however,it was opposite in the control group and the Xiaoyaosan groups.By RT-q PCR method,compared with the control group,expression of NPY m RNA and itsreceptor Y1 m RNA in hypothalamic ARC of rats in model group at theend of 7 days and 21 days,lower than the control group(P<0.05 或 P<0.01);compared with the model group,the expression of NPY m RNA and receptor Y1 m RNA in hypothalamic ARC of rats in Xiaoyaosan group,higher than the model group(P<0.05 或 P<0.01).Conclusions:Expression of NPY and receptor Y1 in ARC of model rat with syndrome of liver-depression and spleen-deficiency were with one accord.The study showed that NPY expression decreased and receptor Y1 expression increased in ARC of hypothalamus,could be to provide the experimental basis as one of the internal biological foundation on the syndrome of liver-depression and spleen-deficiency with manifestations of appetide decreasing,meanwhile to illustrate the target and mechanism of Xiaoyaosan on relieving the depressed liver,and invigorating the spleen-forregulating appetide.