Proteomics Analysis Based On ITRAQ Technology Of Acupuncture On The Hypothalamus Of Diet-induced Obesity Rats

Objective: To study the hypothalamus of diet-induced obesity rats by using proteomics technology to reveal the effect of obesity on organism and the weight-reducing effect of acupuncture intervention on diet-induced obesity rats and the related molecular mechanism,and to provide new ideas and targets for acupuncture treatment of obesity.Methods: The male SD rats were fed with high-fat diet,and the model of diet-induced obesity rats was established.Randomly selected 20 of the rats were divided into cure group(n=10),model group(n=10)and 10 normal feed rats as normal group.Acupuncture treatment on rats in cure group,taking acupoint: Zusanli,Zhongwan,Tianshu,Fenglong,Yinlingquan,Guanyuan,one time a day,last 30 minutes each time,a total of 28 days of treatment.The remaining two groups don't do any special treatment.The weight of rats was regularly tested and recorded,and then analyzed with SPSS22.0.After the intervention,the hypothalamus of three groups rats were taken out.The proteins of the hypothalamus were marked by iTRAQ,and separated and identified by LC-MS/MS.The differential proteins were analysised for further biological functional.Results :1.The weight changes of rats in the three groups were as follows:(1)After the high-fat diet model was established,the weight of the model group and the cure group was significantly higher than that of the normal group(P<0.01).(2)After acupuncture intervention,the weight of rats in the cure group was significantly lower than that before treatment(P<0.01).(3)After acupuncture intervention,the weight of rats in the cure group was significantly lower than that of the model group(P< 0.01).2.Separate and identify differential proteins of hypothalamus by LC-MS/MS:(1)3043 differantial proteins were identified;compared with the model group and normal group,19 proteins were down expressed and 34 proteins were were over expressed;compared with the cure group and model group,34 proteins were down expressed and 20 proteins were over expressed.(2)10 proteins were over expressed(or down expressed)in the model group and down expressed(or over expressed)in the cure group.3.GO function enrichment analysis :(1)In model group vs normal group,these over expressed identified proteins were mainly classified into structural constituent of myelin sheath,glyceraldehyde-3-phosphate dehydrogenase(NAD+)(phosphorylating)activity and oxidoreductase activity;and down expressed identified proteins were mainly classified into carboxylic acid binding and magnesium ion binding.(2)In cure group vs model group,these over expressed identified proteins were mainly classified into structural molecule activity and protein dimerization activity;and down expressed identified proteins were mainly classified in calmodulin-dependent protein kinase activity,protein serine/threonine kinase activity,glutamate receptor binding,endopeptidase activity,GTPase activity and ion binding.Conclusions:(1)High-fat diet may cause disorders of glucose and lipid metabolism,fat accumulation and obesity,and are closely related to the development of type 2 diabetes,cancer,and atherosclerosis.(2)The acupuncture program based on the theory of “invigorating spleen,removing dampness and warming kidney-yang”can significantly reduce the weight of obese rats and can be used in clinical practice.(3)Acupuncture treatment of obesity is the overall regulation of the body from multiple systems and multiple pathways.Its possible mechanism might be:(1)By inhibiting the activity of GAPDH or reducing its over-expression to regulates glucose metabolism and relieves cell apoptosis.(2)By inhibiting the activity of Ppp1r1b+ to inhibites food intake;or by promoting the dephosphorylation of PPP1R1 B to restores the activity of PP1 and regulates the glycogen metabolism.(3)By adjusting serum albumin levels up or down to adjusts glucose and lipid metabolism disorders.(4)By reducing the excessively high levels of GFAP to restore the structure and function of damaged nerve cells.(5)By inhibiting the over-expression of integrin ?1 to reduce the risk of atherosclerosis and cancer.