Serum Proteomic Analysis Of Rats Model Of Abnormal Phlegmatic Syndrome And Its Drug Intervention

Objective:To screening serum differential expression protein of rats model of abnormal Phlegmatic syndrome,to explore Candidate protein markers of abnormal Phlegmatic syndrome,clear drug targets.Methods:Selected SD rats of 90 mature male,take out of ten for the controlgroup?N?,the other in the model.To established animal models of abnormal Phlegmatic syndrome by spinach and coriander method into the cold humid environment.Weekly Watch biological characterization of rat,confirm Syndrome model.Experimental evaluation by APO erectile function experiment in rats selected impotence model.The impotence rats were randomly divided into modelgroup?A₁?and drug intervention group?A₃?.The non-impotence rats were randomly divided into modelgroup?B₁?and drug intervention group?B₃?.After two weeks,while observing biological characterization of the rats and evaluating erectile function.Using iTRAQ-LC-MS-MS technology to screen differentially expressed protein,to analyse the of biological information of this differentially expressed protein and to verify part of the candidate proteins by ELISA.Results:?1?At the period of model,the change in the model consistent with the generalstate of Uygur medicine general characteristics of abnormal Phlegmatic,that is curled upto less,liking to get together,slow,dull hair,chaos,clear urine,pale stool the soft stool,darktongue,greasy tongue coating,slow weight gain,increased food intake,decreased water intake,urine and stool output increased.After treatment withYimusake,the symptoms were impoved.?2?At the period of model,erectile latency of modelgroup was increased and the number of erections was reduced.The incubation periods of straddle,insertion and ejaculation were longer than normalgroup,and the frequency of straddle.insertion and ejaculation is lesser than normalgroup.After treatment with Yimusake,the incubation periods of straddle,insertion and ejaculation of B₃ were shorter than B_l;and the frequency of straddle,insertion and ejaculation of B₃ were more than B_l.?3?The iTRAQ results showed that:Serum identified a total of 318 protein,B₁ group were screened out 76 differentially expressed proteins,51 up-regulated,25down-regulated;B₃ group were screened from 77 differentially expressed proteins,57up-regulated,20 down-regulated;according to the biological information analysis and the combination of literature research to determine the B₁ group candidate mark protein 33,25 up-regulated,8 down-regulated;B₃ group candidate marker protein 23,21 up-regulated,2 down-regulated.?4?The ELISA results showed that:concentration of CRP,?2GP1 in B₁group was significantly higher than N group?P<0.05?,B₃ group and N group no significant difference?P>0.05?,B₃ group was significantly lower than B₁ group?P<0.05?;?1AGP,RBP4,AGT concentrations in B₁,B₃ group were significantly higher than N group?P<0.05?,B₃ group was significantly lower than B₁ group?P<0.05?;IGF1concentrations in B₁,B₃ group were significantly lower than N group?P<0.05?,B₃ group was significantly higher than B₁ group?P<0.05?.Conclusion:?1?The change in the abnormal phlegmatic syndrome and abnormal phlegmatic syndrome with impotence disease rats model consistent with the general state of Uygur medicine general charactersistics of abnormal phlegmatic,visible at the same time prolonged with the modeling factors influence,sexual function were reduced,sexual behavior were decreased and further confirmed the the model reliability and veracity,stability.?2?Through this study,preliminary identified 33 candidate serum proteins of abnormal phlegmatic syndrome rat model and 23 drug target candidate differentially expressed proteins.?3?Through the verification in animal serum,CRP,?2GP1,?1AGP,RBP4,AGT,IGF1,GPX1 were serum candidate proteins markers of abnormal phlegmatic syndrome,Up-regulation of CRP,?2GP1,?1AGP,RBP4,AGT;down-regulation of IGF1,GPX1 may plays a key role in the occurrence and development of abnormal phlegmatic syndrome,may become abnormal phlegmatic syndrome disease candidate protein markers,but still need to be further studied in sera of clinical patients.?4?On the occurrence and development of abnormal phlegmatic syndrome,chronic inflammation and metabolic dysfunction may be the core mechanism,but on the basis of clinical validation of the candidate proteins,further studies on the functional network of candidate protein markers will further reveal the role of chronic inflammation and metabolic function in the development of the syndrome.?5?Yimusake tablet is classic Uygur Medicine for the treatment of abnormal phlegmatic syndrome with impotence,premature ejaculation and oligoasthenospermia,it for treatment of abnormal phlegmatic syndrome disease may be the many targets,CRP,?2GP1,RBP4,?1AGP,AGT,GPX1,IGF1 were not only candidate protein markers of the syndromes,also may be the as Yimusake tablet treatment with abnormal phlegmatic syndrome disease targets.