Exploration On Specific Proteins Of Kidney-Yin Deficiency Syndrome Based On The Correspondence Of Prescription And Syndrome

[Background]Kidney-yin deficiency syndrome is one of the basic syndrome of Traditional Chinese Medicine, the chinese ancient doctors paid attention to its syndrome differentiation and treatment. People started to experimentally study on the essence of kidney-yin deficiency syndrome in late 1950s. Especially in recent two decades, many scholars in enzymes, cell biology, immunology, molecular biology and other subject, found kidney-yin deficiency syndrome concerned with the hemorheology, endocrine, immune, material and energy metabolism, trace elements, biochemistry, enzymes and abnormal gene expression, and so on. They confirmed its preliminarymodern basis of the pathophysiology, and have made certain achievements.So far, there is no verdict for the research on essence of kidney-yin deficiency syndrome. It is well-known that proteins are the final expression forms of genomic and the primary executor of biological function. Hence, based upon the correspondence of prescription and syndrome, the project uses proteomics to study kidney-yin deficiency syndrome, thus creates a new method for the research on essentials of kidney-yin deficiency syndrome.[Objective]To find specific proteins of kidney-yin deficiency syndrome based upon the correspondence of prescription and syndrome. To explore the essence of kidney-yin deficiency syndrome on the expression level of protein. Furthermore, the occurrence mechanism of kidney-yin deficiency symptoms is explained.[Method]SD rats were randomly divided into three groups as follows:normal group, kidney-yin deficiency model group and Zuoguiyin group. kidney-yin deficiency model group:the rats were gavaged with thyroid tablets suspension to make model; Zuoguiyin group:according to the same method to make model, after the success of the model, Zuoguiyin were gavaged to rats; normal group:the same volume of water.After Zuoguiyin treatment, during anaesthesia of rats, to remove the liver, kidney, and serum.With 2-D electrophoresis technology to analyze and compare the expressing proteins in the liver, kidney, and serum of the three groups, and find the deferent proteins, i.e. the specific proteins of kidney-yin deficiency symptoms. Then these deferent proteins were determined by mass spectrographic analysis. Finally the determined deferent proteins of Kidney yin deficiency syndrome are and confirmed by Western-blot. According to the function of the specific proteins, the occurrence mechanism of clinical of kidney-yin deficiency symptoms is explained.[Result]1. Through Image Master 2D Platinum 6.0 analysis, under the same condition, the numbers of protein expression in deferent tissue two-dimensional gel electrophoresis profiles show:liver (1473±32), kidney (1351±28), serum (318±34).Use 3 times expression as a standard, we identify 21 differentially expressed protein spots, which Zuoguiyin regulate and correct in the model rats with kidney-yin deficiency symptoms.2.21 differentially expressed protein spots were detected by MALDI-TOP mass spectra, and 18 proteins spots obtain correct identification results. Among these proteins, the proteins which were highly expressed in model rats with kidney-yin deficiency, but were corrected by Zuoguiyi, include tyrosyl-DNA phosphodiesterase, Enol protein, tyrosine protein kinase 6, protocadherin, calneuron 1; the proteins which were highly expressed in model rats with kidney-yin deficiency, but were corrected by Zuoguiyi, include carbamoyl-phosphate synthetase 1, peroxiredoxin 1,signal recognition particle 9, fructose-bisphosphate aldolase A. The function of the different proteins was related with substance metabolism, signal transduction, anti-oxidation, etc.3. Aldolase A were and confirmed by western-blot. The map of western-blot shows:Aldolase A is expressed in the 3 groups, the expression level of is lowest in kidney-yin deficiency model group. This result is in full accord with gel analysis and mass identification.[Conclusions]1. The project screen and identify 18 specific proteins of kidney-yin deficiency symptoms. Among these proteins, the proteins which were highly expressed in model rats with kidney-yin deficiency, but were corrected by Zuoguiyi, include tyrosyl-DNA phosphodiesterase, Enol protein, tyrosine protein kinase 6, protocadherin, calneuron 1; the proteins which were highly expressed in model rats with kidney-yin deficiency, but were corrected by Zuoguiyi, include carbamoyl-phosphate synthetase l,peroxiredoxin 1, signal recognition particle 9,fructose-bisphosphate aldolase A. The function of the different proteins was related with substance metabolism, signal transduction, anti-oxidation, etc. We preliminarily explore occurrence mechanism of kidney-yin deficiency symptoms on the expression levels of protein.2. The result of western-blot is in full accord with gel analysis and mass identification.3. According to the correspondence of prescription and syndrome, it is feasible that the proteomics research on the essence of kidney-yin deficiency symptoms.