Study On The Effect Of Pilose Antler Peptide On "Zhuang Shen Yang,Qiang Jin Gu"

Objective: Objective to clarify the pharmacodynamic material basis of "Zhuang Shen Yang" of pilose antler,screen the active components of "Qiang Jin Gu" of pilose antler,determine the best process of enzymatic hydrolysis of polypeptide,and optimize the best preparation process of " Qiang Jin Gu " polypeptide enteric coated tablets of pilose antler.Method:1.Firstly,the different components of pilose antler were obtained by ultrafiltration and centrifugation,and the testosterone secretion of Leydig cells was used as the evaluation index for activity screening;the first-order mass spectrometry fingerprints of different batches of fresh and processed pilose antler were drawn by ESI-MS / MS,and the testosterone secretion of rat Leydig cells increased by each sample was determined.The first-order mass spectrometry data were analyzed by PCA,PLS-DA and opls-da Analysis: to realize the rapid differentiation of fresh and traditional processed velvet samples.Pls and gra were used to screen the active components with potential to promote the secretion of testosterone by Leydig cells,and the structure and composition of the possible active components were determined by secondary mass spectrometry data analysis.2.The effect of different components of Antler on MC3T3-E1 proliferation rate was determined by CCK8,and the active components of " Qiang Jin Gu " of antler were screened;the optimal enzymatic hydrolysis conditions of velvet antler > 10 k Da protein were optimized by response surface design;the granulation and tablet pressing process of the core were optimized by single factor investigation and orthogonal design.Finally,the velvet antler was prepared by controlling the speed of coating pot and the amount of coating solution Peptide enteric coated tablets.Result:1.The < 1 k Da component of pilose antler,i.e.oligopeptide,is the main active part of "Strengthening Kidney Yang";the components of fresh and processed pilose antler were significantly different,and both of them had good activity of promoting testosterone secretion by Leydig cells,and the more active components were Tyr-Ser-Phe(m/z 416)and Phe-Phe-Leu/Ile(m/z 426).2.The 3 ～ 10 k Da and 1 ～ 3 k Da components of pilose antler,i.e.the polypeptides of pilose antler,are the main "Qiang Jin Gu" active parts of pilose antler.The optimal enzymatic hydrolysis conditions of pilose antler > 10 k Da protein are as follows: temperature 50 ℃,time 4.25 h,p H7.7,enzyme to substrate ratio 6%.Under the optimal conditions,the yield of polypeptides of pilose antler is 19.73%.The results showed that the hardness of the chip core was suitable and non stick punching,the fluidity was good and the disintegration time was the shortest.By controlling the speed of coating pot 40 r / min and the dosage of coating solution8%,the enteric coated tablets of pilose antler polypeptide were prepared.Conclusion: The spectrum effect relationship between pilose antler and testosterone secretion of Leydig cells was studied,and the pharmacodynamic material basis of "Zhuang Shen Yang " of pilose antler was clarified.In addition,the active component of "Qiang Jin Gu" of pilose antler was identified as polypeptide,and the enzymatic hydrolysis process of "Qiang Jin Gu" polypeptide from pilose antler protein was optimized It provides a reference for the comprehensive utilization and development of Tricholoma matsutake.