Fructus Tritici Levis is the dry,friable and lean fruit of the gramineous plant Triticum aestivum L.It is the by product of the flotation and grading of raw wheat grains.It is widely used in the treatment of hyperhidrosis.The market survey found that the quality of Fructus Tritici Levis is uneven,and the antiperspirant mechanism off Fructus Tritici Levis is not clear.Therefore,this topic mainly focuses on the quality evaluation of Fructus Tritici Levis,protein bioinformatics analysis of Fructus Tritici Levis,evaluation and mechanism of Fructus Tritici Levis on antiperspirant effect in pilocarpine nitrate mice model,so as to clarify the pharmacology of Fructus Tritici Levis in the treatment of sweating.The study provides the theoretical basis for the industrial application of Fructus Tritici Levis.The specific experimental methods and results are as follows:1.Research content: Firstly,the components of 12 kinds of Fructus Tritici Levis from different producing areas were determined.The grey correlation model and HPLC fingerprint were established.to Evaluate the quality of Fructus Tritici Levis from different producing areas combined with chemometric method to screen the excellent gerpmlasm resources;secondly,Fructus Tritici Levis with the best comprehensive evaluation quality was analyzed by proteomics and bioinformatics.The molecular mechanism and function of Fructus Tritici Levis protein were discussed to provide reference for functional evaluation of Fructus Tritici Levis.;then establish a mouse model of pilocarpine sweat syndrome,and intervene by administering Fructus Tritici Levis.Compare the number of sweat spots,the morphology of sweat gland cells,and the number of vacuoles in different groups of mice to evaluate the antiperspirant effect of different doses of Fructus Tritici Levis;Finally,determine the physiological and biochemical indicators of serum and brain tissue,and the expression of acetylcholine receptor protein in brain tissue.Compare the differences between different groups of mice,and explore the antiperspirant mechanism of Fructus Tritici Levis.Compare the differences between different groups of mice to explore the antiperspirant mechanism of Fructus Tritici Levis.2.Experimental results:(1)Quality evaluation of Fructus Tritici Levis from different producing areas: Based on the grey correlation method to evaluate the quality of 12 kinds of Fructus Tritici Levis from different origins,the relative correlation of Fructus Tritici Levis from different origins is between 0.35 and 0.67.According to the relative correlation,the ranking is: Jinan,Shandong >Zhengzhou,Henan >Baoding,Hebei>Yuncheng,Shanxi>Shijiazhuang,Hebei>Xiayi,Henan>Mianyang,Sichuan>Guilin,Guangxi>Fuyang,Anhui >,Haozhou,Anhui>Hegang,Heilongjiang;Establish fingerprint maps of 12 different Fructus Tritici Levis yields by HPLC and obtain a common chromatogram.There are 16 peaks with similarity between 0.936 and 0.996,which can be grouped into 3 categories.The quality ranking is based on 16 common peaks: Jinan,Shandong>Xiayi,Henan>Yuncheng,Shanxi>Baoding,Hebei>Fuyang,Anhui>Guilin,Guangxi>Zhengzhou,Henan>Mianyang,Sichuan>Leshan,Sichuan>Shijiazhuang,Hebei>Haozhou,Anhui>Hegang,Heilongjiang;Establish two quality evaluation methods for Fructus Tritici Levis,Fructus Tritici Levis of Jinan Shandong ranked first in both quality evaluation systems,,so Jinan Shandong Fructus Tritici Levis was used in the following experimental research.(2)Proteomics and bioinformatics analysis of Fructus Tritici Levis: Through proteomic analysis of Fructus Tritici Levis,a total of 7391 peptides were obtained,7390 proteins and2542 proteomes were identified.Fructus Tritici Levis proteins are mainly composed of histone family,gluten,heat shock protein,and pectin acetylesterase family and uncharacterized proteins;bioinformatics analysis of Fructus Tritici Levis proteome,Fructus Tritici Levis protein involves 16 molecular functions,13 cell components and 20 biological processes at Level 2;KEGG metabolic pathway analysis,Fructus Tritici Levis protein is mainly Participate in protein processing in endoplasmic reticulum,Ribosome,Glycolysis / Gluconeogenesis and other 192 metabolic pathways;predict and analyze Fructus Tritici Levis protein interaction network,Fructus Tritici Levis protein is mainly involved in the three network processes of genetic information processing,biological information metabolism and energy metabolism.These results provided reference for pharmacodynamics analysis and functional evaluation of Fructus Tritici Levis.(3)Antiperspirant effect of Fructus Tritici Levis on pilocarpine nitrate sweat in mice: To explore the antiperspirant effect of Fructus Tritici Levis on pilocarpine nitrate sweat in mice.During the experiment,the mice were weighed seven times,and it was found that Fructus Tritici Levis did not interfere with the weight of mice(P>0.05);After 1 d,7 d and 14 d of the same dose,the antiperspiration effect of Fructus Tritici Levis was significant(P<0.05),while the effect of wheat was not(P<0.05).he medium and high doses of Fructus Tritici Levis had a significant antiperspirant effect on 1 d,7 d,and 14 d(P<0.05).The antiperspiration effect of low-dose Fructus Tritici Levis was observed only at 14 d after administration(P<0.05).He staining was performed on the soles of mice’s feet.After the injection of pilocarcinine nitrate,the sweat gland cells expanded significantly,and the incidence of subnuclear vacuoles in unit sweat gland increased significantly.medium and high doses of Fructus Tritici Levis can reduce the incidence of subnuclear vacuoles in mouse sweat gland cells(P<0.05).(4)Effect of Fructus Tritici Levis on physiological and biochemical indexes and Ach R receptor protein in mice with sweat syndrome: Analyze the effect of Fructus Tritici Levis on the physiological and biochemical indexes and Ach R receptor protein in sweating mice.After observing the sweat spots on the soles of the mice,the mice were dissected,and the serum and brain tissue were collected to measure the content of acetylcholine,(Ach)acetylcholinesterase(Ach E),calcium and noradrenaline(NE),and the expression of acetylcholine receptor(Ach R)in mouse brain tissue.The results showed that pilocarpine nitrate can increase the content of serum Ach and the expression of Ach R receptor in mouse brain tissue(P<0.01),but there was no significant effect on Ach E,calcium and NE(P>0.05);high-dose Fructus Tritici Levis can significantly reduce mouse serum Ach content(P<0.01);medium-dose and high-dose Fructus Tritici Levis can reduce Ach content in mouse brain tissue(P<0.05);medium and high-dose Fructus Tritici Levis can reduce Ach R expression in mouse brain tissue(P<0.01).The grey correlation evaluation method and HPLC fingerprint method can effectively evaluate the quality difference of Fructus Tritici Levis from different producing areas and screen high-quality gerpmlasm resources;Fructus Tritici Levis can effectively improve the symptoms of sweating in mice with pilocarpine nitrate,reduce the content of acetylcholine and the expression of acetylcholine receptor protein of brain tissue in mice.which further revealed the medicinal mechanism of Fructus Tritici Levis in the treatment of sweat syndrome. |