| Objective:Preliminary clarify biological basis of syndrome between four kinds of benefit formula and different aging stages through comparative study on the variety of liver proteome of 3,6,12 month male mice and four kinds of benefit formula effect on the callback function of mice's differentially expressed proteins(DEP)of different ages,in order to provide a new theoretical basis and data support for the comprehensive analysis of aging mechanism and prevention measures.Method:10 ICR male mice of 3month were selected as control group.60 ICR male mice of 6 month were randomly divided into groups of Si Jun Zi decoction,Si Wu decoction,Liu Wei Di Huang decoction,Jin Gui Shen Qi decoction,vitamin E and the control group.Dividing the mice of 12 month with the same method.Each group contains 10 mouse.Drug groups was treated with intragastric administration while the young control group and control groups of 6 month and 12month intragastric the same volume ratio physiological saline.After administration mouse were sacrificed in the thirty-first days.Quick freezing with liquid nitrogen after taking out the liver,put it into 80 degrees below zero refrigerator as soon as possible.Extraction of liver mitochondria.Based on iTRAQ technology,joint LC-MS/MS mass spectrometry,screening of DEP through the UniProt release(2016-07)database.Analysis of DEP in the above groups through GO analysis and KEGG pathway analysis,intersection processing DEP,so as to further screening of protein markers.Result:Control group of 3 month and 6 month both have 358 DEP.Control group of 6 month and 12 month both have 347 DEP.Control group of 3 month and 12 month both have 365 DEP.Analysis of 6 month group,control group and Si Jun Zi decoction group both have 443 DEP,control group and Si Wu decoction group both have 412 DEP,control group and Liu Wei Di Huang decoction group both have 406 DEP,control group and Jin Gui Shen Qi decoction group both have 364 DEP,control group and Vitamin E group both have 370 DEP.Analysis of 12 month group,control group and Si Jun Zi decoction group both have 336 DEP,control group and Si Wu decoction group both have 340 DEP,control group and Liu Wei Di Huang decoction group both have 402 DEP,control group and Jin Gui Shen Qi decoction group both have 373 DEP,control group and Vitamin E group both have 338 DEP.After analyzing,a total of 97 aging-related protein markers were screened.There were 32 DEP which were regulated by Si Jun Zi decoction related to Qi deficiency syndrome.There were 36 DEP which were regulated by Si Wu decoction related to blood deficiency syndrome.There were 30 DEP which were regulated by Liu Wei Di Huang decoction related to Yin deficiency syndrome.There were 26 DEP which were regulated by Jin Gui Shen Qi decoction related to Yang deficiency syndrome.vitamin E affects 8 age related DEP.Through the GO analysis of 97 age-related DEP,we found 54 DEP were enriched in the 35 biological processes which were related to growth and development.Si Jun Zi decoction regulates 21 DEP related to growth and development,Si Wu decoction regulates 24 DEP related to growth and development,Liu Wei Di Huang decoction regulates 17 DEP related to growth and development,Jin Gui Shen Qi decoction regulates 11 DEP related to growth and development.97 DEP associated with aging were identified in this study,among them,we found a total of 54 DEP were enriched in 35 biological processes related to growth and development.The four benefit formula can promote the growth and development of the body through the regulation of DEP.Si Jun Zi decoction regulates 21 DEP which are related to the growth and development.Si Wu Decoction regulates 24 DEP which are related to the growth and development.Liu Wei Di Huang decoction regulates 17 DEP which are related to the growth and development.Jin Gui Shen Qi decoction regulates 11 DEP which are related to the growth and development.Each group of DEP were occurred simultaneously.24 DEP were regulated by both Si Jun Zi decoction and Si Wu decoction.9 DEP were regulated by both Liu Wei Di Huang decoction and Jin Gui Shen Qi decoction.3 DEP were regulated by both Jin Gui Shen Qi decoction and Si Jun Zi decoction.1 DEP were regulated by both Liu Wei Di Huang decoction and Si Wu decoction.Conclusion:This study find the protein markers related with aging on ICR male mice liver of 3,6,12 month by proteomics technology.On this basis,using four benefit formula as a probe,to explore the correlation between aging and imbalance of Qi,blood,Yin and Yang through analysis the effect of changing in proteome which is intervened by four benefit formula.Draw the following conclusions:1 In the aging process,the homeostasis of the organism is gradually unbalanced.In this study,we identified 97 DEP related to aging,which are related to the PI3K-Akt signaling pathway,fatty acid metabolism,peroxisome,cancer pathways,ECM-receptor interaction,PPAR signaling pathway,ubiquitin-proteasome system,insulin signaling pathway,mTOR signaling pathway,longevity regulating pathway.2 Si Jun Zi decoction,Si Wu decoction,Liu Wei Di Huang decoction and Jin Gui Shen Qi decoction could respectively regulate mouse proteome changes of aging process by enriching Qi,enriching blood,enriching Yin or enriching Yang.2.1 32 DEP which are regulated by Si Jun Zi decoction with the function of enriching Qi,mainly enriched in the PI3K-AKT signaling pathway,peroxisome,proteasome,longevity regulating pathway and ECM receptor interaction pathway,etc.Si Jun Zi decoction has improved the biological changes due to aging which are also associated with the enriching Qi.2.2 36 DEP which are regulated by Si Wu decoction with the function of enriching blood,mainly enriched in the PPAR signaling pathway,FOXO signaling pathway,PI3K-AKT signaling pathway,longevity regulating pathway,etc.Si Wu decoction has improved the biological changes due to age which are also associated with the enriching blood.2.3 30 DEP which are regulated by Liu Wei Di Huang decoction with the function of enriching Yin,mainly enriched in the PPAR signaling pathway,peroxisome,ECM-receptor interaction pathway,fat digestion absorption,etc.Liu Wei Di Huang decoction has improved the biological changes due to age which are also associated with the enriching Yin.2.4 26 DEP which are regulated by Jin Gui Shen Qi decoction with the function of enriching Yang,mainly enriched in the PPAR signaling pathway,peroxisome,biosynthesis of unsaturated fatty acids,etc.Liu Wei Di Huang decoction has improved the biological changes due to age which are also associated with the enriching Yang.3 Through this study,we found a total of 54 DEP were enriched in 35 biological processes related to growth and development.The four benefit formula can promote the growth and development of the body through the regulation of DEP.Si Jun Zi decoction regulates 21 DEP which are related to the growth and development.Si Wu Decoction regulates 24 DEP which are related to the growth and development.Liu Wei Di Huang decoction regulates 17 DEP which are related to the growth and development.Jin Gui Shen Qi.decoction regulates 11 DEP which are related to the growth and development.4 24 DEP are co regulated by both Si Jun Zi decoction and Si Wu decoction.9 DEP are co regulated by both Liu Wei Di Huang decoction and Jin Gui Shen Qi decoction.3 DEP are co regulated by both Jin Gui Shen Qi decoction and Si Jun Zi decoction.1 DEP are co regulated by both Liu Wei Di Huang decoction and Si Wu decoction.It is suggested that there is a certain internal relation between Yin,Yang,Qi and blood,which needs further study.5 It was found that vitamin E does not have a clear role in the regulation of protein level on the mouse of 6 month and 12 month through the intervention of vitamin E on the aging mice. |
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