| ObjectiveWith qiongyugao intervention of D-galactose induced subacute aging model rats in the hypothalamus and the research site, on the basis of preliminary studies, the protein group of iTRAQ technology for mass spectrometric identification search qiongyugao intervention decline boss in hypothalamus of differential proteome analysis, find out the obvious differences in multiple candidate proteins. And then refer to the relevant delay the aging literature and biological information analysis technology of differentially expressed protein analysis, ultimately determine hypothalamic inflammation aging related proteins induced, and the conducted related experiments to verify the differences in protein expression levels, further inquiry of qiongyugao retard the aging mechanism of action in the hypothalamus, while guiding the hypothalamic inflammation caused by aging in the future.Methods1. The preliminary experiment by proteome of iTRAQ technology for mass spectrometric identification search qiongyugao intervention bad boss in hypothalamus of differential proteins were analyzed, identified by comparison qiongyugao group and model group of 295 species differences in protein screened fold difference is more than or equal to 1.30 proteins.2. The fold difference is more than or equal to 1.30 differential proteins were screened and sequence alignment, by referring to the differential proteins related to the aging literature, and biological information analysis, find qiongyugao delay the aging of the candidate target proteins in determining and hypothalamic inflammation pathway related proteins.3. The immunoblotting (Western blot and important index of fluorescence quantitative PCR (Q-PCR) analysis, observation of qiongyugao intervention hypothalamic inflammation associated protein expression level changes, through changes in hypothalamic inflammation related proteins with different expression levels explain qiongyugao to intervene inflammatory changes in the hypothalamus delaying aging mechanism, ultimately determine the hypothalamic inflammation caused by the change of the mechanism of aging. Results1. The pre experiment to find to qiongyugao group and model group of 295 species differences in protein analysis successfully screened out fold difference is more than or equal to 1.30 proteins.2. By screening out fold difference is more than or equal to 1.30 for the differences in protein and sequence alignment, consulting differential proteins related to aging literature, biological information analysis and successfully identified the hypothalamus inflammatory pathways related protein NF kappa B protein, pip5klc protein, PTPRC/CD45 protein.NF-κB protein activation is a typical manifestation of inflammatory reaction, through the detection of NF-κB protein activation can body reaction inflammation; pip5klc protein and PTPRC/CD45 protein in anti-inflammatory process has certain positive role and the expression of the two in a certain extent, reflect the anti-inflammatory ability.3. The NF-κB protein, pip5klc protein, PTPRC/CD45 protein for Western blot and Q-PCR, found Qiongyu extract inhibited the expression of NF-κB protein in the aging process, slowed the pip5klc protein and PTPRC/CD45 protein in aging process decreased.ConclusionQiongyugao stem down-regulation of inflammatory pathways associated protein is the NF kappa B protein, which can lead to inflammation of the hypothalamic inflammation activated protein; pip5klc protein and PTPRC/CD45 protein up regulation of inflammatory pathway related proteins, which can against the protein of inflammatory reaction and inflammatory related diseases. Through the experimental study concluded that the mechanism can more clearly explain for suppression and slow and that inhibition of pro-inflammatory proteins (such as NF kappa B protein) in the adverse external environment under the influence of the activation of slow anti-inflammatory proteins, such as pip5klc protein, PTPRC/CD45 protein under the adverse external environment influence the severity of the recession. Through bidirectional regulation and control of the related proteins of the hypothalamus inflammatory pathway to reduce the inflammation of the hypothalamus, avoid the decline of the physiological function of the body caused by the inflammation of the hypothalamus, and then achieve the purpose of delaying the aging of the body.Hypothalamic inflammation can cause aging and its mechanism with many inflammatory pathway related proteins have a certain relationship, hypothalamic inflammation in the future lead to aging experiment can select other inflammatory pathway related proteins were verified and can more fully explore hypothalamic inflammation caused by the mechanism of aging. |
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