Mechanisms Of Lung Inflamm-aging And Intervention Study Of Resveratrol

ObjectivesTo screen the candidate inflamm-aging-related genes and the target genes interfered with Resveratrol (Res) by using gene chip technology. And to detect the expression levels of inflamm-aging-related inflammatory cytokines IL-1β,IL-6, IL-10 and TNF-a mRNA in aged rats Lung tissue. Furthermore to study the mechanisms of inflammatory cytokines network in the development of inflamm-aging.Methods1.30 healthy male SD rats were randomly divided into 3 groups(each with 10 rats): young group with 4 months rats (4m), aged group with 24 months rats(24m) with distilled water intragastric administration for 3 months from 21 months, group with 24 months rats treated with Res by intragastric administration (60mg/kg/d) for 3 months from 21 months(24m+Res). The gene expression profiles of rat hippocampus and lung tissues from rats in 4m,24m,24m+Res groups were analyzed by using cDNA microarray with 112 genes of inflammatory cytokines and receptors respectively.The differential expressed genes between 24m and 4m groups,24m+Res and 24m groups were screened respectively. The mRNA from rat hippocampus and lung tissues in every group was labeled respectively with cy-3 and cy-5 separately by RT-PCR as cDNA probes. Then two probes were mixed and hybridized with the above-mentioned gene chip. Scan Array 3000 was used to scan the hybridized signals and the data analyzed by bioinformatics method.2. Total RNA was extracted from lung tissue of rats. Real time PCR were employed to show different expression of inflamm-aging-related inflammatory cytokines IL-1β, IL-6, IL-10 and TNF-a mRNA among three groups.Results1. In rat lung tissues, compared with those in 4m group, there were 10 differentially up-regulated (SLR>2) expressed pro-inflammarory genes in 24m groups, which were Cxcl2, IL-1α, IL-1β, IL-2, IL-1rⅡ, IL-6, IL-7, TNF,TNFsf4 and TNFsfll (P <0.05). with 3 down-regulated (SLR<-2) expressed anti-inflammatory genes in 24m group, which were IL-10rα, TGF-α and TGF-β1 (P<0.05). Compared with those in 24m group respectively, there were 5 up-regulated (SLR>2) expressed anti-inflammatory genes in 24m+Res group, which were IL-10, IL-10rα, Ccl2, TGF-β3 and TGF-βrⅡ(P<0.05). with 4 down-regulated (SLR<-2) expressed pro-inflammatory genes in 24m+Res group, which were IL-1β, IL-6, TNF and TNFsf4 (P<0.05). Those differential expressed genes include interleukins, tumor necrosis factors, chemotatic factors, adhesion molecules, etc.2. Compared with 4m group, the expression levels of TNF-α, IL-6 and IL-1β mRNA were significantly upregulated (P<0.01), while the expression level of IL-10 mRNA in 24m grope was significantly downregulated (P<0.01) in 24m grope. Compared with 24m group, the expression levels of TNF-α, IL-6 and IL-1β mRNA were significantly downregulated (P<0.01), while the expression levels of IL-10 mRNA were significantly upregulated (P<0.01) in 24m+Res group. The Realtime PCR results were consistent with the previous microarray results.Conclusion1. There is upregulation of genes expressions of pro-inflammatory cytokines and receptors and downregulation of genes expressions of anti-inflammatory cytokines and receptors in aged rats, which may eventually contribute to disequilibrium of the network of pro- and anti-inflammatory cytokines and receptors and disequilibrium of response of pro-and anti-inflammatory system. This may be the reason of high proinflammatory state in inflamm-aging.2. Res may interfere in inflamm-aging through rebuilding the new equilibrium of the network of pro- and anti-inflammatory cytokines and receptors and equilibrium of response of pro- and anti-inflammatory system..