| Background Aging is an inevitable physiological change that occurs with the passage of time.Aging is the most important risk factor for most non-communicable diseases(including cardiovascular diseases,cancer,diabetes and neurological diseases).In the elderly population,the physiological functions of many organs decrease with age.As the nutrient absorption organ with the largest surface area in the human digestive tract,the intestinal tract’s physiological function is severely affected by the aging process.After aging,intestinal tissue will also have a series of changes such as intestinal smooth muscle contraction ability,PH value of digestive juice and imbalance of intestinal flora.According to the theory of aging free radicals,free radicals and reactive oxygen species(ROS)produced by organisms during aging can cause continuous damage to biological macromolecules including nucleic acids,proteins and lipids.A large number of studies have shown that an increase in the level of oxidative stress will lead to an increase in the incidence of chronic inflammation.Therefore,detecting the expression of oxidative stress and inflammation-related cytokines in body organs can reflect the aging state of organ tissues.In this study,male C57BL6 mice were used as the research object to establish a naturally aging mouse model.The changes of oxidative stress levels and changes of inflammation-related cytokines in the intestinal tissue of mice during aging were examined.The roles of oxidative stress and chronic inflammation in the aging process were discussed.And the specific biomarkers that can judge intestinal tissue aging were preliminary screened.Materials and MethodsIn this study,male C57BL6 mice aged 4,5,8,and 20 months were selected as natural aging models.Immunohistochemical methods were used to detect changes in the level of nucleic acid oxidation in the intestinal tissue,and changes in the expression levels of anti-nucleic acid oxidation-related proteins MTH2,AUF1,OGG1,YB1 and other proteins.The protein chip method was used to detect the age-related changes of inflammation-related cytokines in mouse serum and intestine.Screening for inflammatory-related cytokines that increase in age in both serum and intestinal tissues.The selected adiponectin was verified by immunohistochemical method,and immunohistochemical staining of adiponectin was carried out in the five organs of intestine,spleen,liver,kidney and lung.ResultsThe results of immunohistochemistry showed that from 4,5,8 to 20 months in aging process,the expression levels of nucleic acid oxidation products 8-oxo G and 8-oxod G in the intestinal tissue have been increasing with ageing.In intestinal tissues,the expression levels of anti-nucleic acid oxidation-related proteins MTH2,AUF1,OGG1,and YB1 proteins also increased with age.The results of the protein chip showed that the expression level of adiponectin increased in both serum and intestinal tissues in 20-month-old mice compared with 4-month-old mice.Using immunohistochemistry,a confirmatory experiment was conducted on five organs of mice,and the results showed that the expression level of adiponectin only increased in the intestinal tissues with age,and did not change in the spleen,liver,kidney and lung.Conclusions1)The expression levels of nucleic acid oxidation products 8-oxo G and 8-oxod G in mouse intestinal tissues have been increasing with age,suggesting that the level of oxidative stress in intestinal tissues increases with the aging of mice.2)The level of anti-nucleic acid oxidized protein in the intestinal tissue increases with age,indicating that the intestinal tissue has been improving its own antioxidant capacity to cope with the increase in oxidative stress.3)The expression level of adiponectin only increases with age in the serum and intestinal tissue of mice,but it does not exist in other organs,suggesting that adiponectin may be used as the intestine of mice.Specific markers of tissue aging. |
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