| Background: Throughout the ages,people have been seeking to explore the mechanism and try to slow the progress of individual aging.Under the current social situation with so many elderly people,more and more studies have shown that elderly individuals are more prone to various pathological changes and cause diseases.Among all diseases,cardiovascular disease is the first to be affected by aging.At the same time,many manifestations of cell aging during aging are also important factors in the development of cardiovascular pathological changes.At the same time,the cardiovascular system is a maintenance system that is particularly important for living individuals.Therefore,studying the aging mechanism of the cardiovascular system is currently a very valuable research.The intima is the core part of the normal blood vessel structure,and the endothelial cell is an extremely important cell among the many components involved in the construction of the intima of the blood vessel.Vascular endothelial cells perform functions including phagocytosing abnormal foreign and endogenous materials,bacteria and necrotic damaged tissue cells and aging tissue cells,and are also connected to a variety of functional activities of the body,which mainly include ensuring the integrity of blood vessels,controlling the passage of substances and Transport of leukocytes to and from the blood,blood clotting(thrombosis and fibrinolysis),inflammation,new blood vessel formation(angiogenesis),vasoconstriction and vasodilation,blood pressure control,etc.During the process of vascular aging,its original normal function slowly declines,resulting in increased wall wall hardness,thickened,abnormally reduced or enlarged lumen.Vascular abnormalities can affect disease occurrence and development in different ways.Cell aging is becoming a key mechanism for the attenuation of vascular endothelial activity during aging.However,the molecular mechanisms that cause adverse changes in vascular endothelial function during aging are not fully understood.Since the last century,groups of researchers have continued to explore related cells.Everyone has a more comprehensive and profound understanding of helper T cells,and has discovered a different subset of traditional Th1 and Th2 and Treg.Finally,Th17 cells were defined as a separate subset of cells in the Th population.Th17 cells secrete factors related to individual activities and affect all aspects of individual life activities.Its cell name was also obtained because it significantly produced IL-17(mainly IL-17A).Th17 cells are responsible for triggering inflammatory damage and are polarized at the site of inflammation under the joint or independent action of multiple pro-inflammatory cytokines.IL-17 is not only associated with inflammation in the first place,but also induces inflammatory responses by inducing the expression of other related factors,leading to cell dysfunction.Th17 is a type of cell that plays a crucial role in normal conditions and disease states.It participates in all aspects of an individual's immune response,especially inducing inflammatory processes.In aging,chronic inflammation is often inevitable.At present,some scholars have proposed the relationship between aging and inflammation.It is believed that it is the inflammation that occurs in the aging process,which stimulates the body to react variously,such as oxidative stress,and thus becomes one of the important reasons for the aging process.However,the molecular mechanisms underlying Th17 and endothelial aging have not been reported.Objective: To investigate the effect and significance of Th17/IL-17 in the aging of mouse aortic endothelial cells,and to analyze the possible mechanism of its effect on the senescence of endothelial cells,in order to provide new ideas for the diagnosis and treatment of aging-related cardiovascular diseases.Methods: In this study,the aortic endothelial cells of healthy C57 BL / 6J mice were used as the research object.First,the proportion of Th17 cells in the spleen cell suspension of young middle-aged and elderly mice was detected by flow cytometry,and each group was detected by Western blot.Expression of senescence-related proteins in endothelial cells.Then the mouse aortic endothelial cell line was used as the research object for in vitro intervention.The endothelial cells were treated with IL-17 A at different concentrations to observe changes in cell morphology and related inflammation indicators.Cell senescence was observed by ?-galactosidase staining.After determining the optimal concentration of IL-17 A,the cell cycle changes of IL-17 receptor A and one of the senescence manifestations of mouse aortic endothelial cells were detected by flow cytometry.Western blot was used to detect the changes of senescence pathway protein expression and then to explore the mechanism of IL-17 A affecting senescence through the combined intervention experiments of specific pathway inhibitors.Results:(1)During ageing of C57 / B6 mice,the proportion of Th17 in the spleen cell suspension gradually increased,and the expression rate of Th17 cells(2.61 ± 0.34%)in the spleen stimulation of the old rats was significantly higher than that in healthy young rats(0.79 ± 0.17%)and healthy middle-aged rats(1.31 ± 0.21%)(P <0.01 compared with young and middle-aged),the expression rate of Th17 cells(37.52 ± 3.41%)was significantly higher after spleen stimulation in aged rats after induction Healthy young rats(20.88 ± 1.46%)and healthy middle-aged rats(26.00 ± 1.39%)(P <0.01 compared with young and middle-aged)and aortic endothelial cells expressed p53,p21,p19,Rb,and p16 Showing an increasing trend.(2)After IL-17 A acted on mouse aortic endothelial cells,the cells became senescent,and the positive rate of ?-galactosidase staining increased significantly.The expression of IL-17 receptor A on the cell membrane surface increased significantly,and cell cycle monitoring It was found that the G0 / G1 phase ratio increased significantly.(P <0.05)(3)After IL-17 A intervention,the expressions of cell senescence-related pathway proteins p53,p21,p19,Rb,and p16 were significantly increased(all P <0.01),and the upstream pathway NF-?B p65 protein,p38 MAPK protein,and JNK protein The level of phosphorylation increased significantly(all P <0.01),while the expression of ERK protein and its phosphorylation level did not change significantly.(4)After the IL-17 A intervention group was added with NF-?B pathway inhibitor PDTC,p38 MAPK pathway inhibitor SB203580,and JNK pathway inhibitor SP600125,the expression levels of senescence-related pathway proteins were significantly lower than the former(both P <0.01).Conclusion: Th17/IL-17 can induce the senescence of mouse aortic endothelial cells.The mechanism may be that IL-17 binds to IL-17 receptor A on endothelial cell membrane,thereby activating intracellular NF-?B,p38 and JNK pathways,and Furthermore,the p53 / Rb senescence pathway was activated to induce senescence of endothelial cells. |
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