The Effects Of MicroRNA-155 On Atherosclerotic Immuno-inflammatory Responses

Background: Atherosclerosis is the pathological basis of many important vascular events.In the current society,people's living standards have increased year by year,medical and health care has continued to improve,so that the aging population increased,leading to the incidence of senile diseases increased year by year,one of the most notable is atherosclerosis.At present,the cardiovascular disease caused by atherosclerosis in the global morbidity and mortality of the disease in the first place.At present,cardiovascular disease caused by atherosclerosis is at the top of the global morbidity and mortality rate.Therefore,it is of great significance to explore the pathogenesis and pathogenesis of AS in depth.Atherosclerosis is a very complex pathogenesis,involving a variety of factors.There is growing evidence that the role of immune inflammatory response in the development of atherosclerosis and the development process can not be ignored.Toll-like receptors?TLRs?mainly mediate non-specific immune responses and inflammatory responses,in which TLR4 is most closely related to human atherosclerosis,and the My D88-dependent pathway in TLR4 is most important.Micro RNA?miRNA?is a group of about 22 nucleotides of non-coding RNA,with the regulation of cell proliferation,with the regulation of cell proliferation,differentiation of the role of participation in a variety of organs and tissues of physiological and pathological mechanisms,in heart-related diseases The occurrence of development also has a regulatory role.Previous studies have found that miR-155 is a multifunctional miRNA that has been found,and a number of studies have shown that increased expression of miRNA-155 can inhibit the production of Toll-like receptor ligands and cytokines to induce proteolytic enzymes and affect the expression of multiple inflammatory factors.The levels of monocyte chemoattractant protein-1?MCP-1?,transforming growth factor-?1?TGF-?1?and inflammatory factor interleukin-10?IL-10?were associated with atherosclerosis Disease is closely linked.Previous studies have found that My D88 is the target gene for miR-155,and miR-155 can play a regulatory role by inhibiting the translation of My D88.The effect of miR-155 on the expression of My D88 in the process of atherosclerosis remains to be studied.Objective:The aim of this study was to investigate whether miR-155 regulates TLR4 signaling pathway in AS by Medullary Differentiation Factor 88?My D88?,a target gene Toll receptor-related factor,which plays an anti-atherosclerotic effect through immuno-inflammatory regulation.Method:First,Establishment of ApoE^-/-mouse AS model,a total of 10 male ApoE^-/-mice of 4-6 weeks of age were selected and fed with high-fat diet for 20 weeks.The AS model was established.Male 4-5 week old male C57BL/6 mice fed with 10 normal feeds were used as normal control.The formation of atherosclerotic plaque in the aorta was measured by HE staining.Immunohistochemistry was used to detect the expression of My D88 protein in arterial atherosclerotic plaques,the levels of IL-10,TGF-?1 and MCP-1 in serum were detected by enzyme-linked immunosorbent assay?ELISA?.Second,in vitro culture of mouse macrophages,given ox-LDL stimulation to build foam cell model.Experiment grouping:?1?miR-155 mimic group: transfected miR-155 mimic + ox-LDL;?2?miR-155 mimic negative control group: transfected miR-155 mimic negative control + ox-LDL;?3?miR-155 inhibitor group: transfected with miR-155 inhibitor + ox-LDL;?4?miR-155 inhibitor negative control group:transfected miR-155 inhibitor negative control + ox-LDL;?5?My D88 si RNA group:transfected My D88 si RNA + ox-LDL;?6?My D88 si RNA negative control group:transfected My D88 si RNA + ox-LDL.The expression of My D88 molecule and protein was detected by Real-time PCR and Western blot respectively.The levels of interleukin-10,TNF and MCP in inflammatory cells were measured by enzyme-linked immunosorbent assay?ELISA?.Result:First,ApoE^-/-mice after high fat diet,arterial HE staining showed significant atherosclerotic plaque,immunohistochemical results showed that My D88 showed significant atherosclerotic plaque,the levels of IL-10,TGF-?1 and MCP-1 in the atherosclerotic mice were significantly higher than those in the control group by enzyme-linked immunosorbent assay?ELISA?;the arterial HE staining of the normal control group showed no obvious atherosclerotic plaque,Immunohistochemical results showed that My D88 was negative in the blood vessels.Second,ox-LDL stimulates macrophages.Compared with miR-155 mimic negative control group,the expression of My D88 did not change significantly in miR-155 mimic group;protein level,My D88 expression was significantly reduced;the levels of IL-10,TGF-?1 and MCP-1 in the supernatant were significantly decreased?P <0.05?.Mi R-155 inhibitor group compared with miR-155 inhibitor negative control group,My D88 expression of the level of no significant changes;and the levels of IL-10,TGF-?1 and MCP-1 in the supernatant were significantly increased?P <0.05?;The expression of IL-10,TGF-?1 and MCP-1 in My D88 si RNA group was significantly lower than that in My D88 group?P <0.05?.Conclusion: 1.MyD88 and inflammatory factors were significantly increased in atherosclerotic model mice;2.miR-155 can inhibit the expression of My D88 protein;3.Increased miR-155 can reduce the release of inflammatory cytokines IL-10,TGF-?1 and MCP-1,and vice versa.