| Coronary artery thrombosis is the main pathological basis of cardiovascular events in coronary heart disease.Before atherosclerotic thrombosis,the body will be in a hypercoagulable state conducive to thrombosis,that is,prethrombotic state.The analysis of the risk factors related to the prethrombotic state of coronary heart disease and the molecular mechanism of the prethrombotic state of coronary heart disease can provide a useful tool for the identification and diagnosis of the prethrombotic state of coronary heart disease.It can effectively interfere with the prethrombotic state of coronary heart disease and have important clinical significance for preventing the occurrence of cardiovascular events in coronary heart disease.The prethrombotic state of coronary heart disease is the result of abnormal changes of platelet,microcirculation,inflammatory reaction,coagulation state and fibrinolytic system.Inflammatory reaction is the key pathological link inducing coronary artery thrombosis.CANTOS clinical trials provide a clear clinical basis for the role of inflammation in atherosclerosis,and cardiovascular disease has entered a new era of targeting treatment with inflammation.Therefore,exploring the prethrombotic state from inflammatory reaction is of great significance in preventing coronary heart disease and cardiovascular events.MicroRNA?miRNA?is a kind of endogenous and highly conserved single strand non coding small RNA.It plays an important role in various major biological processes by degrading and inhibiting target genes and affecting protein translation.MiRNA can be detected at different stages of coronary heart disease and participate in the development and development of coronary heart disease.Therefore,screening the target miRNA related to the prethrombotic state of coronary heart disease and exploring its role in PTS can provide experimental basis and molecular targeting for preventing thrombus formation in prethrombotic state of coronary heart disease.Zedoarondiol is an effective component of Chinese medicine rhizoma curcumae,which is a half cup and terpenoid compound.Previous studies have reported that Zedoarondiol has the effect of inhibiting the inflammatory response,but it is not clear whether the mechanism of its anti-inflammatory action is related to the regulation of miRNA related to the prethrombotic state of coronary heart disease.Around the above,the subject has been studied in two aspects:?1?clinical analysis of risk factors related to pre thrombotic state of coronary heart disease;?2?the effect and role of Zedoarondiol intervention on RAW264.7 macrophage inflammation model by regulaing targets miRNA related to prethrombotic state.Part 1 Analysis of risk factors related to prethrombotic state of coronary heart diseaseObjective:To analyze the risk factors and clinical characteristics of prethrombotic state in patients with stable angina pectoris,and to explore the risk factors associated with prethrombotic state.Methods:115 patients with stable angina pectoris who underwent coronary angiography and the degree of coronary artery stenosis was more than 50%were selected.Through the Nailfold Microcirculation and platelet aggregation rate screening,we included 50 patients with stable angina pectoris and 65 patients without prethrombotic state.The general clinical data collection and laboratory examination were conducted to analyze the risk factors and clinical characteristics of prethrombotic state of coronary heart disease.Results:Compared with the clinical data of nonthrombus prethrombotic state group,the level of TC,TG and hs-CRP in prethrombotic state group of coronary heart disease was higher than that of nonthrombus prethrombotic state group,the difference was statistically significant?P?0.01?.Regression analysis showed that the levels of TC and hs-CRP were significantly correlated with prethrombotic state?P?<0.01?.The results of bivariate correlation analysis of platelet aggregation rate and clinical parameters showed that TC,TG,LDL-C and hs-CRP had significant correlation with platelet aggregation?P ? 0.01?.The subgroup analysis of prethrombotic state of coronary heart disease showed a significant correlation between platelet aggregation rate and hs-CRP level in prethrombotic state of coronary heart disease?P ? 0.05?.Conclusion:Through the observation and analysis of prethrombotic state group and non thrombus group of coronary heart disease,we found that TC,TG and hs-CRP may be three clinical parameters that have predictive value for prethrombotic state of coronary heart disease,TC and hs-CRP are significantly correlated with prethrombotic state.The platelet aggregation rate,which is the main predictor of prethrombotic state,has a significant correlation with hs-CRP,suggesting that inflammatory reaction and platelet activation play a key role in the prethrombotic state of coronary heart disease.Part 2 To study the effect and role of Zedoarondiol on RAW264.7 macrophage inflammation model by regulating miR-34a.Objective:To study the effects of Zedoarondiol extracted from Rhizoma Curcumae on inflammatory response and the regulation of miR-34a on the prethrombotic state of coronary heart diseaseMethods:The LPS stimulation model of RAW264.7 cells in vitro was established,the target miR-34a transfection experiment and Zedoarondiol intervention experiment were carried out,and the double luciferase reporter gene analysis was constructed to verify whether Sirtl was the target gene of miR-34a.The experiment was divided into the normal control group?control?,the model group?LPS?,the drug?Zedoarondiol?group?Zed?,the miR-34a group?miR-34a?.the miR-34a +Zedoarondiol group?miR-34a+Zed?,the miR-34a inhibitor group?miR-34a?.34a inhibitor NC group?miR-34a inhibitor NC?,a total of 9 groups,each group of 5 samples.In group Control,only the medium was added.In group LPS,LPS was added to 24h in the medium.In group Zed,Zedoarondiol was added to the medium and 1H was intervened in advance,then LPS was added to interfere with 24h.Group miR-34a was transfected with miR-34a mimic in advance,then LPS was added to interfere with 24h.Group miR-34a+Zed was transferred to miR-34a mimic in advance,then Zedoarondiol was added to 1H in advance,and LPS was added to interfere with 24h.MiR-34a inhibitor group was transferred to miR-34a inhibitor in advance,then LPS was added to 24h.The miR-34a NC group was transfected with miR-34a mimic NC in advance,then LPS was added to interfere with 24h.MiR-34a NC+Zed group was transferred to miR-34a mimic NC in advance,then Zedoarondiol was added to intervene 1h in advance,then LPS was added to 24h.MiR-34a inhibitor NC was transfected into miR-34a inhibitor NC in advance,then LPS was added to interfere with 24h.After the intervention,the expression of inflammatory factors IL-6,TNF-alpha and IL-1 beta in the supernatant of each group was detected by ELISA method.The expression of miR-34a and SirtlmRNA in each cell was detected by RT-PCR,and the protein expression of Sirtl and NF kappa B/TLR-4 was detected by Western-blotting.Results:Compared with the control group,miR-34a showed high expression?P<0.05?in the inflammatory model of LPS stimulated RAW264.7 macrophages.The early intervention of Zedoarondiol could reduce the expression level of the inflammatory factors IL-6,TNF-A and IL-1 beta in the inflammatory model?P<0.05?.It can reduce the level of miR-34a in the model group?P?0.01?,and increase the expression of SirtlmRNA and protein?P?0.01?in the model and reduce the expression of NF kappa B and TLR4 in the downstream inflammatory proteins?P?0.01?.Compared with the control group,the transfected miR-34a group significantly increased the expression level of inflammatory factors IL-6,TNF-alpha and IL-1 beta?P?0.01?,decreased the SirtlmRNA and protein level?P?0.01?,and increased the expression of NF kappa B and TLR4 in the downstream inflammatory proteins?P?0.01,P?0.05?.Compared with the model group,the transfected miR-34a inhibitor group lowered the expression level of miR-34a?P?0.01?,significantly reduced the expression level of inflammatory factors IL-6,TNF-a,IL-1 beta?P?0.01?,and reduced the NF kappa B and TLR4 expression of the downstream inflammatory protein?P?0.01?.In the transfection experiments,the expression of miR34a in the samples of transfected miR-34a mimic was significantly higher than that of the miR-34a mimic NC group and control group?P?0.01?,suggesting that the transfection was successful.Dual luciferase reporter gene analysis showed that miR-34a could act on the 3 'UTR region of Sirtl,and confirmed that Sirtl was the target gene of miR-34a.Conclusion:?1?Zedoarondiol can inhibit the inflammatory response of LPS to RAW264.7 cells;?2?miRNA-34a is highly expressed in the inflammatory model of LPS stimulated RAW264.7,and the inbition of miRN-A-34a expression can inhibit the inflammatory response;?3?Zedoarondiol may inhibit the inflammation by regulating the expression of Sirtl of the target gene of miRNA-34a and the downstream inflammatory pathway. |
PDF Full Text Request