| Objective: To establish an atherosclerotic vulnerable plaque model in New Zealand white rabbits with balloon injury and fed high-fat diet,and analyzed the expression difference lnc RNA TCONS＿02443383 in abdominal aortic plaque tissue of New Zealand rabbits in normal group and model group,to explore the correlation between lnc RNA TCONS＿02443383and atherosclerotic vulnerable plaque.Synthesis of lnc RNA TCONS＿02443383 overexpressed lentivirus,lnc RNA TCONS＿02443383 overexpressed lentivirus was injected through ear marginal vein and increased the level of lnc RNA TCONS＿02443383 in atherosclerotic vulnerable plaque in New Zealand white rabbits,to observe the stability of atherosclerotic plaque is changed by lnc RNA TCONS＿02443383overexpression lentivirus.Transcriptome sequencing was performed on the abdominal aorta of New Zealand white rabbits.The sequencing data were bioinformatics analyzed.The molecular mechanism of lnc RNA TCONS＿02443383 in the occurrence and development of atherosclerosis was analyzed in combination with laboratory examination and pathological comparison.Methods: Through the abdominal aortic puncture approach of New Zealand rabbits,the abdominal aortic intima of New Zealand rabbits was injured by balloon and fed with high-fat diet to construct the vulnerable plaque model of abdominal main atherosclerosis in New Zealand rabbits.The model was divided into low-fat control group(low-fat feeding without any treatment),model control group,lnc RNA TCONS＿02443383 overexpression group,lnc RNA TCONS＿02443383overexpression control group,lnc RNA TCONS＿02443383 overexpression of lentiviru swas synthesized after successful modeling,lnc RNA TCONS＿02443383 overexpression group was injected through ear marginal vein lentivirus was injected once a week for two weeks.The New Zealand white rabbits in each group were killed after high-resolution magnetic resonance examination of vascular wall,and the blood was taken to detect IL-4,IL-6,IL-8 and TNF-α by ELISA;The abdominal aorta of New Zealand white rabbits was stained with HE staining,oil red O staining and Masson staining to observe the pathological changes.The expression level of lnc RNA TCONS＿02443383 in the four groups of rabbits was detected by real-time fluorescence quantitative PCR,the levels of apoptotic proteins Caspase-3,Bax and Bcl-2 were detected by Western blot,and the contents of TC,TG,HDL and LDL in serum were detected by biochemical detection kit.The transcriptome was sequenced and the sequenced data were bioinformatics analyzed.Results:(1)q RT-PCR: the results showed that the expression of lnc RNA TCONS＿02443383 gene decreased significantly in the model control group,and the lnc RNA TCONS＿02443383 overexpression group increased significantly compared with the model control group(P<0.05).(2)ELISA test: the results showed that IL-6,IL-8 and TNF-ɑ were all increased in the model control group(P<0.05),the levels of IL-4,IL-6,IL-8 were all decreased in lnc RNA TCONS＿02443383 overexpression group(P<0.05).The levels of TNF-ɑ was increased in the model control group(P<0.05).(3)Western blot:the results showed that compared with the low-fat control group,the expressions of Caspase-3 and Bax were up-regulated in the model group(P < 0.05),and down-regulated in the lnc RNA TCONS＿02443383 overexpression group(P < 0.01).(4)Biochemical test results showed that compared with the control group,the model control group significantly increased the contents of TG,TC and LDL,and decreased the contents of HDL.Compared with the model control group,the lnc RNA TCONS＿02443383overexpression group decreased TG,TC and LDL,and increased HDL levels.Compared with the lnc RNA TCONS＿02443383 overexpression group,the lnc RNA TCONS＿02443383 overexpression control group increased TG,TC and LDL levels,and decreased HDL levels.(5)Tissue HE staining: the results showed that there was no abnormal change in abdominal aorta of rabbits in low-fat control group;In the model control group and the lnc RNA TCONS＿02443383 overexpression control group,the abdominal aorta endothelium was thickened,and there were lipid deposition and foam cells in the intimal surface,and fibrous tissue proliferation in the vascular wall.In the lnc RNA TCONS＿02443383 overexpression group,the abdominal aorta endothelium was thickened and the fibrous tissue was proliferated,but the hemorrhage and necrosis were significantly improved.(6)The deposition of lipid droplets in abdominal aorta was observed by oil red O staining.Compared with the low-fat control group,the amount of lipid droplets on the surface of abdominal aortic vascular tissue in the model control group was more,and the difference was statistically significant(P<0.001).Compared with the model group,the amount of lipid droplets on the surface of abdominal aortic vascular tissue in the lnc RNA TCONS＿02443383 overexpression group was less,and there was significant difference between the two groups(P<0.001).Compared with lnc RNA TCONS＿02443383 overexpression group,lnc RNA TCONS＿02443383 overexpression control group had more lipid droplets on the surface of abdominal aorta tissue,which was statistically significant(P<0.001).(7)Masson trichrome staining showed that compared with the low-fat control group,there was fibrosis on the surface of abdominal aorta tissue in the model control group,the vascular fibrosis in the lnc RNA TCONS＿02443383overexpression group was less than that in the model group,and the vascular fibrosis in the lnc RNA TCONS＿02443383 overexpression control group was also obvious.Compared with the low-fat control group,the fibrosis area of the model control group and the lnc RNA TCONS＿02443383 overexpression control group increased significantly(P <0.001,P < 0.01).Compared with the model control group,lnc RNA TCONS＿02443383overexpression significantly improved fibrosis(P < 0.001).(8)Transcriptome sequencing bioinformatics analysis of sequencing data identified that lnc RNA TCONS＿02443383was overexpressed in the New Zealand white rabbit model of atherosclerotic vulnerable plaque,which may inhibit the development of atherosclerosis by up regulating the expression of PPAR signal pathway and cell adhesion related genes and up regulating the expression of lnc RNA TCONS＿02443383 related to overexpression of lnc RNA TCONS＿02443383.Conclusion: After constructing the New Zealand white rabbit atherosclerotic vulnerable plaque model,the expression level of lnc RNA TCONS＿02443383 gene in New Zealand white rabbits decreased significantly,and the injection of lnc RNA TCONS＿02443383 overexpression lentivirus through the ear margin vein can enhance the expression level of lnc RNA TCONS＿02443383 in New Zealand white rabbits,improve the atherosclerosis of New Zealand white rabbits,and promote the stability of plaque.The abdominal aorta of New Zealand rabbits was taken for transcriptome sequencing,and the sequencing data were analyzed bioinformatics.It was identified that the overexpression of lnc RNA TCONS＿02443383 in the atherosclerotic New Zealand rabbit model may inhibit the development of atherosclerosis and promote plaque stability by up regulating PPAR signaling pathway and cell adhesion related genes and up regulating the expression of lnc RNA related to overexpression of lnc RNA TCONS＿02443383.. |
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