SPLA 2-â…¡ Alentiviruses Express In Human Monocytic Cells And Cause Atherosclerosis

Background:Atherosclerosis disease is one of the principal diseases that threaten human health. At present, the pathogenesy of atherosclerosis disease is not fully studied. Current studies have shown that inflammation plays an essential role in the development and procedure of atherosclerosis disease.Secretory phospholipase A2 of group IIA takes part in the process of atherosclerosis. Inflammation is the main characteristic of atherosclerosis,which participates in the production,progression and the rupture of atherosclerotic plaques. As one of the inflammation factors, secretory phospholipase A2 of group IIA, a subtype of lipoprotein-related phosphatidolipase A2, is mainly produced by smooth muscle cells and macrophages and belongs to calcium-depended phosphatidolipase family, which locates mainly in macrophage gathering place, lipid core area and extracellular matrix of pathological intima. Currently, it's thought to be related with the development of atherosclerosis by possibly hydrolyzing cellular membrane and phosphatidylcholine in lipoprotein to produce proinflammatory reaction and atherosclerotic lysophosphatidylcholine and oxidized fatty acid .Objective:Human sPLA2-â…¡A was synthesized by using the gene sythesis technology in vitro. To construct lentiviruses expressing sPLA2-â…¡A gene. The reconstructed lentiviruses were transfected into human monocytic cells, and research the expression level of sPLA2-â…¡A in human monocytic cells and its function of proatherosclerosis. Methods:Human sPLA2-â…¡A was synthesized by whole-genome synthesis technology according to the gene sequences from gene bank. Then constructed the lentiviruses expressing sPLA2-â…¡A gene. Cultured the human monocytic cells in vitro. The reconstructed lentiviruses were transfected into human monocytic cells. Observed the expression of green fluorescent protein by inverted microscope. The human monocytic cells were collected in the control groups and the experimental groups after 72 hours. The mRNA was extracted, and then reversed transcription into cDNA. Detect the mRNA level by Real-time PCR and the fusion protein by Western blot . All results were statistically analyzed with SPSS 13.0. P<0.05 was considered statistic difference, P<0.01 was considered significantly statistic difference.Results:1. The human sPLA2-â…¡A was synthesized, and the lentiviruses contain sPLA2-â…¡A gene were acquired. The reconstructed lentiviruses of sPLA2-â…¡A were confirmed by the sequence analysis and electrophoresis.2. Cultured the human monocytic cells in vitro, and the cells growed in good condition. The reconstructed lentiviruses of sPLA2-â…¡A were transfected into human monocytic cells. The green fluorescent protein was observed by inverted microscope which indicated the success of lentiviruses transfection.3. The mRNA levels of sPLA2-â…¡A , MCP-1 and ICAM-1 in transfected group increased compared with the control group(P <0.05) .4. The human monocytic cells,which transfected the reconstructed lentiviruses, cultured with ox-LDL,and cultured with ox-LDL and rosuvastatin. The mRNA levels of sPLA2-â…¡A , MCP-1 and ICAM-1in rosuvastatin group decreased compared with the only ox-LDL group(P <0.05) .Conclusions:The sPLA2-â…¡A lentiviruses were constructed successfully and they can be transfected into human monocytic cells. The mRNA levels of sPLA2-â…¡A and inflammatory cytokines in transfected group increased. The human monocytic cells,which transfected the reconstructed lentiviruses, cultured with ox-LDL,and cultured with ox-LDL and rosuvastatin. The mRNA levels of sPLA2-â…¡A and inflammatory cytokines in rosuvastatin group decreased compared with the only ox-LDL group.It demonstrated that ox-LDL can promote arteriosclerosis and rosuvastatin's anti-inflammatory effects.