| BackgroundAtherosclerosis(AS)is the common pathophysiology of ASCVD.Epidemiological studies have shown that AS is highly heritable in many ethnic groups due to many mutations in genes that affect blood lipid levels.The function of these genes can not only affect the level of blood lipids,foam cells and even the inflammatory response,so we can think that the treatment of these genes is an effective way to prevent and treat AS.Proline/serine-rich coiled-coil protein 1(PSRC1),also known as DDA3,is encoded by the PSRC1 gene and is essential for normal mitotic progression of eukaryotic cells,plays an important role for intermediate plate aggregation and mitotic late chromosome normal separation.GWAS study found that PSRC1 gene polymorphism have been associated with serum TC,HDL-C,LDL-C and TG levels in the many population.Thus,PSRC1 and lipid metabolism disorders,and can affect blood fat,affecting the occurrence and development of AS.Our previous study found that the use of serum amyloid substance P(SAP)stimulated mouse macrophages,the cholesterol transfer rate was significantly increased,decreased intracellular cholesterol,cell foam decreased.However,compared with the control group,the PSRC1 gene expression in the SAP-stimulated group was higher than that in the control group.By silencing the PSRC1 gene,the biological effect of anti-macrophage foam of SAP can be reversed.To sum up,domestic and foreign and our previous studies have shown that PSRC1 expression may be related to the occurrence and development of AS.In order to elucidate this hypothesis,this project intends to transfect RAW264.7 cells by adenovirus in vitro and in vivo to observe the effect and mechanism of PSRC1 overexpression on the formation of foam cells;and through the tail vein injection of adenovirus,ApoE-/-mouse PSRC1 gene overexpression,the impact on the occurrence and development of AS and its mechanism.Methods and Results一、In vitroThe adenoviruses(Ad-PSRC1)and control adenovirus(Ad-GFP)overexpressing PSRC1 were constructed and transfected into RAW264.7 cells.The cells were stained with oil red O,HPLC,Elisa,RT-qPCR and Western Bolting and other experimental methods to clarify the PSRC1 overexpression on RAW264.7 cells foam,intracellular cholesterol,the rate of transfer of cholesterol and cholesterol transport protein expression.The study found that PSRC1 overexpression can increase the expression of PPAR-y and LXR-a in RAW264.7 cells on the one hand,increase the cholesterol transfer rate and decrease the content of intracellular cholesterol;on the other hand,IL-1β and IL-6 secretion,thereby inhibiting cell foam.二.In vivoAn ApoE-/-mouse model of PSRC1 overexpression was constructed by tail vein injection of Ad-PSRC1 and Ad-GFP using Oil Red O staining,immunohistochemistry,enzymatic,Elias,RT-qPCR and Western Bolting and other experimental methods to clarify the PSRC1 overexpression on the development of mouse AS and the impact of the body’s macrophages and hepatocytes cholesterol metabolism.The study found that PSRC1 overexpression inhibited the development of AS in ApoE-/-mice with a high fat diet and increased the stability of AS plaque.At the same time,on the one hand,ApoE-/-mice with PSRC1 overexpressing a high-fat diet had lower serum TC/LDL-C/TG levels,higher HDL-C levels and improved HDL function;on the other hand,The levels of serum TNF-α,IL-1β and IL-6 also decreased.Similar to the in vitro studies,overexpression of PSRC1 in liver and peritoneal macrophages up-regulated the expression of PPAR-β and LXR-α.Finally,our study also found that the expression of β-catenin in liver and peritoneal macrophages increased,and the NF-κB pathway was inhibited.ConclusionThis study confirmed that overexpression of PSRC1 in liver and macrophages can play a role in the regulation of cholesterol metabolism and inflammation by increasing the expression of β-catenin,inhibiting the NF-κB pathway,up-regulating the expression of PPAR-y and LXR-a Inhibit the development of AS and increase the role of plaque stability. |
