Study On A Recombinant HDL Anti-atherosclerosis Nanoliposome Assembled By ApoA-? Mimetic Peptide And Tanshinone ?A

FAMP5?Fukuoka University apoA-I mimetic peptide type5?is a mimetic peptide of apolipoprotein apoA-I synthesized by Fukuoka University,Japan.Studies have shown that FAMP5 has an effect on promoting cholesterol efflux and reducing vascular plaque area.Tanshinone IIA?TA?has a certain improvement effect on coronary blood circulation and vascular endothelial cells,so it has a certain therapeutic effect on anti-atherosclerotic effect.However,because of the low solubility in water,short half-life,and low bioavailability of TA,its clinical application is limited.Recombinant HDL can effectively regulate lipid metabolism disorder caused by hyperlipidemia and effectively prevent and improve atherosclerosis.At the same time it can also be used as a drug carrier to encapsulate hydrophobic drugs in the lipid core.In order to solve the indissolvable problems of TA,and the costly and complicated extraction process of apolipoprotein,this study designed and prepared discoidal and spherical biomimetic high density lipoproteins?TA-d-rHDL and TA-s-rHDL?nanoliposomes.In the new nanoliposomes,the apos was replaced with short apoA-I mimetic peptide FAMP5,and the poorly soluble TA was packaged into the lipid core of rHDL.The promoting cholesterol efflux effects,the reducing vascular plaque area effects and the biocompatibility of the new nanoliposomes were evaluated.FAMP5 and FAMP5-ACD were successfully obtained by solid phase synthesis and HPLC purification with a purity of 91.22%and 92.26%,respectively.The novel TA-d-rHDL and TA-s-rHDL were prepared by thin-film dispersion and nanoprecipitation/solvent evaporation.The physical and chemical characterization of rHDL was determined.Transmission electron microscopy?TEM?and scanning electron microscopy?SEM?shows that TA-d-rHDL and TA-s-rHDL appeared discoidal and spherical nanoparticles,respectively.And they are in uniform size and distribution.The mean diameters,zeta potentials and PDI of the TA-s-rHDL were 100nm,-9.38 and 0.304 measured by Malvern particle size analyzer.The encapsulation efficiency of TA in TA-d-rHDL and TA-s-rHDL was 84.46±0.016%and 80.35±0.06%,respectively.The binding efficiency of FAMP5 in TA-d-rHDL and TA-s-rHDL was 60.88±0.92%and 85.08±0.71%,respectively.The stability experiment showed that the appearance of TA-L,TA-NLC,TA-d-rHDL and TA-s-rHDL did not change within 2 months.At the same time,the encapsulation efficiency of TA decreased from84.46±0.016%and 80.35±0.06%to 81.76±0.122%and 77.14±0.099%respectively,did not change substantially,indicating that the nanoliposome were relatively stable.In order to study the anti-atherosclerosis effect of TA-rHDL in rats,forty-nine Wistar rats were used and randomLy divided into 7 groups,which were named normal group,model group,TA+FAMP5 group,TA-NLC group,s-rHDL group,low concentration TA-s-rHDL group and high concentration TA-s-rHDL group.In addition to the normal group,the other groups were given a fixed amount of high-fat emulsion at a fixed time every day,and intraperitoneal injection of VD₃was conducted according to the time.After 28 days,the rats were given the drug accordingly in groups for 30 days.Finally,the blood lipids and biochemical indexes of serum and liver tissues of each group of rats were measured,and pathological sections of liver and aorta were observed.Serum and liver biochemical indicators show that TA-s-rHDL has certain anti-inflammatory and anti-oxidative effects and it has the effects of reducing TC,TG and LDL and improving HDL.Pathological sections showed that TA-s-rHDL has certain anti-atherosclerosis and has the effect of prevention fatty liver.In this study,TA,FAMP5 and rHDL were combined,and apoA-I was replaced by FAMP5 in an innovative way,making rHDL not only serve as a drug carrier,but also play an anti-atherosclerosis effect.RHDL also increases the amount of TA that goes into the body to work.The results showed that the effect of TA-s-rHDL against AS was doubled and the liver-protecting effect was stronger.As a whole,it has played multiple functions to achieve safe,efficient,simple and stable drug delivery.This study is expected to be applied to the modification and delivery of hydrophobic anti-atherosclerosis drugs.This study provides a useful exploration for good anti-AS drug delivery.