LOX-1 Regulation in Anti-atherosclerosis of Active Compounds of Herbal Medicine: Current Knowledge and the New Insight / 中国结合医学杂志

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) have recently been identified to be closely related to the occurrence and development of atherosclerosis (AS). A growing body of evidence has suggested Chinese medicine takes unique advantages in preventing and treating AS. In this review, the related research progress of AS and LOX-1 has been summarized. And the anti-AS effects of 10 active components of herbal medicine through LOX-1 regulation have been further reviewed. As a potential biomarker and target for intervention in AS, LOX-1 targeted therapy might provide a promising and novel approach to atherosclerotic prevention and treatment.

Oxidized low-density lipoprotein stimulates dendritic cells maturation via LOX-1-mediated MAPK/NF-κB pathway

Dendritic cells (DCs) play a crucial role as central orchestrators of immune system response in atherosclerosis initiation and progression. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is involved in the immune maturation of DCs, but the underlying mechanisms remain unclear. We isolated mouse bone marrow progenitors and stimulated them with granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4 to induce immature DCs. We then treated DCs with oxidized low-density lipoprotein (oxLDL) to induce maturation. LOX-1 siRNA was used to investigate the modulation of LOX-1 on the development of DCs and the underlying signal pathways. CD11c-positive DCs were successfully derived from mouse bone marrow progenitors. OxLDL promoted the expressions of DCs maturation markers and pro-inflammatory cytokines. OxLDL also upregulated LOX-1 expression and activated MAPK/NF-κB pathways. LOX-1 siRNA could attenuate the expression of MAPK/NF-κB pathways and inflammatory cytokines. In conclusion, oxLDL induced the maturation of DCs via LOX-1-mediated MAPK/NF-κB pathway, which contributed to the initiation and progression of atherosclerosis.

Changes and clinical significance of serum sLOX-1 and omentin-1 in patients with H-type hypertension complicated with acute ischemic stroke / 实用医学杂志

Objective To investigate the changes of soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) and omentin-1 in patients with H-type hypertension complicated with acute ischemic stroke, and to analyze the correlation of sLOX-1 and omentin-1 levels with the severity and prognosis of the disease.Methods Totally 136 patients with H-type hypertension complicated with acute ischemic stroke from February2017 to May 2018 were selected as observation group, and 136 non-acute ischemic stroke patients with H-type hypertension in the same period as the control group. The patients of observation group were divided into mild, moderate and severe sub-groups according to NIHSS score, and they were also divided into good prognosis group and poor prognosis group based on modified RANKIN scale (mRS) score. The serum sLOX-1 and omentin-1 levels were detected, and the correlation of sLOX-1 and omentin-1 levels with severity and prognosis of disease was analyzed. Results The serum sLOX-1 level of the observation group was higher, but the serum omentin-1 level lower than that of control group (P < 0.05). With the severity of the disease, the serum sLOX-1 level increased, but the serum omentin-1 level decreased (P < 0.05). The serum sLOX-1 level of good prognosis group was significantly lower, whereas the serum omentin-1 level significantly higher than that of poor prognosis group (P < 0.05). sLOX-1 was positively correlated with NIHSS score and mRS score, while omentin-1 was negatively correlated with NIHSS score and mRS score (P < 0.05). Conclusions The levels of serum sLOX-1 and omentin-1 are closely related to the severity and prognosis of patients with H-type hypertension complicated with acute ischemic stroke, which could be used as markers for evaluating the severity and prognosis of the patients.

Research of the relevance between serum sLOX-1 levels and left ventricular diastolic function in patients with essential hypertension / 中国综合临床

Objective To investigate the correlation between serum lectin like oxidized low density lipoprotein receptor-1 ( sLOX-1 ) and left ventricular diastolic function in patients with essential hypertension. Methods From January 2016 to July 2017, one hundred and forty-six patients with essential hypertension were selected and divided into two groups according to the ratio of E/ A,the left ventricular diastolic function group (76 cases) and the left ventricular diastolic function group (70 cases),the sLOX-1 level of the patients was detected by ELISA method, and the left ventricular diastolic function was evaluated by echocardiography. The correlation between sLOX-1 and left ventricular diastolic function was analyzed. Results The serum sLOX-1 levels in left ventricular diastolic function group (( 208. 12 ± 13. 48 ) μg/ L ) were significantly higher than those inthe left ventricular diastolic function group ((152. 12 ± 12. 96) μg/ L) . The difference between the two groups was statistically significant (t= 6. 586,P= 0. 000). Logistic regression analysis showed that serum sLOX-1 was a influence factor of left ventricular diastolic function (OR= 2. 42,95%CI 1. 42-2. 82,P = 0. 036) . Conclusion The levels of serum sLOX-1 can be used as a indicator of left ventricular diastolic function.

Study on correlation between serum sFas and sLOX-1 with occurrence and development of acute coronary syndrome / 重庆医学

Objective To investigate the correlation between serum sFas and sLOX-1 with occurrence and development of acute coronary syndrome (ACS).Methods A total of 52 patients definitely diagnosed ACS (ACS group) by coronary artery angiography (CAG) were enrolled,including 23 cases of unstable angina (UA group) and 29 cases of acute myocardial infarction (AMI group),and contemporaneous 58 cases of non-coronary arterial stenosis confirmed by CAG were selected as the control group (NC group).The serum levels of sFas and sLOX-1 were measured by enzyme linked immunosorbent assay.Results Compared with the NC group,the serum levels of sFas and sLOX-1 in the ACS group were increased,the serum levels of sFas and sLOX-1 in the AMI group and UA group were higher than those in the NC group,moreover which in the AMI group were higher than those in the UA group (P＜0.01).The serum sFas level in the ACS group was positively correlated with the sLOX-1 level (r=0.825,P=0.001),but both had no obvious correlation with the serum levels of CK-MB and cTnⅠ (P＞0.05).Conclusion High levels of serum sFas and sLOX-1 may be the risk factors of ACS.

Prevention of 2 ,3 ,5 ,4 '-Tetrahydroxystilbene-2-O-beta-D-Glucoside on Atherosclerotic Plaque Formation in ApoE Knock-out Mice / 医药导报

Objective To observe the protective effect and mechanism of 2,3,5,4 '-tetrahydroxystilbene-2-O-beta-D-glucoside ( THSG) on atherosclerosis in ApoE konck-out mice. Methods A total of 24 ApoE knock-out mice were randomly divided into normal control group (n=8), model control group (HFD, high-fat diet, n=8) and treated group (THSG, 20 mg· kg-1, i. g. , n=8). The atherosclerosic plaque of aorta wall and aorta root were measured by oil red O staining;The expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in human umbilical vein endothelial cells (HUVEC) through C-reaction protein ( CRP) was studied by Western blotting. Results The atherosclerosis plaque in normal control group was not observed. The lipid accumulation decreased in the aorta and the plaque areas in the aortic sinus in THSG treated-group compared with model control group. Moreover, THSG down-regulated CRP-induced LOX-1 expression in HUVEC. Conclusion The atheroscletosis plaque in ApoE knock-out mice was decreased by THSG. The mechanism might be related to the inhibition of the expression of LOX-1 protein.

Study on effects of Shenqi Mixture on blood vessel protection in patients with different types of hyperlipidemia / 重庆医学

Objective To observe the effects of Shenqi Mixture on sLOX‐1 ,SOD ,MDA ,NO ,ET and lipid level among the patients with different types of hyperlipidemia and to investigate the mechanism of blood vessel protection .Methods Sixty‐four patients with hyperlipidemia were divided into high TC group (20 cases) ,high TG group(12 cases) ,mixed type hyperlipidemia group(23 cases) and low HDL group(9 cases) .Every patients were treated with Shenqi Mixture 10 g/d ,treatment course was 6 weeks .Other 25 healthy people of normal blood lipid were selected as the control group .The levels of sLOX‐1 ,SOD ,MDA ,NO ,ET and blood lipid were detected before and after treatment .Results The levels of sLOX‐1 was increased by high TC and mixed type of hyperlipidemia(P0 .05) .Conclusion Shenqi Mixture has good effects on blood vessel protection and protects the vascular endothelial cells ,thus plays the protective role on blood vessel .

Study of Xuefu Zhuyu Decoction anti-atherosclerotic mechanism / 中国免疫学杂志

Objective:To investigate the influence of Xuefu Zhuyu Decoction ( XZD) contained Drug-Serum on the expression of TLR4/NF-κB signals and LOX-1, TNF-α, ICAM-1, VCAM-1 in HUVECs, and to study its possible anti-atherosclerotic mechanism.Methods:HUVECs was cultured in vitro and divided equally into the normal group ,the model group,the ATV group and the XZD group in random.HUVECs were stimulated with LPS for 2 h,then treated separately with the drug-serum and atorvastatin for 24 h,finally measured the expression of TLR 4, MyD88, TRAF-6, NF-κB, LOX-1, TNF-α, ICAM-1 and VCAM-1 mRNA with real-time PCR,the expression of TLR4,NF-κB,LOX-1,TNF-α,ICAM-1 and VCAM-1 protein were analyzed by Western blot method .Results:Protein and mRNA expressions of TLR4,MyD88,TRAF-6,NF-κB,LOX-1,TNF-α,ICAM-1 and VCAM-1 increased significantly after LPS stimulation(vs normal control group,P<0.01),Xuefu Zhuyu Decoction decreased the high expression of TLR 4,MyD88,TRAF-6, NF-κB,LOX-1,TNF-α,ICAM-1 and VCAM-1(vs model group,P<0.05 ).Conclusion:Xuefu Zhuyu Decoction can block the high ex-pression of TLR4 and its downstream signal transduction pathway and the high expression of LOX -1, TNF-α, ICAM-1 and VCAM-1.Maybe it′s the mechanism of Xuefu Zhuyu Decoction exert the function of anti-artherosclerosis.

Immune maturation of human monocyte-derived dendritic cells induced by angiotensinⅡ / 中国病理生理杂志

AIM:To investigate the effects of angiotensin II ( Ang II) on the immune maturation and the oxi-dized low-density lipoprotein (Ox-LDL)-uptaking capacity of human monocyte-derived dendritic cells (DCs).METH-ODS:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation , and the monocytes were purified by positive selection with anti-CD14 magnetic beads.After cultured with rhGM-CSF (100 μg/L) and rhIL-4 (50μg/L) for 5 d, the monocytes differentiated into immature DCs .On the 6th day of the culture, the cells were treated with various concentration levels of Ang II or pretreated with losartan .The immunophenotypic expression of HLA-DR and CD83 was analyzed by flow cytometry .The secretion levels of IL-12 and IFN-γin the culture supernatants were measured by ELISA.Furthermore, DCs were incubated with DiI-labelled Ox-LDL.The DiI-Ox-LDL-incorporated fraction was investiga-ted by flow cytometry .The mRNA expression of 3 scavenger receptors , scavenger receptor A ( SR-A) , CD36 and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), was examined by real-time PCR.RESULTS: Ang II induced the maturation of human monocyte-derived DCs, stimulated the expression of CD83 and HLA-DR, and promoted the secre-tion of IL-12 and IFN-γ, which were suppressed by losartan .Furthermore, Ang II increased the Ox-LDL-uptaking capacity of DCs, which was partially reduced by losartan .The incubation of DCs with Ang II enhanced the mRNA expression of LOX-1 in a dose-dependent manner , which was reduced by losartan .However, the expression of SR-A and CD36 was not changed .CONCLUSION:Ang II promotes the immune maturation of human monocyte-derived DCs and increases the up-take of Ox-LDL probably through the up-regulation of LOX-1 expression.

The significance of soluble lectin-like oxidized low-density lipoprotein receptor-1 on early diagnosis of acute ST elevation myocardial infarction / 中国综合临床

Objective To investigate whether soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) as the early diagnostic biomarker of acute ST elevation myocardial infarction (STEAMI).Methods Sixty-five patients with STEAMI and 30 patients with stable coranary heart disease or other heart disease(control group) were enrolled as our subjects.Serum sLOX-1 levels were measured.Results The median(P25,P75) of Serum sLOX-1 in the patients with STEMI were 210.0 (130.0,356.0) ng/L,significantly higher than that of control group(65.5 (55.2,85.2) ng/L,Z =6.17,P ＜ 0.001).Logistic regression analysis revealed that sLOX-1 alone was an independent factor associated with STEAMI (B =0.036,P ＜ 0.001).The area under the ROC curve of sLOX-1 for detecting STEMI was 0.895,and 95％ CI was 0.831-0.959 (P＜0.001).Taking sLOX-1 =87.5 ng/L as cut-off value,the sensitivity was 89.6％ and specificity was 82.4％for the diagnosis of STEAMI.Conclusion Serum sLOX-1 was significantly higher in the STEAMI and it might served as the early diagnostic marker for STEAMI.

Protective Effect of Angiotensin Converting Enzyme 2 Transfection on Vascular Endothelial Cells / 中国循环杂志

Objective: To investigate the inlfuence of angiotensin converting enzyme 2 (ACE2) on lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) protein expression and to explore the protective effect of ACE2 on vascular endothelial cells. Methods: Our work includedin vitro andin vivo studies. For in vitro experiment, the human umbilical vein endothelial cell (HUVEC) were cultured and transfected with replication deficient recombinant adenovirus of ACE2 (Ad-ACE2), and LOX-1 protein expression stimulated by angiotensin 2 was examined by Western blot analysis. Forin vivo study, atherosclerosis plaques were induced in 20 apolipoprotein E-deifcient (ApoE-/-) mice, and then randomly divided them into 2 groups: ACE2 group, the mice received Ad-ACE2 (2.5×109 pfu/ml) injection through caudal vein, EGFP (enhanced green lfuorescent protein) group, the mice received equal replication deifcient recombinant adenovirus of EGFP (Ad-EGFP) injection through caudal vein.n=10 in each group. The animals were executed after 1 month treatment to collect abdominal aorta. Lipid content in atherosclerosis plaque was evaluated by Oil red O staining and LOX-1 protein expression was examined by immunohistochemistry and Western blot analysis. Results: Bothin vitro andin vivo experiments conifrmed that endothelial cell LOX-1 protein expression was signiifcantly inhibited by ACE2 transfection. The lipid content in ACE2 group was obviously lower than that in EGFP group byin vivo study. Conclusion: ACE2 may inhibit LOX-1 protein expression and therefore reduce the progress of atherosclerosis.

Effects of rosuvastatin on atherosclerosis and LOX-1, NF-κb p65 expression in the aorta of apoe-deficient mice / 解放军医学杂志

Objective To investigate the effects of HMG-CoA reductase inhibitor rosuvastatin on atherosclerosis and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), and nuclear factor (NF)-kB p65 expressions in apolipoprotein E (ApoE)-deficient mice. Methods Twenty six-week-old ApoE-deficient male mice were randomly divided into hyperlipidemia model group (n=10) and rosuvastatin group (n=10), and they were fed high-fat diet for 13 weeks. Ten six-week-old C57BL/6J (wild type, WT) male mice were selected as normal control group, and were fed normal diet for 13 weeks. After 13 weeks, blood was drawn from the mice to determine serum levels of total cholesterol (TCH), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C). These mice were sacrificed, and their aortas were obtained and examined with HE staining; Western blotting and RT-PCR were used to analyze LOX-1, NF-kB p65 expression intensity in aorta tissue quantitatively. Results The serum level of TCH, TG and LDL-C in rosuvastatin group were lower than those in hyperlipidemia model group (P<0.05). Pathological observation showed that atherosclerotic lesions of the aortas were aggravated significantly in hyperlipidemia model group but alleviated in rosuvastatin group compared with normal control group. Compared with normal control group, LOX-1, NF-kB p65 protein and mRNA expressions significantly increased in hyperlipidemia model group (P<0.05) and reduced in rosuvastatin group (P<0.05). Conclusions Rosuvastatin may lower blood lipid significantly, alleviate the degree of atherosclerotic lesions, and inhibit LOX-1, NF-kB p65 protein and mRNA expressions in the aortic tissue of ApoE-deficient mice. Its anti-athrosclerosis effect is related to down regulation of LOX-1 and NF-kB p65 expressions.

Lox-1, a new target in cardiovascular disease / 药学实践杂志

Objective Lox-1 ( lectin-like oxidized low-density lipoprotein receptor-1 ) was one of the main receptor of ox-LDL, which took an important role in vascular endothelial dysfunction , the formation of foam cells , and the stability of atherosclerotic plaques .To investigate the role of Lox-1 in cardiovascular diseases .Methods The literatures of Lox-1 in cardiovascular diseases retro-spectively were reviewed .Results As a new ox-LDL scavenger receptor , Lox-1 played an important role in cardiovascular diseases . Conclusion Lox-1 might provide new ideas for cardiovascular diseases .

Different Effects of Simvastatin on the Expression of Lectin-like Oxidized Low-Density Lipoprotein Receptor-1 (LOX-1) Induced by Oxidized Low-Density Lipoprotein (ox-LDL) or Glucose / 天津医药

Objective To investigate the effects of simvastatin on the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) induced by oxidized low-density lipoprotein (ox-LDL) or Glucose in U937 macrophages, and explore the role of NF-κB in modulating of LOX-1 expression. Methods U937 macrophages were treated with PMA to induce differentiation, which were co-cultured with 50 mg/L ox-LDL or/and 25 mmol/L glucose. Pyrrolidine dithiocarbamate (PDTC) and simvastatin (1 μmol/L or 10 μmol/L) were used to treat cells. The expression of LOX-1 protein and NF-κB ac- tivity were detected by enzyme-linked immunosorbent assay technology. The expression of LOX-1 mRNA was measured by RT-PCR. Results The expression of LOX-1 was up regulated by ox-LDL, glucose and combination of both. The inhibitor of NF-κB PDTC suppressed this up-regulation. Simvastatin suppressed the expression of LOX-1 induced by ox-LDL, and showed a significant effect in the higher concentration. There was no significant effect of simvastatin on the expression of LOX-1 induced by glucose. The variation of NF-κB activity was similar to that of LOX-1 expression. Conclusion Simvas- tatin suppressed the expression of LOX-1 induced by ox-LDL, while no significant effect on the expression of LOX-1 in- duced by glucose. The expression and regulation of LOX-1 were related with NF-κB pathway.

Role of oxLDL/LOX-1 system and NF-κB pathway in early diabetic vascular endothelial dysfunction / 中华内分泌代谢杂志

Diabetic rat model was established by peritoneal injection of streptozocin.At the end of 2 weeks,oxidized low-density lipoprotein (oxLDL) level in diabetic rats was raised [ ( 2.87 ± 0.40 vs 2.27 ± 0.36 ) μg/dl,P＜0.05 ] and endothelium-dependent relaxation was sluggish compared with normal rats.At the end of 6 weeks,oxLDL level continued to increase [ 4.32 ±0.66 ) μg/dl,P＜0.01] and endothelium-dependent maximum relaxation ( Rmax ) was decreased obviously ( P ＜0.01 ).Meanwhile,the protein and mRNA expressions of lectin-like oxidized lowdensity lipoprotein receptor-1 ( LOX-1 ),NF-kB,and ICAM-1 on vessel wall of diabetic rats were higher than those in normal rats,and LOX-1 mRNA was positively correlated with the levels of oxLDL,NF-kB,and ICAM-1 mRNA,while negatively correlated with Rmax,indicating that OxLDL/LOX-1 system may cause early endothelial dysfunction in diabetes via activating NF-kB and up-regulating ICAM-1 expression.

Modulation of cardiac fibroblast function by thiazolidinediones with a focus on the role of LOX-1 / 中南大学学报(医学版)

Cardiac fibroblasts play a pivotal role in cardiac remodeling.In response to various pro-fibrotic stimuli,such as pro-inflammatory cytokines,anoxia-reoxygenation and pressure overload as well as aging,cardiac fibroblasts undergo proliferation,migration and activation,leading to the accumulation of extracellular matrix components and increased thickness and stiffness of heart.The ligands for peroxisome proliferator-activated receptor γ,especially thiazolidinediones,modulate the function of cardiac fibroblasts and the progression of cardiac remodeling,especially under pathological conditions.Unfortunately these agents have not been found to be consistently beneficial in heart failure.Although the precise intracellular signaling pathways are not fully understood,existing evidence strongly supports the involvement of oxidative stress and related signaling pathways.Further,peroxisome proliferator-activated receptor γ and lectin-like oxidized low-density lipoprotein receptor-1 together play critical roles in thiazolidinediones-modulated cardiac fibroblast dysfunction.

Oxidized low density lipoprotein-induced expression of transforming growth factor beta 1 and production of extracellular matrix in rat glomerular mesangial cells is mediated by lectin-like oxidized low density lipoprotein receptor-1 / 第二军医大学学报

Objective: To investigate the effect of oxidized low-density lipoprotein (ox-LDL) on expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), transforming growth factor beta 1 (TGF-β 1), and secretion of extracelluar matrix (ECM) in cultured rat glomerular mesangial cells (GMCs), and to investigate the influence of LOX-1 inhibitor polyinosinic acid (PIA) on the effect of ox-LDL. Methods: Rat glomerular masengial cells were cultured in vitro. RT-PCR was employed to determine the LOX-1 mRNA expression in GMCs incubated with different concentrations of ox-LDL (0, 25, 50, 100 μg/ ml). The expression of LOX-1 and TGF-β1, mRNA was also determined by RT-PCR in the blank control group, ox-LDL (50 μg/ml) group and PIA(50 μg/ml ox-LDL+250 μg/ml PIA) group. The contents of TGF-β1 fibronectin (FN), and collagen IV (Col IV) in the supernatants of the above 3 groups were determined by ELISA. Results: RT-PCR showed that LOX-1 mRNA expression in 25, 50 and 100 μg/ml ox - LDL groups was significantly higher than that of blank control group(P<0.05), with the highest expression found in the 50 μg/ml ox-LDL group; the expression of LOX-I and TGF-β 1 mRNA was significantly higher in 50 μg/ml ox-LDL group than that in the other 2 groups (P< 0.01). ELISA results demonstrated that the supernatant contents of TGF-β1, FN and Col IV were significantly higher in 50 μg/ml ox-LDL group than those in the other 2 groups (P< 0.05). Conclusion: Ox-LDL can upregulate the expression of LOX-1 and TGF-β1, mRNA and the secretion of extracelluar matrix in GMCs. Polyinosinic acid can antagonize the above effect of ox-LDL, suggesting that LOX-1 may participate in ox-LDL-induced GMCs damage and is involved in the development and progression of glomerulosclerosis.

Lectin-Like Oxidized Low-Density-Lipoprotein Receptor-1 and Atherosclerosis / 国际脑血管病杂志

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a specific receptor of oxidized low-density lipoprotein (oxLDL) expressed in endothelial cells. It is a type Ⅱ single-chain transmembrane protein and belongs to C-type lectin family. LOX-1 leads to the injury of endothelial cells by mediating oxLDL, and plays a role in the initiation and progression of atherosclerosis, hypertension, thrombosis. This article reviews the association between LOX-1 and atherosclerosis.

Effects of fluvastatin and Shexiang baoxin pill on expression of 1 lectinlike oxidized low density lipoprotein receptor-1 in a rabbit model of atherosclerosis / 重庆医科大学学报

Objective:To explore the expression of LOX-1(lectin like oxidizeol low density lipoprotein receptor-1,LOX-1) in aorta of atherosclerotic rabbits and modulating the effect of fluvastatin and Shexiang baoxin pill on the gene and protein expression of LOX-1. Methods:Tirty-two New Zealand white rabbits were randomly divided into four groups: hypercholesterol diet group (fed mth high cholesterol diet),hypercholesterol diet plus Fhevastatin group(fed with high cholesterol diet plus Fhe vastain 25mgl kgld).hypercholesterol diet plus shexiang Bao xin Pill group (fed with high cholesterol diet plus shegxiang Baoxinpill 25mgl kgld).normal diet gwup(fed mith regnlar chow),the protein expression of LOX-1 in aorta of rabbits were examined by immunohistochemsthy,the gene expresswin of LOX-1 in aorte of rabbits were examined by R7-PCK. Results:hypercholesterol diet group(fed with high cholesterol diet),hypercholesterol diet plus fluvastatin group(fed with high cholesterol diet plus Fluvastatin 25mg/kg?d), hypercholesterol diet plus Shexiang baoxin pill group(fed with high cholesterol diet plus Shexiang Baoxin Pill 25mg/kg?d),normal diet group(fed with regular chow),the protein expression of LOX-1 in aorta of rabbits were examined by immunohistochemistry,the gene expression of LOX-1 in aorta of rabbits were examined by RT-PCR. Conclusion:LOX-1 was expressed in rabbit aorta of hypercholesterol diet group and two treatment groups, expression of LOX-1 in two treatment groups both significantly reduced compared with hypercholesterol diet group(P

A Study in Construction of Short Hairpin Small Interfering RNA Expression Vector Target Lectin Like Oxidized Low Density Lipoprotein Receptor-1 Gene and Its Effect on Foam Cells / 中国生物工程杂志

Aim:To construct the short hairpin small interfering RNA(shRNA) eukaryotic expression vector specific to mouse lectin like oxidized low density lipoprotein receptor 1(LOX-1) gene and to observe its silencing effect on LOX-1 in RAW264.7 cells.Methods:(1)The pLOX-1-shRNA expression vector was constructed by gene recombination,Then transfected into the cultured RAW264.7 cells.At 48 h after Transfection,the expression of LOX-1 mRNA in RAW264.7 cells were determined by semi-quantitative RT-PCR,the expression of LOX-1 proteins examined by Western blot.(2) Oil Red O Dyeing experiment was used to show the cellular lipid droplets in lipid-loaded cells.The method of cholesterol oxidase analysis was performed to determine the content of cellular cholesterol.The ability of uptake Dil-ox-LDL in RAW264.7 cells was assayed by fluorescence microscopy.Results: pLOX-1-shRNA expression vector was successfully constructed.Transfection of pLOX-1-shRNA expression vector into RAW264.7 cells down regulaled the expression level of LOX-1 gene,as compared with the control group,transfection of the RAW264.7 cells with LOX 1-shRNA led to a remarkable reduction of the number macrophages transformed into foam cell,and could suppress the uptake of ox-LDL.Conclusion:The pLOX-1-shRNA expression vector can inhibit the expression of LOX 1 in RAW264.7 cells and the transformation of the macrophages into foam,which may he beneficial in searching new gene therapy of atherosclerosis.