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.

Prediction of poor prognosis in patients with acute cerebral infarction / 中国医师进修杂志

Objective:To explore the predictive values of serum monocyte chemoattractant protein-1 (MCP-1), high mobility protein B1 (HMGB1), adiponectin (APN) and oxidized low density lipoprotein (ox-LDL) levels on poor prognosis of patients with acute cerebral infarction(ACI).Methods:One hundred and sixty-fivepatients with ACI in Zibo Hospital, Shandong Guoxin Nursing Group from October 2018 to December 2020 were selected as the observation group, and 147 healthy people in the same period were selected as the normal control group. The levels of serum MCP-1, HMGB1, APN and ox-LDL were detected. In addition, the observation group was followed up for 3 months after discharge, and the observation group was divided into good prognosis group and poor prognosis group by modified Rankin Scale score. The levels of serum MCP-1, HMGB1, APN and ox-LDL between the poor prognosis group and the good prognosis group were compared. The influencing factors of poor prognosis in patients with ACI and the predictive value of serum MCP-1, HMGB1, APN and ox-LDL levels on poor prognosis were analyzed by Logistic multiple regression analysis and receiver operating characteristic (ROC) curve.Results:The levels of serum MCP-1, HMGB1 and ox-LDL in the observation group were higher than those in the normal control group: (322.61 ± 65.27) ng/L vs. (163.18 ± 15.12) ng/L, (6.61 ± 3.54) μg/L vs. (2.90 ± 0.41) μg/L, (481.11 ± 177.67) mg/L vs. (247.47 ± 27.13) mg/L; but the level of serum APN was lower than that in the normal control group: (10.63 ± 3.80) μg/L vs. (17.65 ± 2.87) μg/L, there were statistical differences ( P<0.05). After 3 months of follow-up, the incidence rate of poor prognosis in the observation group was 35.15%(58/165). The serum levels of MCP-1, HMGB1 and ox-LDL in the poor prognosis group were higher than those in the good prognosis group: (372.15 ± 71.33) ng/L vs. (295.76 ± 42.23) ng/L, (9.74 ± 3.96) μg/L vs. (4.91 ± 1.62) μg/L, (631.03 ± 196.84) mg/L vs. (399.85 ± 95.07) mg/L; but the serum APN level was lower than that in the good prognosis group: (7.62 ± 2.83) μg/L vs. (12.27 ± 3.22) μg/L, there were statistical differences ( P<0.05). The results of Logistic multiple regression analysis showed that age, hypertension, hyperlipidemia, infarct volume, nerve function defect score, time from onset to treatment and MCP-1, HMGB1, APN and ox-LDL levels were the influencing factors of poor prognosis in patients with ACI ( P<0.05). The results of ROC curve analysis showed that the sensitivity and area under the curve of serum MCP-1, HMGB1, APN and ox-LDL levels in combined predicting the poor prognosis were 98.28% and 0.954, which were higher than the single index evaluation ( P<0.05). Conclusions:The serum levels of MCP-1, HMGB1 and ox-LDL are closely related to the prognosis of ACI patients, and all of them have a certain predictive value for the poor prognosis of patients, but the combined prediction efficiency of four items is more higher.

Guanxinning Tablet and their active substances alleviate ox-LDL-induced endothelial cell injury / 药学学报

This study investigated the intervention effect of Guanxinning Tablet on human umbilical vein endothelial cells (HUVECs) injury induced by oxidized low density lipoprotein (ox-LDL), providing experimental basis for Guanxinning Tablet in the treatment of atherosclerosis-related diseases. Under the damage of HUVECs by ox-LDL, the cell viability was detected by CCK-8 (cell counting kit-8) assay; lactate dehydrogenase (LDH) in the cell culture supernatant was detected by the corresponding kit; the cell morphology of different groups was observed by common phase contrast microscope; reactive oxygen species (ROS) and NO levels in the cells were detected by DCFH-DA and DAF-FM DA probes, respectively; monocyte adhesion assay was used to detect the recruitment of THP-1 in HUVECs, and TMRM dye was used to detect the level of mitochondrial membrane potential; interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) secretion in the cells was detected by ELISA assay. The results showed that Guanxinning Tablet had a concentration-dependent proliferative effect on HUVECs. Under the stimulation of 100 μg·mL-1 ox-LDL, the morphology of endothelial cells was significantly changed. At this time, NO level was significantly decreased, ROS level was significantly increased and accompanied by a decrease in mitochondrial membrane potential. The recruitment of THP-1 cells by endothelial cells and IL-6, ICAM-1 and MCP-1 were also significantly increased, resulting in oxidative stress and inflammatory injury. Guanxinning Tablet and its composed extracts could significantly improve cell morphology, increase NO level, decrease ROS production, and also reduce the secretion of inflammation-related proteins IL-6 and MCP-1. Salvia miltiorrhiza and Ligusticum striatum DC. have significant synergistic effects on NO. Among them, salvianolic acid B and salvianic acid A exerted the main effects, and the combined efficacy of salvianic acid A and ferulic acid was superior to that of single administration. The above results showed that Guanxinning Tablet and their active substances had the effects of improving endothelial basal function, resisting oxidative stress, and alleviating inflammatory injury, and Salvia miltiorrhiza and Ligusticum striatum DC. synergized, which may be related to their regulation of oxidative stress and inflammation and have application prospects in the treatment of atherosclerosis-related diseases.

Fucoxanthin suppresses OxLDL-induced inflammation via activation of Nrf2 and inhibition of NF-κB signaling

Objective: To explore the impact of fucoxanthin on oxidized low-density lipoprotein (OxLDL)-induced stress and inflammation in human endothelial cells and its underlying mechanisms. Methods: HUVECs were treated with OxLDL and/or fucoxanthin for a range of time points and concentrations. We evaluated the effects of fucoxanthin on OxLDL-induced HUVECs using the MTT assay, reactive oxygen species accumulation assay, ELISA, RT-PCR, immunofluorescence, and Western blotting. Results: Fucoxanthin enhanced the cell viability in a dose dependent manner after OxLDL exposure. Furthermore, fucoxanthin pretreatment significantly decreased OxLDL-induced reactive oxygen species production and prevented the activation of the nuclear factor kappa-B pathway, which led to substantial suppression of pro-inflammatory gene expressions. OxLDL-induced upregulation of interleukin-6, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, interleukin-1β, monocyte chemotactic protein-1, cyclooxygenase-1, and tumor necrosis factor-α was significantly reduced by fucoxanthin. Conclusions: Fucoxanthin can inhibit OxLDL-induced vascular inflammation and oxidative stress in HUVECs by targeting Nrf2 signaling pathways.

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.

Study on protective effect of total flavonoids from Rosa multiflora on HUVEC induced by ox-LDL / 中国中药杂志

The aim of this paper was to study the protective effect of total flavonoids from Rosa multiflora(TF-RM) on the injury of HUVEC induced by oxidized low density lipoprotein(ox-LDL). SPF male SD rats were randomly divided into blank group, simvastatin group(1.8 mg·kg~(-1)·d~(-1)) and TF-RM group(2.5 g·kg~(-1)·d~(-1)), with 10 rats in each group. They were intragastrically administered with drugs for 7 days, and then blood was collected from the abdominal aorta to prepare drug-containing serum. The HUVEC injury model was established through ox-LDL induction, and added with 15% simvastatin, 5% TF-RM, 10% TF-RM, 15% TF-RM drug-containing serum and blank serum, respectively. Reactive oxygen species(ROS) was determined by flow cytometry. Nitric oxide(NO) content was determined by nitrate reductase method. The contents of ET-1, P-selectin, E-selectin, ICAM-1, VCAM-1, IL-1β, IL-6 and TNF-α were determined by ELISA. The expression of Lox-1 protein was determined by Western blot. Compared with the blank group, ROS level in HUVEC and the contents of ET-1, P-selectin, E-selectin, ICAM-1, VCAM-1 and IL-1β in HUVEC were significantly increased(P<0.05), NO decreased significantly(P<0.01),Lox-1 protein expression increased significantly(P<0.05), and TNF-α and IL-6 showed an increasing trend. Compared with the model group, TF-RM significantly reduced ROS level in HUVEC and ET-1, P-selectin, E-selectin, ICAM-1, TNF-α, IL-1β content in supernatant(P<0.05), significantly increased NO content(P<0.01), and inhibited Lox-1 protein expression(P<0.05). VCAM-1, IL-6 contents showed a decreasing trend. Serum containing TF-RM acts on lectin-like oxidized low-density lipoprotein receptors, and exerts a protective effect on vascular endothelial cells by reducing cell oxidative damage, regulating vasoactive substances, and reducing adhesion molecules and inflammatory cascades.

Role of oxidized low density lipoprotein in the diagnosis of atherosclerotic cardiovascular disease / 中华检验医学杂志

As a critical factor of atherosclerosis (AS), oxidized low-density lipoprotein (oxLDL) plays a crucial role in damaging vascular endothelial cells, promoting monocyte adhesion, inducing formation of foam cells and thrombosis. Many scholars have found that oxLDL could be used as a circulating biomarker for atherosclerotic cardiovascular disease. In this manuscript, research progress on the mechanism of oxLDL in the initiation and development of atherosclerosis, as well as using oxLDL as a diagnostic biomarker of atherosclerosis related diseases were systemically reviewed.

Effect of Klotho on oxidized low density lipoprotein-induced endothelial cell senescence via activation of autophagy / 解放军医学杂志

Objective: To investigate the effect of Klotho on oxidized low density lipoprotein (ox-LDL)-induced human coronary artery endothelial cell (HCAEC) senescence via activation of autophagy. Methods: HCAECs were cultured in vitro, and divided into control group and ox-LDL group [treated with ox-LDL at different concentrations (0, 25, 50, 75, 100 μg/ml) or different durations (0, 24 h, 48 h, 72 h, 96 h) 2 h after pre-incubation with Klotho (400 pmol/L)]. Cell viability was tested by CCK-8, senescent cells were detected by SA-β-GAL staining, immunofluorescence was used to detect the intracellular expression of P53 and P16, and Western blotting was performed to quantitatively analyze the P53, P16, LC3 and P62 protein expression. HCAECs were pre-incubated separately with Klotho (400 pmol/L), rapamycin (5 μmol/L) or chloroquine (2 μmol/L) for 2 hours under normal culture condition, and then cultured under ox-LDL exposure (75 μg/ml) for 72 hours. Cell viability was tested by CCK-8, senescent cells were detected by SA-β-GAL staining, autophagic flux was assayed by adenovirus GFP-LC3, and Western blotting was performed to quantitatively analyze the P53, P16, LC3, and P62 protein expression. Results: The results of CCK-8 assay, SAKlotho β-GAL staining and Western blotting respectively showed that Klotho decreased ox-LDL-induced inhibition of HCAECs viability (P<0.05), reduced the proportion of senescent cells (P<0.05), and down-regulated the expression of aging-related proteins P53 and P16 (P<0.05). Western blotting revealed that ox-LDL decreased LC3-II/LC3-I ratio and increased P62 protein expression level in a time-dependent (P<0.05) or dose-dependent manner (P<0.05) to some extent. Furthermore, Klotho increased LC3-II/LC3-I ratio (P<0.01), decreased P62 protein expression level (P<0.01), and reduced the inhibition of autophagy by chloroquine under ox-LDL exposure (P<0.05). The results of CCK-8 assay, SA-β-GAL staining and Western blotting respectively indicated that the HCAECs viability (P<0.01), the proportion of senescent cells (P<0.01), and the protein expression levels of P53 (P<0.01) and P16 (P<0.01) were significantly alleviated in ox-LDL+Klotho group and ox-LDL+rapamycin group than those in ox-LDL group; while the HCAECs viability (P<0.01), the proportion of senescent cells (P<0.01), and the protein expression levels of P53 and P16 (P<0.05) were obviously worse in chloroquine+ox-LDL group than those in ox-LDL group. Conclusion: Klotho ameliorates ox- LDL-induced HCAEC senescence via activation of autophagy.

Relationship of the low-density lipoprotein (LDL)/high-density lipoprotein (HDL) index with antioxidant enzymes and with the oxLDL/HDL index

Introduction: The low-density lipoprotein (LDL)/high-density lipoprotein (HDL) index is a predictive factor for atherosclerosis, which is associated with oxidative modifications. Objective: To assess the association of the index with oxidative stress markers. Methods: 444 subjects were included and were clinically, anthropometrically and biochemically characterized; superoxide dismutase, glutathione peroxidase 3 (GPx3), magnesium and oxidized LDL (oxLDL) index (oxLDL/HDL) were quantified. Results: A decrease of 1.014 units in the LDL/HDL index was associated with a superoxide dismutase increase of 1 unit/mL (p = 0.030), while a decrease of 0.023 units was associated with a GPx3 increase of 1 nmol/min/mL (p < 0.0005). An increase of one unit in the index was associated with an increase of 0.831 in the oxLDL/HDL index (p < 0.05). After controlling for the effect of gender, age, smoking, obesity and insulin resistance, a reduction of 0.001 per index unit was associated with an increase of 1 µg/g of magnesium in the nails (p = 0.020). Conclusions: The LDL/HDL index shows an inverse relationship with the antioxidant status and a direct relationship with oxidation status, regardless of other cardiovascular and oxidative stress risk factors.

Effect of pomegranate polyphenols on lipids metabolism in patients with myocardial infarction: a double-blind placebo controlled trial

Background: Myocardial Infarction (MI) is a leading disease globally. Major risk factors for MI are smoking, hypertension, diabetes mellitus, reactive oxygen species (ROS), obesity, coronary artery disease (CAD) and abnormally altered blood lipid levels. It is recommended that for healthy living the risk factors for CAD and ROS should be less. Consumption of natural food supplements rich in antioxidants and polyphenols reduce the risk of MI. One herb is Pomegranate. Pomegranate is polyphenols and antioxidants rich fruit. This prompted us to find out whether the presence of antioxidants in pomegranate offers any prognostic benefits in patients with MI?.Methods: Pomegranate Extract of Whole Fruit (PEWF) was prepared as tablet of 300mg to investigate its effects in patients with MI. Total 100 participants were included in the trial. Participants were assigned to two groups of 50 each. One group received “Add On” PEWF and other got matching placebo of same colour, shape and size as comparator agent in the dose of 300mg BD for 1 month.Results: Results were compared by Z test, Chi square test and coefficient of variations. Statistical analysis proves the prognostic effect after active medication (p<0.05). Study results indicate the rejection of Null Hypothesis (H0) and acceptance of Alternative Hypothesis (H1).Conclusions: Our findings suggest that consumption of antioxidant and polyphenols rich food supplements such as PEWFs for one month reduces the risk factors for CAD.

Effects of moxibustion at 45 ℃ on blood lipoids and serum level of ox-LDL and NO in rats with hyperlipidemia / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion at different temperatures (38 ℃ and 45 ℃) on blood lipoids and serum level of oxidized low density lipoprotein (ox-LDL) and nitric oxide (NO) in rats with hyperlipidemia, and to explore the correlation between regulating blood fat and anti-oxidative stress and protection of vascular endothelium of moxibustion at 45 ℃.@*METHODS@#According to random number table, 60 SD rats were randomly divided into a normal group, a model group, a moxibustion at 38 ℃ group and a moxibustion at 45 ℃ group, 15 rats in each group. The rats in the normal group received no treatment; the rats in the remaining three groups were fed with high-fat diet for 8 weeks to prepare rat models of hyperlipidemia. After successful modeling, the rats in the model group received no treatment; the rats in the moxibustion at 38 ℃ group and moxibustion at 45 ℃ group were treated with moxibustion at "Shenque" (CV 8) and "Zusanli" (ST 36), and the temperature was controlled at (38±1) ℃ and (45±1) ℃, respectively. The moxibustion was given for 10 min at each acupoint, once every two days, and totally 4-week treatment was given. After treatment, the levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by using biochemical colorimetric method; the levels of ox-LDL and NO were measured by using ELISA method.@*RESULTS@#① Compared with the normal group, the levels of TC, TG and LDL-C were significantly increased in the model group (all 0.05). ② Compared with the normal group, the level of ox-LDL was increased but that of NO was decreased in the model group (both <0.01); compared with the model group and moxibustion at 38 ℃ group, the level of ox-LDL was decreased but that of NO was increased in the moxibustion at 45 ℃ group (<0.01, <0.05); compared with the model group, the level of ox-LDL was decreased but that of NO was increased in the moxibustion at 38 ℃ group (both <0.05).@*CONCLUSION@#Moxibustion at 45 ℃ has regulating effects on blood lipid in rats with hyperlipidemia, which can regulate blood lipid through various ways, such as anti-oxidative stress and protection of vascular endothelium.

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.

Metformin improved oxidized low-density lipoprotein-impaired mitochondrial function and increased glucose uptake involving Akt-AS160 pathway in raw264.7 macrophages / 中华医学杂志(英文版)

Background@#Macrophage accumulation in the vascular wall is a hallmark of atherosclerosis. Studies showed that shifting of oxidized lipids-induced inflammatory macrophages towards an anti-inflammatory phenotype by promoting oxidative metabolism attenuated atherosclerosis progression. Therefore, this study aimed to investigate whether metformin, which has ameliorated atherosclerosis in animal models and clinical trials, modulated oxidized low-density lipoprotein (Ox-LDL) induced inflammatory status in macrophages by regulating cellular oxidative metabolism.@*Methods@#Murine raw264.7 macrophages were incubated with Ox-LDL (50 μg/mL) in the presence or absence of metformin (15 μmol/L) for 24 h. Real-time polymerase chain reaction was used to quantify the transcription of classically activated (M1) proinflammatory and alternatively activated (M2) anti-inflammatory markers and mitochondrial DNA copy numbers. Cellular reactive oxygen species (ROS) production and mitochondrial membrane potential were detected by immunofluorescence. Cellular adenosine triphosphate (ATP) synthesis, glucose uptake, and lactic acid production were measured by commercial kit and normalized to cellular lysates. Western blotting analysis was performed to detect the expression of mitochondrial fusion/fission related proteins, enzymes mediating lipid metabolism and signaling pathway of glucose transport. Differences between groups were analyzed using one-way analysis of variance.@*Results@#Metformin improved Ox-LDL-impaired anti-inflammatory phenotype in raw264.7 macrophages as shown by up-regulated transcription of anti-inflammatory markers including interleukin 10 (0.76 ± 0.04 vs. 0.94 ± 0.01, P = 0.003) and Resistin-like molecule alpha (0.67 ± 0.08 vs. 1.78 ± 0.34, P = 0.030). Conversely, Ox-LDL-diminished phosphorylation of Akt was upregulated by metformin treatment (0.47 ± 0.05 vs. 1.02 ± 0.08, P = 0.040), associated with an improvement of mitochondrial function, characterized by decreased ROS generation (2.50 ± 0.07 vs. 2.15 ± 0.04, P = 0.040), increased lipid oxidation, and elevated cellular ATP production (0.026 ± 0.001 vs. 0.035 ± 0.003, P = 0.020). Moreover, metformin-mediated Akt activation increased Akt substrate of 160 kDa (AS160) phosphorylation (0.51 ± 0.04 vs. 1.03 ± 0.03, P = 0.0041), promoted membrane translocation of glucose transporter 1, and increased glucose influx into the cells (4.78 ± 0.04 vs. 5.47 ± 0.01, P < 0.001).@*Conclusion@#This study suggested that targeting macrophage metabolism with new or existing drugs had therapeutic potential for the prevention and treatment of diabetes-accelerated atherosclerosis.

Effect of Xiexintang Containing Serum on Expression of TLR9/MyD88/NF-κB p65 Signal Pathway in Foam Cell / 中国实验方剂学杂志

Objective: The effect of Xiexintang on Toll like receptor 9 (TLR9) signaling pathway in macrophage derived foam cells was studied by in vitro cell experiments. Method: The fifty SPF male SD rats were randomly divided into low, medium and high dose groups (1.4,4.2,12.6 g·kg-1·d -1) and normal groups. Except 20 rats in the normal group, 10 rats in each group were given equal volume of pure water gavage in the blank group. After the last Administration for 7 days, serum was separated,and the serum containing drugs in the low, medium and high dose groups of Xiexintang was prepared. Oxidized low density lipoprotein (ox-LDL) was used to intervene the differentiation of RAW264.7 macrophages into foam cells. The cell foam was identified by oil red O staining. After observing the effect of drug containing serum on the proliferation of macrophage derived foam cells by methye thiazolye telrazlium(MTT) method,the serum containing 20%concentration of each drug was selected to act on the foam cell model. The expression of interleukin(IL) -1β and interferon (INF) -γ was determined by enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR (Real-time PCR). TLR9, myeloid differentiation factor 88(MyD88)and nuclear factor(NF) -κB were detected by Western blot. Result: Oil red O staining showed that the red particles were obvious after ox-LDL intervention. The foam cell model was successfully prepared. MTT results showed that there was no significant difference in cell proliferation between the high dose group of Xiexintang in the 10%~30%concentration range and the normal group serum. Follow up selection of the serum containing 20%concentration of each dose intervened the foam cells induced by ox-LDL. Compared with the normal group,the model group after ox-LDL intervention induced the high expression of TLR9,MyD88,NF-κB p65,IL-1β,INF-γ (PPκB p65,IL-1β,INF-γ(PPConclusion: Xiexintang containing serum can inhibit ox-LDL-induced RAW264.7 macrophage foaming,and its mechanism may involve regulation of TLR9/MyD88/NF-κB p65 signaling pathway and inhibition of IL-1β and INF-γ overexpression. This may be one of its mechanisms of against atherosclerosis.

Comparative Study On Effects Of Lipoproteins On Growth And Migration Characteristics Of Tamoxifen Resistant Breast Cancer Cells

@#Introduction: Resistance towards treatment is one of the challenges in breast cancer therapy. Recent studies show the link between lipoprotein with cancer resistance and progression. Clinical data indicates that oxidized low-density lipoprotein (oxLDL) and very low-density lipoprotein (VLDL) play roles the progression of breast cancer. Therefore, purpose of this study was to determine the roles of lipoproteins on migration of breast cancer cell and compare the effects of oxLDL and VLDL. Methods: Parent MCF-7 cells were purchased from ATCC, while the Tamoxifen-resistant MCF-7 (Tam-R MCF-7) was developed by pulse treatment method. Tam-R cells were treated with gradual increase in tamoxifen concentration for 72 hours in Dulbecco’s Modified Eagle’s medium (DMEM) without phenol red. Cell viability test was done to measure the fold changes of Tam-R MCF-7 cells. Migration characteristics was studied using wound healing assay. Cells were treated with 10 μg/mL of oxLDL and VLDL up to 72 hours. Results: From the cell viability test, Tam-R MCF-7 cells had 4-fold increase of resistance than parental cells. Tam-R MCF-7 had acquired resistance to Tamoxifen and achieved a clinically relevant level of resistance. Lipoproteins were found to cause morphological changes, where cells exhibited elongation and dendritic-like growth compared to control cells. Both MCF-7 parental cells and Tam-R MCF-7 cells showed higher percentage of wound closure when treated with oxLDL. In contrast, VLDL treatment caused reduction in cell migration compared to oxLDL. Conclusion: Findings suggest that oxLDL may further promote resistant breast cancer cell migration compared to VLDL.

β2GP/anti-β2GP complex inhibits oxLDL-mediated lipid accumulation and FAK activation in THP-1 macrophages / 临床检验杂志

Objective@#To investigate the effects of β2 glycoprotein Ⅰ/anti-β2 glycoprotein Ⅰ complex (β2/aβ2) on oxidized low density lipoprotein (oxLDL)-mediated lipid accumulation and focal adhesion kinase (FAK) activation in THP-1 macrophage, as well as the role of Toll-like receptor 4 (TLR4) during the process. @*Methods@#THP-1 cells were differentiated into THP-1 macrophage by PMA (100 ng/mL). THP-1 macrophages were treated with RPMI 1640 medium, oxLDL, oxLDL+β2/aβ2 or oxLDL+lipopolysaccharide (LPS). The mRNA expressions of lipid transportation molecules, ACAT1, ABCA1 and ABCG1 were detected by RT-qPCR. Intracellular total cholesterol (TC) and free cholesterol (FC) in THP-1 macrophages were evaluated by Trinder assay, then the content and proportion of intracellular cholesteryl ester (CE) were calculated. The expression and phosphorylation of FAK were detected by immune fluorescence, RT-qPCR and western blot. To evaluate the role of TLR4, THP-1 macrophages were pre-treated with or without TLR4 inhibitor TAK-242 (1 μg/mL). @*Results@#β2/aβ2 treatment significantly inhibited oxLDL-mediated lipid accumulation and FAK expression and phosphorylation in THP-1 macrophages, which could be reversed by TLR4 blockage. @*Conclusion@#β2/aβ2 inhibits the oxLDL-mediated lipid accumulation and FAK activation of THP-1 macrophage, which is related to the function of TLR4.

Quercetin attenuates Ox-LDL-induced calcification in vascular smooth muscle cells by regulating ROS-TLR4 signaling pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine whether quercetin inhibits oxidized low-density lipoprotein (Ox-LDL)-induced osteogenic differentiation and calcification of vascular smooth muscle cells (VSMCs) and understand the underlying mechanism.</p><p><b>METHODS</b>The calcification of human VSMCs following Ox-LDL treatment was assessed using alizarin red staining and by detecting ALP activity. The mRNA expressions of the bone-related genes including Msx2, BMP2 and Osterix, and the contractile proteins including SMA and SM22a were analyzed using qPCR. The effects of quercetin were investigated on OxLDL-induced VSMC calcification and changes in ALP activity, expressions of Msx2, BMP2, Osterix, SMA and SM22a, ROS levels and SOD activity. The effect of Toll like receptor 4 (TLR4) silencing mediated by siRNA transfection on cell calcification, ALP activity, gene expressions and ROS levels were investigated.</p><p><b>RESULTS</b>Ox-LDL treatment promoted VSMC calcification and up-regulated TLR4 expression. Quercetin treatment significantly attenuated Ox-LDL-induced VSMC calcification, reduced ALP activity, down-regulated the expression levels of Msx2, BMP2 and Osterix, and up-regulated the expressions of vascular smooth muscle contractile proteins SMA and SM22a. In addition, Quercetin treatment markedly increased SOD activity, reduced ROS levels and TLR4 expression in VSMCs. Silencing TLR4 expression using TLR4 siRNA also significantly decreased calcification of the VSMCs.</p><p><b>CONCLUSIONS</b>Quercetin inhibits Ox-LDL-induced VSMC calcification in VSMCs possibly by targeting the ROS/TLR4 signaling pathway.</p>

Construction of CD36 gene silencing cell lines by lentivirus-mediated RNA interference and the effect on protein expression of caveolin-1 / 生物工程学报

CD36, the major scavenger receptor, is intimately involved in the uptake of oxLDL in macrophages. To further study the function of CD36 in macrophages, we constructed CD36 gene silence cell lines (J774A.1) by lentivirus-mediated RNA interference technique, and analyzed the effect of CD36 in caveolin-1 protein expression. At first, 5 shRNA fragments were designed and synthesized according to the coding sequence (CDS) region of CD36 gene. Next, the CD36-shRNA was inserted into lentiviral vector to yield pLKO.1-CD36-shRNA plasmid. After DNA sequencing, the pLKO.1-CD36-shRNA plasmid and psiCHECK-II-CD36 were co-transfected into the 293T cells to screen the efficient CD36-shRNA. The efficient CD36-shRNA plasmid and the helper plasmid were co-transfected into the 293T cells to package the lentivirus, and then infected the J774A.1 cells. After screening by puromycin, CD36 gene silence cell lines (J774A.1) was established. Western blotting and confocal fluorescence microscopy results showed that the CD36 silencing efficiency in the gene silence cell line was 90%. Accompanied by a decrease in CD36 protein on cell surface, oxLDL binding to CD36 was significantly inhibited, indicating that the CD36 gene silence cell line is successfully established. Finally, the oxLDL stimulation and inhibitor experiments results showed that the CD36 knockdown significantly suppresses the phosphorylation of JNK and ERK, thereby inhibiting the oxLDL-induced caveolin-1 protein expression, demonstrating that CD36 modulates the caveolin-1 protein expression through the JNK/ERK-mediated signaling transduction.

The expression of oxidized low density lipoprotein and scavenger receptor that binds phospha-tidylserine and oxidized lipoprotein in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression level of oxidized low density lipoprotein (ox-LDL) and its scavenger receptor scavenger receptor that binds phosphatidylserine and oxidized lipoprotein (SRPSOX) in patients with rheumatoid arthritis (RA),and to explore the relationship between ox-LDL and disease activity.Methods The serum ox-LDL in RA patients and healthy control group were detected by enzymelinked immunosorbent assay (ELISA),as well as the fluidox-LDL in RA,osteoarthritis (OA) and inflammatory arthritis (IA).The expression of SR-PSOX in mixed cells of RA and IApatients was detected by western blot.The expression of serum ox-LDL between RA groupand the control group was analyzed by t-test and non-parametric test.The correlation of serum ox-LDL expression levels in RA patients with C-reactive protein (CRP),erythrocyte sedimentation rate (ESR) and other inflammatory factors and disease activitywas analyzed by Pearson linear regression.Results The expression of ox-LDL in the serum of RA patients was significantly higher than that of normal control group [(3 076±131) mU/ml,(2 334±84) mU/ml,t=4.242,P＜0.01].The expression of ox-LDL in synovial fluid of RA patients was significantly higher than that of the OA group [(4 963±354) mU/ml],(3 956±347) mU/ml,t=2.372,P＜0.05).The expression of SR-PSOX in synovial fluid mixed cells of RA patients was higher than that of the IA group [(4.92±0.18) vs (0.24±0.04),t=33.53,P＜0.01].The expression of ox-LDL in serum of RA patients was negatively correlated with ESR,CRP and overall disease activity DAS28 (r=-9.42,P=0.009;r=-0.35,P=0.029 7;r=0.42,P=0.008 4).The expression of ox-LDL in the serum of RA patients with moderate disease activity was significantly higher than those patients with high disease activity [(3 302±138) mU/ml vs (2 464±228) mU/ml,t=3.335,P＜0.01],however,those with low disease activity and disease remission had higher serum ox-LDL expression but without statistical significant differences.After treated with anti-rheumatic drugs (DMARDs),serum ox-LDL of RA patients had a trend of slight increasing butwithout sign-ificant difference.The ox-LDL/LDL-C or ox-LDL/HDL-C was negatively not correlated with disease activity score in 28 (DAS28),ESR,CRP.Conclusion In RA patients,the expression of ox-LDL in the serum and synovial fluid is high and the SR-PSOX expressionin synovial fluid is also high.The serum ox-LDL levels are negatively correlated with ESR,CRP and DAS28,which are related to disease activity of RA.These findings suggest that the ox-LDL and the receptor SR-PSOX may play a role in RA pathogenesis,but needs further study.

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.