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ObjectiveTo investigate the clinical efficacy of Huangqi injection combined with Buzhong Yiqi acupuncture in the treatment of chronic fatigue syndrome （CFS） with Qi deficiency and its effects on TCM syndromes， fatigue symptoms， serum superoxide dismutase （SOD）， malondialdehyde （MDA）， and oxidized low-density lipoprotein （ox-LDL） levels. MethodA total of 200 patients with CFS of Qi deficiency were randomly divided into a control group （100 cases） and an observation group （100 cases）. The control group was treated with vitamin B compounds， and the observation group was treated with Huangqi injection combined with Buzhong Yiqi acupuncture for two weeks. The scores of TCM syndromes， fatigue symptoms， levels of serum SOD， MDA， and ox-LDL and the incidence of adverse reactions were observed and compared before and after treatment in two groups. ResultAfter treatment， the total effective rate of the control group was 54.34% （50/92）， while that of the observation group was 88.54% （85/96）. The total effective rate of the observation group was higher than that of the control group （χ2=27.13，P<0.05）. Compared with those in the two groups before treatment， scores of fatigue self-assessment scale （FSAS）， physical fatigue and mental fatigue， and sleep/rest response scores of fatigue in the two groups after treatment were significantly decreased （P<0.05）. After treatment， scores of FSAS， physical fatigue and mental fatigue， and sleep/rest response scores of fatigue in the observation group were significantly decreased compared with those in the control group （P<0.05）. Compared with those in the two groups before treatment， TCM syndrome scores in the two groups after treatment were significantly decreased （P<0.05）. After treatment， TCM syndrome scores in the observation group were significantly decreased compared with those in the control group （P<0.05）. Compared with those in the two groups before treatment， MDA levels in the two groups were significantly decreased （P<0.05）， ox-LDL levels in the observation group were significantly decreased （P<0.05）， and SOD levels were significantly increased （P<0.05）. After treatment， compared with those in the control group， the serum MDA and ox-LDL levels in the observation group were significantly decreased （P<0.05）， and the serum SOD was significantly increased （P<0.05）. No serious adverse events or adverse reactions occurred during this clinical trial. ConclusionHuangqi injection combined with Buzhong Yiqi acupuncture has a good clinical curative effect in the treatment of CFS with Qi deficiency， which can effectively improve the fatigue symptoms of patients， increase the level of SOD， and reduce the level of serum MDA and ox-LDL. It is related to the production of antioxidants， inhibiting the production of lipid peroxides， and improving the body's ability to resist oxidative stress.
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Obiective Alzheimer’s disease (AD) is a degenerative disease of the central nervous system (CNS) caused by a variety of risk factors. There are various pathological changes, but apoptosis of the neurological meridian cells is one of the most important pathological bases. Hyperlipidemia is a high-risk factor for the development of AD, which can lead to increased levels of oxidized low-density lipoprotein (ox-LDL) in brain tissues. PCSK9 is a protease closely related to lipid metabolism, but studies have shown that it may be related to the development of AD. LRP1 is abundantly expressed in neuronal cells, and it is an important transporter for the clearance of Aβ. There is now a large amount of literature confirming that PCSK9 can induce the degradation of LRP1. PI3K/AKT is an important signaling pathway in vivo, which plays an important role in apoptosis, and there is now a large amount of literature confirming that LRP1 activates the PI3K/AKT pathway, which has an anti-apoptotic effect. So can PCSK9 affect the PI3K/AKT pathway through LRP1 and thus regulate neuronal apoptosis? This deserves further investigation.The aim of this study was to explore the role of PCSK9 in mediating ox-LDL pro-apoptotic neuronal cell death and its mechanism, and then further elaborate the mechanism of hyperlipidemia leading to neurodegenerative diseases such as AD. MethodsFirstly, PC12 cells were treated with different concentrations of ox-LDL (0, 25, 50, 75 and 100 mg/L) for 24 h. Oil red O staining was used to detect lipid accumulation in PC12 cells, Hoechst33258 staining and flow cytometry to detect apoptosis in PC12 cells, ELISA to detect the content of Aβ secreted by PC12, Western blot to detect expression of SREBP2, PCSK9 and LRP1. Then PC12 cells were treated with 75 mg/L ox-LDL for different times (0, 6, 12, 24, 48 h), and Western blot were performed to detect the expression of SREBP2, PCSK9 and LRP1. Finally, after transfecting 100 nmol/L PCSK9 siRNA into PC12 cells for 48 h, PC12 cells were treated with 75 mg/L ox-LDL for 24 h, Hoechst33258 staining and flow cytometry to detect apoptosis rate of PC12 cells, and Western blot to detect PCSK9, LRP1, PI3K, AKT, P-PI3K , P-AKT, NF-κB, Bcl-2, Bax, Caspase-9 and Caspase-3 expression, and ELISA detected Aβ content secreted by PC12 cells. Resultsox-LDL increased lipid accumulation and promoted apoptosis and Aβ secretion in PC12 cells, as well as increasing the expression of SREBP2 and PCSK9 and decreasing the expression of LRP1 in PC12 cells. pCsk9 siRNA could be inhibited through the PI3K/AKT pathway and the NF-κB-Bcl-2/Bax-Caspase-9/3 pathway to inhibit ox-LDL-induced apoptosis in PC12 cells while increasing Aβ secretion in PC12 cells. Conclusionox-LDL plays a bidirectional regulatory role in ox-LDL-induced apoptosis of PC12 cells by inducing an increase in PCSK9 expression and a decrease in LRP1 expression in PC12 cells, which in turn affects different signaling pathways downstream.
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Atherosclerosis could be induced by multiple factors, including hypertension, hyperlipidemia, and smoking, and its pathogenesis has not been fully elucidated. MicroRNAs have been shown to possess great anti-atherosclerotic potential, but the precise function of miR-92a-3p in atherosclerosis and its potential molecular mechanism have not been well clarified. Flow cytometry assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol-3-ium bromide (MTT) assay were performed to evaluate effects of oxidized low-density lipoprotein (ox-LDL) on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs), respectively. Malondialdehyde and superoxide dismutase levels in cell lysate were assessed with biochemical kits. The expression levels of miR-92a-3p and Sirtuin6 (SIRT6) in HUVECs exposed to ox-LDL were estimated by real-time quantitative polymerase chain reaction (RT-qPCR). In addition, the protein levels of SIRT6, c-Jun N-terminal kinase (JNK), phosphorylation JNK (p-JNK), p38 mitogen activated protein kinase (p38 MAPK), and phosphorylation p38 MAPK (p-p38 MAPK) were measured by western blot assays. The relationship between miR-92a-3p and SIRT6 was confirmed by dual-luciferase reporter assay. Ox-LDL induced apoptosis and oxidative stress in HUVECs in concentration- and time-dependent manners. Conversely, miR-92a-3p silencing inhibited apoptosis and SIRT6 expression in HUVECs. The overexpression of miR-92a-3p enhanced apoptosis and phosphorylation levels of JNK and p38 MAPK as well as inhibited proliferation in ox-LDL-induced HUVECs. In addition, SIRT6 was a target of miR-92a-3p. miR-92a-3p negatively regulated SIRT6 expression in ox-LDL-induced HUVECs to activate MAPK signaling pathway in vitro. In summary, miR-92a-3p promoted HUVECs apoptosis and suppressed proliferation in ox-LDL-induced HUVECs by targeting SIRT6 expression and activating MAPK signaling pathway.
Subject(s)
Humans , MAP Kinase Signaling System , Apoptosis , Sirtuins/genetics , MicroRNAs/genetics , Human Umbilical Vein Endothelial Cells , Lipoproteins, LDL/pharmacologyABSTRACT
@#Objective To explore the effects of ox-LDL on the biosynthesis of resolving D2 (RvD2) by human periphery blood mononuclear cells (PBMCs)-derived macrophages in ischemic stroke (IS).Methods Matured macrophages were derived from PBMCs of ischemic stroke patients and healthy controls,and were grouped to vehicle group and oxidized low-density lipoprotein (ox-LDL) group.We compared the level of RvD2,15-lipoxygenase (15-LOX) and 5-lipoxygenase (5-LOX),as well as G-protein coupled receptor 18 (GPR18).between groups,and determined the formation of foam cells.Results Compared with vehicle group,the level of RvD2(P<0.001),15-LOX and GPR18(P<0.05) in ox-LDL stimulated group was significantly increased,however,no significant difference in the expression of 5-LOX between ox-LDL and vehicle group was observed.After ox-LDL stimulation,compared with healthy control,elevation of RvD2 synthesis in ischemic stroke group was significantly lower（P<0.05）,and foam cell formation was also more severe in macrophages of stroke patients.Conclusion Ox-LDL stimulates biosynthesis of RvD2 by human macrophage,perhaps through enhancing 15-LOX expression.Moreover,function of increasing RvD2 synthesis upon ox-LDL stimulation is impaired in stroke patients.
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Oxidative low-density lipoprotein (ox-LDL)-induced endothelial cell injury is a key contributor toatherosclerosis development. However, the role and mechanism of long noncoding RNA X-inactive specifictranscript (XIST) in atherosclerosis remain largely unknown. The ox-LDL-induced human umbilical veinendothelial cells (HUVECs) injury was analyzed by cell viability, apoptosis, inflammatory cytokines secretionand oxidative stress. The expression levels of XIST, microRNA-204-5p (miR-204-5p) and toll-like receptor 4(TLR4) were detected by quantitative real-time polymerase chain reaction and western blot, respectively. Thetarget interaction between miR-204-5p and XIST or TLR4 was explored by bioinformatics analysis, luciferaseassay and RNA immunoprecipitation. The expression of XIST was enhanced in ox-LDL-treatedHUVECs. Knockdown of XIST attenuated ox-LDL-induced viability inhibition, apoptosis production,inflammatory response and oxidative stress in HUVECs. XIST was validated as a sponge of miR-204-5pand TLR4 acted as a target of miR-204-5p. Knockdown of miR-204-5p reversed silence of XISTmediated suppressive role in ox-LDL-induced injury. TLR4 alleviated miR-204-5p-mediated inhibitiveeffect on ox-LDL-induced injury. Moreover, XIST could regulate TLR4 expression by spongingmiR-204-5p. In conclusion, silence of XIST displayed a protective role in ox-LDL-induced injury inHUVECs by regulating miR-204-5p/TLR4 axis, providing a novel mechanism for understanding thepathogenesis of atherosclerosis.
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This research aimed to explore the molecular mechanism of microRNA (miR)-106b in cell apoptosis of atherosclerosis (AS). Human aortic endothelial cells (HAECs) were divided into control group, oxidized-low-density lipoproteins (ox-LDL) group, miR-106b NC+ox-LDL group, miR-106b mimics+ox-LDL group, miR-106b mimics+PTEN+ox-LDL group, and miR-106b mimics+empty+ox-LDL group. Real-time fluorescence quantitative polymerase chain reaction, cholecystokinin, TdT-mediated biotinylated nick end-labeling assay, luciferase reporter gene assay, and flow cytometry analysis were performed to determine the morphology, proliferation, and apoptosis in HSECs. Moreover, the levels of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), Bcl-2, p-P13K, and p-AKT in HAECs were detected by western blot. MiR-106b was down-regulated in ox-LDL-induced HAECs. PTEN was the target gene of miR-106b-5p. Overexpression of PTEN inhibited the anti-apoptotic effect of miR-106b. Compared with the control group, the proportion and number of HAECs apoptosis and Bax, caspase-3, and caspase-9 expression in ox-LDL and miR-106b mimics+PTEN+ox-LDL groups were significantly increased (all P<0.05). Moreover, the activity of HAECs and Bcl-2 were decreased significantly (all P<0.05). Overexpression of miR-106b in ox-LDL-induced AS inhibited endothelial cell apoptosis. Furthermore, miR-106b might activate the PI3K/AKT pathway by down-regulating the expression of PTEN in ox-LDL-induced HAECs.
Subject(s)
Humans , Apoptosis , MicroRNAs/genetics , Endothelial Cells/metabolism , Atherosclerosis/metabolism , Lipoproteins, LDL/genetics , Signal Transduction , Up-Regulation , Cell Proliferation , Real-Time Polymerase Chain Reaction , Fluorescence , Lipoproteins, LDL/metabolismABSTRACT
Aim To study the effects of trimetazidine (TMZ)on theformationof neutrophil extracellular traps (NETs) induced by oxidized low-density lipoprotein (ox-LDL) in vitro and its relationship associated with cell autophagy. Methods The bone marrow neutrophils of mice were extracted by density gradient centrifugation,and NETs induction model was established using ox-LDL. Furthermore, TMZ, autophagy inhibitor LY294002 and autophagy inducer Rapamycin were added to disturb the induction of NETs induced by ox-LDL. The production of NETs marker MPO-DNA and the content of cfDNA/nets in the supernatantwere observed. Meanwhile, Western blot was employed to determine the protein expression of myeloperoxidase (MPO),beclin-l and LC3b in neutrophils. Results Treatment of ox-LDL to adhered neutrophils could lead to the formation of extracellular MPO-DNA and cfDNA/nets generation in a time-and concentration-dependent manner. The protein expression of LC3b,beclin-1 and MPO in neutrophils was up-regulated after treatedwith ox-LDL. TMZ pretreatment significantly inhibited NETs release and reduced the protein expression of LC3b,beclin-1 and MPO in neutrophils,which could be simulated by PI3K/AKT pathway blocker LY294002, while AKT/mTOR inhibitor rapamycin preconditioning did the opposite. Conclusions Ox-LDL-induces the formation of NETs in a time-and concentration-dependent manner. TMZ could down-regulate the level of autophagy and neutrophils, inhibit the induction of ox-LDL on NETs and reduce the release of NETs.
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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.
Subject(s)
Animals , Rats , Hyperlipidemias , Lipoproteins, LDL , Moxibustion , Nitric Oxide , Rats, Sprague-DawleyABSTRACT
Objective@#To investigate the effects of miR-27b targeting ten-eleven translocation methylcytosine dioxygenase 2 (TET2) on oxidized low-density lipoprotein (ox-LDL) induced inflammatory responses and apoptosis in endothelial cells.@*Methods@#Double luciferase reporter gene analysis verified the targeting effect of miR-27b on TET2. Human umbilical vein endothelial cells (HUVECs) were induced by ox-LDL in vitro. Eight groups were set up including control group, ox-LDL group, ox-LDL+ anti-miR-con group, ox-LDL+ anti-miR-27b group, ox-LDL+ pcDNA group, ox-LDL+ pcDNA-TET2 group, anti-miR-27b+ si-con group and anti-miR-27b+ si-TET2 group. qRT-PCR was used to detect the expression of miR-27b and TET2 at mRNA level. Cell viability was detected by MTT assay. Cell apoptosis rate was detected by flow cytometry. Western blot was used to detect the expression of TET2, Cyclin D1 and caspase-3 at protein level.@*Results@#TET2 was the target gene of miR-27b. TET2 expression could be negatively regulated by miR-27b. ox-LDL increased the expression of miR-27b and reduced the expression of TET2 in HUVECs. The secretion of inflammatory factors and apoptosis rates of HUVECs in the control, ox-LDL+ anti-miR-27b, ox-LDL+ pcDNA-TET2 and anti-miR-27b+ si-con groups were significantly lower than those in the ox-LDL, ox-LDL+ anti-miR-con, ox-LDL+ pcDNA and anti-miR-27b+ si-TET2 groups, respectively (P<0.05).@*Conclusions@#miR-27b promoted ox-LDL-induced inflammatory responses and apoptosis in endothelial cells through down-regulating the expression of TET2.
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Objective@#To investigate the role of lectin-like Ox-LDL receptor-1( LOX-1) in the activation and oxidative stress of cultured human umbilical vein endothelial cell(HUVEC) after human cytomegalovirus(HCMV) infection.@*Methods@#HUVEC were divided into four groups: HCMV, Control, Carrageenan, and HCMV+ Carrageenan. After HCMV AD169 infection, the supernatant of the culture was extracted, and cells were lysed. The levels of LOX-1 mRNA, intercellular adhesion molecule-1(ICAM-1) mRNA and vascular cellular adhesion molecule-1(VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxidum(NO) of the supernatant was detected by nitrate reductase method accordingly.@*Results@#24 h after infection, the mRNA expression of LOX-1, ICAM-1 and VCAM-1 in HUVEC of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly. The mRNA expression of LOX-1, ICAM-1 and VCAM-1 and the quantity of NO decreased after adding the LOX-1 inhibitor carrageenan. There was significant difference between groups(P<0.05).@*Conclusions@#HCMV may increase the mRNA expression of ICAM-1 and VCAM-1 and quantity of NO by upregulating the mRNA expresion of LOX-1, which may contribute to the formation of a therosclerosis(AS).
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Objective To clarify the role of human β-defensin2 ( hBD2) on preventing oxidized low-density lipoprotein (OX-LDL) induced human leukemic monocyte (THP-1) foaming.Methods The monocyte foaming model was established using THP-1 cell induced by OX-LDL and the model was identified by oil red staining.The hBD2 was overexpressed on THP-1 cells by using lentivirus system and the effect of hBD 2 overexpression on THP-1 cell foaming induced by OX-LDL was detected.The levels of inflammatory factors including tumor necrosis factor (TNF)-α, interleukin (IL)-1βand IL-6 in cell supernatant of each group were detected by enzyme-linked immunosorbent assay ( ELISA).Differences between the groups were compared by using the t test.Results The gene transfection efficiency of the cells was close to 100%at 72 h after infection. The hBD2 protein levels were 0.122 ±0.024 in the control group, 0.123 ±0.022 in Lv-control infection group and 0.981 ±0.183 in Lv-hBD2 infection group; and the level in control group was statistically higher than that in hBD-2 infection group (t＝-3.175, P＝0.007).The relative levels of hBD2 mRNA at 72 h after virus infection were 0.131 ±0.021 in control group, 0.128 ±0.022 in Lv-control group and 1.001 ±0.105 in Lv-hBD2 infection group; and the level in control group was statistically higher than that in hBD-2 infection group (t＝-7.213, P＝0.003).The results of oil red staining showed that OX-LDL inducing THP-1 cells for 72 h could significantly induce lipid accumulation in cells.Overexpression of hBD2 could effectively inhibit lipid accumulation in THP-1 cells induced by OX-LDL.The expression of hBD2 mRNA in THP-1 group was significantly higher than that in THP-1+OX-LDL group (t＝3.237, P＝0.004); and the difference was also significant when comparing THP-1+Lv-hBD2+OX-LDL group with THP-1+OX-LDL group (t＝-6.021, P＝0.003).The level of hBD2 protein in THP-1 group was significantly higher than that in THP-1+OX-LDL group (t＝0.314, P＝0.006); and the difference was also significant when comparing THP-1+Lv-hBD2+OX-LDL group with THP-1+OX-LDL group (t＝-4.061,P＝0.007).The levels of TNF-α, IL-1βand IL-6 in the supernatant of THP-1 cells induced by OX-LDL for 72 h were significantly increased compared with those in THP-1group (t＝-3.825,-2.017 and -3.551, respectively; P＝0.007, 0.004 and 0.005, respectively). The levels of TNF-α, IL-1βand IL-6 in THP-1+Lv-hBD2+OX-LDL group were significantly lower than those in THP-1+OX-LDL group ( t＝4.132, 3.681, and 2.991, respectively; P＝0.003, 0.002, and 0.007, respectively).Conclusions hBD2 can effectively inhibit THP-1 foaming induced by OX-LDL, which may be related to its inhibition of inflammatory response.
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PURPOSE: Previous study has well documented the anti-apoptotic effects of miR-590 on oxidized low-density lipoprotein (ox-LDL)-treated endothelial cells (ECs). However, the mechanism underlying the anti-apoptotic effects of miR-590 in ox-LDL-treated ECs remains to be further addressed. MATERIALS AND METHODS: ApoE(−/−) mice fed with a high-fat diet (HFD) and human aortic endothelial cells (HAECs) treated with ox-LDL were used as in vivo and in vitro models of atherosclerosis. The expressions of miR-590 and toll-like receptor 4 (TLR4) were detected by quantitative real-time PCR and Western blot, respectively. Atherosclerotic lesion analysis was performed using Evans blue and hematoxylin-eosin staining. Cell proliferation was assessed by MTT assay. Apoptosis was examined using flow cytometry analysis and Western blot analysis of Cleaved poly (ADP-ribose) polymerase (PARP) and Cleaved Caspase-3 levels. The effect of miR-590 on TLR4/nuclear factor kappa B (NF-κB) pathway was evaluated by Western blot. Binding between miR-590 and TLR4 was confirmed by luciferase reporter assay and Western blot. RESULTS: miR-590 was downregulated in the aorta tissues from HFD-fed apoE(−/−) mice and ox-LDL-treated HAECs. miR-590 overexpression inhibited atherosclerotic lesion in HFD-induced apoE(−/−) mice and promoted proliferation and inhibited apoptosis of ox-LDL-treated HAECs. Additionally, TLR4 was identified as a direct target of miR-590 in ox-LDL-treated HAECs. Moreover, anti-miR-590 reversed TLR4 knockdown-mediated promotion of cell proliferation and suppression of apoptosis in ox-LDL-treated HAECs. miR-590 overexpression suppressed the TLR4/NF-κB pathway, and inhibition of the TLR4/NF-κB pathway promoted cell proliferation and impeded apoptosis in ox-LDL-treated HAECs. CONCLUSION: miR-590 promoted proliferation and blocked ox-LDL-induced apoptosis in HAECs through inhibition of the TLR4/NF-κB pathway.
Subject(s)
Animals , Humans , Mice , Aorta , Apoptosis , Atherosclerosis , Blotting, Western , Caspase 3 , Cell Proliferation , Diet, High-Fat , Endothelial Cells , Evans Blue , Flow Cytometry , In Vitro Techniques , Lipoproteins , Luciferases , Real-Time Polymerase Chain Reaction , Toll-Like Receptor 4ABSTRACT
Objective To investigate the effects of puerarin on the expression of human umbilical vein endothelial cells (HUVECs) tissue factor (TF) and tissue factor pathway inhibitor (TFPI) induced by oxidized low-density lipoprotein (ox-LDL).Methods After HUVECs were incubated with different concentrations of puerarin and 50 mg/L ox-LDL,the expression of TF and TFPI mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blot respectively.Results Compared with control,treatment with ox-LDL caused the augment of TF mRNA and protein expression (P<0.01),and the decrease of TFPI mRNA and protein expression.However,50,100,and 200 μmol/L puerarin blunted the augment of TF mRNA and protein expression and weakened the inhibition of TFPI mRNA and protein expression induced by ox-LDL(P<0.01).Conclusions Puerarin reduces HUVECs TF and TFPI mRNA and protein induced by ox-LDL.
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Objective:To investigate the role of scavenger receptor BⅡ(SR-BⅡ) in the oxidized low density lipid(ox-LDL) induced foam cells formation promoted by porphyromonas gingivalis(P.g).Methods:Peritoneal macrophages were stimulated with P.g and ox-LDL,and then foam cells formation were checked.The expression of SR-BⅡ was detected by Western blot and real-time PCR.Next,siRNA targeting SR-BⅡ was used to detect the change of foam cells formation.Results:After being stimulated with P.g and ox-LDL,the foam cells formation was significantly increased.During the process of foam cells formation,P.g infection increased the expression of SR-BⅡ.And the knockdown of SR-BⅡ by siRNA significantly reduced the foam cells formation.Conclusion:P.g infection can increase the expression of SR-BⅡ and the regulation of SR-BⅡ expression can change the foam cells formation.
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OBJECTIVE:To observe clinical efficacy and safety of Mailuoning injection in adjunctive treatment of acute cere-bral infarction. METHODS:A total of 65 patients with acute cerebral infarction selected from neurology department of our hospital were divided into control group(32 cases)and observation group(33 cases)according to random number table. Control group was given conventional treatment. Observation group was additionally given Mailuoning injection 10 mL added into 0.9% sodium chlo-ride injection 250 mL intrnrenously,ivgtt,qd,on the basis of control group. Both group were treated for 15 d. Clinical efficacy as well as serum levels of ox-LDL,BNP and MMP-9,NIHSS score before and after treatment,the occurrence of ADR were com-pared between 2 groups. RESULTS:The response rate of observation group was 90.91%,which was significantly higher than 65.63%,with statistical significance(P0.05). After treatment,serum levels of ox-LDL,BNP and MMP-9,NIHSS score in 2 groups were all decreased significantly,and the observation group was significantly lower than the control group,with statis-tical significance (P0.05). CONCLU-SIONS:Mailuoning injection has significant therapeutic efficacy for acute cerebral infarction,can significantly reduce serum levels of ox-LDL,BNP and MMP-9,promotes neurological function and the recovery of patients with cerebral infarction with good safety.
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To observe the functions of Gualou Xiebai Banxia decoction（GXBD） on regulating lipid metabolism, anti-oxidation, and interposing ox-LDL/Lox-1 pathway, and to explore its anti-atherosclerosis (AS) mechanisms. AS models were established by using 42 Apo-E-/- male mice with high fat diet. AS model mice were randomly divided into the model group, simvastatin group, and GXBD high and low dose groups. C57BL/6J male mice were used as the normal control group, n=10 and the treatment lasted for 8 weeks. The levels of TC, TG, LDL-C, HDL-C, SOD, MDA, GSH-px, and ox-LDL in blood serum were tested 24 h after the last administration. The changes of aortic tissues structure were observed by HE staining; the expression levels of Lox-1 protein and the expression levels of mRNA were detected by Western blot and PCR respectively.Results showed that the blood lipid levels and MDA, ox-LDL levels in blood serum of model group were significantly higher than those in the normal control group, but SOD, GSH-px levels were significantly lower than those in the normal control group, and the Lox-1 protein and mRNA expression levels were also significantly higher than those in the control group(P<0.05), namely aortic atherosclerosis lesions were obvious in model group.The levels of blood lipid and MDA, ox-LDL of GXBD high and low dose groups and simvastatin group were significantly lower than those in model group, while SOD, GSH-px levels were significantly higher than those in model group, and Lox-1 protein and mRNA expression levels were significantly lower than those in model group(P<0.05), namely the aortic atherosclerosis lesions were significantly relieved. The above results indicated that GXBD was capable of modulating blood lipid, anti-oxidation, and inhibiting the expression of Lox-1, and interposing ox-LDL/Lox-1 pathway in the AS model Apo-E-/- mice, which may be one of the mechanisms of anti-atherosclerosis.
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The protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against ox-LDL-induced damage to human umbilical vein endothelial cells (HUVECs) was investigated by MTT assay, and the components were identified by using UPLC-Q-TOF-MS. According to the study, ethyl acetate extract and n-butanol extract could increase cell viability (P<0.01), while petroleum ether extract had no influence, and water extract could even inhibit the cell viability to some degree. Moreover, 32 compounds in four polar fractions were analyzed, including 31 quinones and their glycosides, and one rubiprasins C. Petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract contained 23, 32, 26, 15 compounds, respectively. According to cell experiments in vitro, active fractions were ethyl acetate extract and n-butanol extract. The results could provide scientific references for further studies on effective material basic of cRRR, and lay a foundation for studies on the relationship between efficacies and materials.
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Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.
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Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.
ABSTRACT
Objective To investigate the protective effect of curcumin on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL) and its possible mechanism.Methods Cultivated HUVECs were divided into six groups: control group, ox-LDL group, ox-LDL plus endoplasmic reticulum stress(ERS) inhibitor PBA group,curcumin group, ox-LDL plus curcumin group,ox-LDL plus curcumin plus PI3K inhibitor LY294002 group.Cell viabilities were evaluated by CCK-8 assays.The proportions of apoptotic cells were assessed by flow cytometry.The translocation of activating transcription factor 6 (ATF6) observed by laser confocal microscopy.Western blot was used to determine the expression of the ERS associated proteins:glucose-regulated protein 78(GRP78), protein kinase-like ER kinase(PERK), inositol-requiring kinase1(IRE-1) and the related pathways protein: LOX-1, AKT and phophorylated AKT.Results Compared with control group,increasedthe proportions of apoptotic cells(P<0.01),enhanced the expressions of ERS related proteins(P<0.01),promoted the transfer of ATF6 into the nucleus,as well as increased the expression of LOX-1(P<0.01)and decreased the expression of p-AKT(P<0.01) in the ox-LDL group;Compared with ox-LDL group,PBA inhibited ox-LDL-induced HUVECs apoptosis(P<0.01),curcumin inhibited ox-LDL-induced the expression of ERS associated protein and LOX-1(P<0.01), the nuclear translocation of ATF6, the apoptosis of HUVECs (P<0.01), and it also increased ox-LDL-induced down-regulation of p-AKT expression (P<0.01);LY294002 partially attenuated the inhibitory effect of curcumin on ERstress-related protein expression induced by ox-LDL(P<0.05).ConclusionsCurcumin can reduce ox-LDL induced apoptosis of HUVECs, its mechanism may be through the inhibition of LOX-1 expression and activation of AKT pathway to reduce ERS in cell.