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ObjectiveTo investigate the mechanism of Renshentang， recorded in Synopsis of Golden Chamber， in the treatment of atherosclerosis （AS） based on the autophagic effect of transient receptor potential vanilloid subtype 1 （TRPV1） on arterial smooth muscle. MethodFourteen SPF-grade 8-week-old male C57BL/6J mice were assigned to the normal group and 70 8-week-old apolipoprotein E knockout （ApoE-/-） mice were assigned to the experimental group. The AS model was induced by a high-fat diet in the mice in the experimental group for eight weeks. The model mice were then randomly divided into model group， low-， medium-， and high-dose Renshentang groups （2.715， 5.43， and 10.68 g·kg-1·d-1）， and simvastatin group （0.02 g·kg-1·d-1）. Drug treatment lasted eight weeks. Serum was taken and serum total cholesterol （CHO）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） levels were measured by assay kits to observe the changes in lipid levels in mice. The aorta was stained with hematoxylin-eosin （HE） to observe the overall pathology of the aortic root and oil red O staining was used to detect the lipid deposition in the aortic plaque and calculate the percentage of the aortic root area to the lumen area. The protein expression of TRPV1， adenylate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， autophagy effector-1 （Beclin-1）， and microtubule-associated protein 1 light chain 3 （LC3Ⅱ） in mouse aortic tissues was determined by Western blot. ResultCompared with the normal group， the model group showed increased serum CHO， TG， and LDL-C levels， decreased HDL-C， and increased aortic root plaque area （P<0.01）. Compared with the model group， the Renshentang groups showed decreased levels of CHO， TG， and LDL-C in serum （P<0.05， P<0.01）， especially in the low- and medium-dose Renshentang groups （P<0.01）. Compared with the normal group， the simvastatin group and the Renshentang groups showed reduced aortic root plaque area （P<0.05）， especially in the high-dose Renshentang group （P<0.01）. Compared with the normal group， the model group showed decreased relative expression levels of TRPV1， p-AMPK/AMPK， Beclin-1， and LC3Ⅱ/LC3Ⅰ（P<0.05， P<0.01）. Compared with the model group， the medium- and high-dose Renshentang groups showed increased relative expression levels of TRPV1， p-AMPK/AMPK， Beclin-1， and LC3Ⅱ/LC3Ⅰ（P<0.05，P<0.01）. ConclusionThe anti-AS effect of Renshentang recorded in Synopsis of Golden Chamber may be achieved by up-regulating TRPV1 expression to restore the level of autophagy mediated by AMPK.
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Objective:To explore the correlation between the efficacy of <italic>Usnea diffracta</italic> in treating atherosclerosis （AS） and the altered microbial flora in rat ileum based on the interior-exterior relationship between heart and small intestine in traditional Chinese medicine （TCM）. Method:Forty-eight SD rats were randomized into a normal group （<italic>n</italic>=8） and a modeling group （<italic>n</italic>=40）. The AS model was established with high-fat diet combined with intraperitoneal injection of vitamin D<sub>3</sub>. The successfully modeled rats were further randomly divided into the model group， positive control （simvastatin， 4 mg·kg<sup>-1</sup>） group， and low- （0.7 g·kg<sup>-1</sup>）， medium- （1.4 g·kg<sup>-1</sup>）， and high-dose （2.8 g·kg<sup>-1</sup>） <italic>U. diffracta</italic> ethanol extract groups， with eight rats in each group. After four weeks of intervention， the blood， aorta， ileum， and ileum content of rats in each group were collected. The levels of serum lipopolysaccharide （LPS）， tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） and interleukin 6 （IL-6） were measured by enzyme-linked immunosorbent assay （ELISA）， and the pathological changes in rat thoracic aorta was detected by hematoxylin-eosin （HE） staining. Western blot was conducted to determine the protein expression levels of tight junction protein zonula occluden （ZO-1） and Occludin in rat ileum， and the high-throughput 16S rRNA sequencing technology was employed to detect changes in microbial diversity and abundance in rat ileum of each group. Result:Compared with the normal group， the model group exhibited obvious aortic plaque deposition， increased LPS， TNF-<italic>α</italic>， and IL-6 levels （<italic>P</italic><0.01）， but decreased ZO-1 and Occludin protein expression （<italic>P</italic><0.01）. The comparison with the model group revealed that <italic>U. diffracta</italic> significantly ameliorated the aortic plaque deposition of model rats， lowered serum LPS， TNF-<italic>α</italic>， and IL-6 levels （<italic>P</italic><0.05， <italic>P</italic><0.01）， and up-regulated ZO-1 and Occludin protein expression （<italic>P</italic><0.05， <italic>P</italic><0.01）. The abundance of Proteobacteria and Bacteroidetes in the model group changed significantly in contrast to that in the normal group， and the Bacteroidetes/Firmicutes（B/F） value declined （<italic>P</italic><0.05）. Alpha and Beta diversity analysis indicated higher total number of intestinal flora species in the model group， but lower richness and uneven distribution （<italic>P</italic><0.05， <italic>P</italic><0.01）， with a large number of pathogenic bacteria enriched. The ethanol extract of <italic>U. diffracta</italic> significantly increased the B/F value， corrected the structural disorder of microbial flora in ileum， reduced pathogenic bacteria， and increased the relative abundance of probiotics. Conclusion:<italic>U. diffracta</italic> exerts the therapeutic effect against AS possibly by improving the intestinal microbial communities， strengthening the intestinal mucosal barrier function， and reducing the serum LPS and inflammatory factors.
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Objective:To investigate the effects of Di'ao Xinxuekang （DXXK） on NLRP3 inflammasome in mouse RAW264.7 macrophages and thoracic aorta of rats with atherosclerosis （AS）， so as to explore its anti-AS mechanism. Method:RAW264.7 cells were stimulated with oxidized low density lipoprotein （ox-LDL） and then intervened with MCC950 and DXXK. The contents of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） were determined by enzyme linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of Nod-like receptor protein 3 （NLRP3）， inflammasome adaptor protein apoptosis-associated speck-like protein containing CARD （ASC）， and cysteine-dependent aspartate-directed protease-1 （Caspase-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blotting. Sixty male SD rats were randomly divided into the normal group， model group， atorvastatin group （2.0 mg·kg<sup>-1</sup>）， as well as high-， medium-， and low-dose （100， 30， and 10 mg·kg<sup>-1</sup>） DXKK groups， with 10 rats in each group. The rats were exposed to the high-fat diet and vitamin D<sub>2</sub> for inducing AS. The blood lipid level was measured using an automatic biochemical analyzer， followed by the calculation of AS index （AI）. The contents of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> were determined by ELISA， and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in thoracic aorta were assayed by Real-time PCR and Western blotting. HE staining and Sirius red staining were conducted to observe the pathomorphological changes in the abdominal aorta and aortic sinus. Result:Compared with the normal group， the model group exhibited significantly increased TNF-<italic>α</italic> and IL-1<italic>β</italic> contents and up-regulated NLRP3， ASC， and Caspase-1 mRNA and protein expression in RAW264.7 cells （<italic>P</italic><0.01）. The above indexes in each drug administration group were significantly reduced in contrast to those in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. The comparison with the model group showed that cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and AI in each DXXK group significantly declined， while the high-density lipoprotein cholesterol （HDL-C） was significantly elevated （<italic>P</italic><0.05， <italic>P</italic><0.01）. The levels of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in the thoracic aorta were decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Abdominal aortic lesions and fibrous hyperplasia of aortic sinus were significantly improved. Conclusion:DXXK has a significant anti-AS effect， which is possibly related to the inhibition of NLRP3 inflammasome.
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Cells in the arterial wall are constantly subjected to the shear stress generated by the blood flow. Shear stress plays a pivotal role in the formation of atherosclerosis. The endothelial cells located between the blood and the vessel wall have a unique response to the shear stress of the blood flow， which can convert mechanical stimulation into intracellular signals， thereby affecting the pathological process of atherosclerosis. Endothelial function is not only regulated by hormones， growth factors and other biochemical substances， but also affected by mechanical forces such as blood flow shear stress. Physiologically， shear stress can play an anti-atherosclerotic role in maintaining the homeostasis of endothelial cells. Pathological shear stress will lead to endothelial dysfunction and promote the progression of atherosclerosis. Under the mediation of different shear stress， the endothelial function can be regulated through epigenetic pathways or mechanically sensitive cation channels. Therefore， it is necessary to understand how various signal transduction pathways are affected by pathological shear stress， so as to cause endothelial dysfunction and atherosclerosis. Traditional Chinese medicine（TCM） has been increasingly recognized for its curative effect in treating atherosclerosis， with the advantages of few side effects， multiple targets and multiple mechanisms. In recent years， the understanding of the anti-atherosclerosis mechanism of TCM mediated by shear stress has gradually deepened. This review will take endothelial function as the breakthrough point， systematically sort out the influence of shear stress on the pathological process of atherosclerosis and the related molecular mechanisms. Meanwhile， it is the first time to summarize the latest research progress of Chinese medicine against shear stress damage by sorting out the existing literature. This article mainly clarify the relationship between shear stress， endothelial function， atherosclerosis and TCM， in order to provide a theoretical basis for the clinical treatment and pathological mechanism of atherosclerosis.
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Objective To study the effects of abnormal blood flow on the secretion of ET-1/NO and the expression of the mRNA and the protein of ET-1, eNOS, VCAM-1, ICAM-1 and MCP-1 in human umbilical vein endothelial cells (HUVECs), so as to explore the mechanism of atherosclerosis （AS） caused by abnormal hemodynamics. MethodsThe HUVECs were divided into stress group, wall pressure group and normal group according to the different stress. The HUVECs were cultured under the corresponding stress for 24 hours and then collected. The secretion levels of NO and ET-1 were detected by enzyme method and radioimmunoassay method. The mRNA expression levels of eNOS and ET-1 were detected by qPCR. The expression levels of the mRNA and the protein of VCAM-1, ICAM-1, MCP-1 were detected by qPCR and Western blot. Results Compared with normal group, the secretion level and the mRNA expression level of ET-1 in wall pressure group increased significantly (P＜0.01), and the secretion level of NO and the mRNA expression level of eNOS in stress group also increased significantly (P＜0.01), The expressions level of the mRNA and the protein of VCAM-1, ICAM-1 and MCP-1 obviously increased in stress group and wall pressure group (P＜0.01). Conclusions Stress or wall pressure acting on HUVECs alone could lead to its dysfunction of the secretion and the expression of gene and protein. The mechanism of AS caused by abnormal blood flow was related to these dysfunction of HUVEC.
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Objective To study the effects of abnormal blood flow on the secretion of ET-1/NO and the expression of the mRNA and the protein of ET-1, eNOS, VCAM-1, ICAM-1 and MCP-1 in human umbilical vein endothelial cells (HUVECs), so as to explore the mechanism of atherosclerosis （AS） caused by abnormal hemodynamics. MethodsThe HUVECs were divided into stress group, wall pressure group and normal group according to the different stress. The HUVECs were cultured under the corresponding stress for 24 hours and then collected. The secretion levels of NO and ET-1 were detected by enzyme method and radioimmunoassay method. The mRNA expression levels of eNOS and ET-1 were detected by qPCR. The expression levels of the mRNA and the protein of VCAM-1, ICAM-1, MCP-1 were detected by qPCR and Western blot. Results Compared with normal group, the secretion level and the mRNA expression level of ET-1 in wall pressure group increased significantly (P＜0.01), and the secretion level of NO and the mRNA expression level of eNOS in stress group also increased significantly (P＜0.01), The expressions level of the mRNA and the protein of VCAM-1, ICAM-1 and MCP-1 obviously increased in stress group and wall pressure group (P＜0.01). Conclusions Stress or wall pressure acting on HUVECs alone could lead to its dysfunction of the secretion and the expression of gene and protein. The mechanism of AS caused by abnormal blood flow was related to these dysfunction of HUVEC.
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Objective: To screen the anti-atherosclerosis (AS) activity of the compounds by using THP-1-HIF-1α-HER-Luciferase high-throughput model, and to verify the anti-AS function of the effective compounds. Methods: THP-1-HIF-1α-HER-Luciferase cells were pretreated with different concentrations of compounds (1, 10, and 100 μg/mL) for 2 h, then cultured under hypoxia for 24 h. Luciferase activity of cells was detected and compounds with anti-AS activity were screened by luciferase activity evaluation. THP-1 and U937 cells were pretreated with effective compounds, and then induced for 24 h by oxidized low density lipoprotein (OX-LDL). The formation of foam cells was observed by oil red staining. The mRNA level of hypoxia-inducible factor 1α (HIF-1α) was detected by real-time quantitative PCR (qPCR). HIF-1α protein expression was detected by Western blotting. Anti-AS activity of effective compounds were evaluated. Results: Among the 200 compounds, 11 compounds could significantly inhibit the increase of luciferase activity in THP-1-HIF-1α-HER-Luciferase cells induced by hypoxia (all P<0.05), and compound numbered 14 (C14) had the most significant inhibitory effect. THP-1 and U937 cells formed foam cells induced for 24 h by OX-LDL. However, cells were pretreated with C14 for 2 h, which could significantly inhibit the formation of foam cells induced by OX-LDL. Cells were induced for 24 h by OX-LDL, which could significantly increase the expression of HIF-1α mRNA and protein (all P<0.05), while cells pretreated with C14 could significantly inhibit the increase of HIF-1α mRNA and protein expression in a gradient-dependent manner (all P<0.05). Conclusion: THP-1-HIF-1α-HER-Luciferase high-throughput model can be reliability used in screening of compounds with anti-AS activity. C14 has the good anti-AS activity characteristics.
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AIM To investigate the effect of Shenfu Injection on atherosclerosis (AS) models of high-fat apolipoprotein E-deficient (ApoE-/-) mice,and to explore its anti-atherosclerosis mechanism through the observation of oxidative stress (OS) variation.METHODS C57 mice were used as controls.ApoE-/-mice fed with 20-week high fat diet were randomly divided into model group,Shenfu group for subsequent 4-week continuous corresponding intervention,after which the mice had their blood lipid levels measured,their levels of MPO and NOX4 identified by ELISA,and their T-SOD activity determined by hydroxylamine method,their MDA level detected by TBA,their plaque formation observation achieved by HE staining of aortic gross and red O of all the aorta,and their Nrf2 mRNA expression detected by real time qPCR method.RESULTS Compared with the control group,the model group manifested with increased contents of TG,TC,LDL,decreased HLD;decreased activity of SOD,increased contents of MPO,NOX4 and MDA,and down-regulated expression of aortic Nrf2 and Keap1 mRNA.Compared with the model group,Shenfu Injection group was observed with no obvious blood lipid level change,but a reduction of plaque area,and an effective inhibition on OS as revealed by improved levels of Nrf2 and Keap1 mRNA.CONCLUSION Shenfu Injection can activate Nrf2 and interfer the relevant enzymes,thus prevents the atherosclerosis progression through OS reduction.
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Atherosclerosis is an inflammatory disease of arterial wall caused by many factors, which is the main pathological basis of many cardiovascular diseases. Currently, "inflammatory response" theory is widely accepted as pathogenesis of atherosclerosis. Abnormal increase of apolipoprotein ApoB-100, a composition of low density lipoprotein (LDL), which causes pathological inflammation, is a major factor leading to atherosclerosis. Therefore, inhibition of ApoB-100 induced pathological inflammatory response by immunotherapy is expected to delay the development of atherosclerosis. This review focused on the recent advances of ApoB-100 vaccine and other ApoB-100 inhibitors against atherosclerosis.
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Objective In the computational fluid dynamics software FLUENT, the independently developed user defined function (UDF) dynamic mesh program is called to achieve the mobile update of grid note based on the wall shear stress (WSS). Then this method is applied to simulate the development process of atherosclerosis (AS). Methods The UDF program by secondary development could extract WSS results of every note on the wall during the computing process, and if the threshold value criterion condition was met, the node would be adjusted to a new position. The mesh regeneration method combining with the spring smoothing and the local remeshing was adopted to control the update of the grid, so as to ensure the grid quality during deformation. Results The UDF program successfully extracted the WSS and arranged the corresponding deformation for the grid. The morphology of local extension in the proximal part and restenosis in the distal end were resulted from the vortex in the rear of the initial stenosis. Those features were similar to the indication of clinical angiography. Conclusions The independently developed UDF program has reached the expected effects, depicting the topography characteristics of AS influenced by WSS. In future researches, more influential factors should be considered in dynamic mesh deformation control to provide numerical references for clinical prognosis and risk evaluation of AS.
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Chronic intermittent hypoxia ( CIH ) caused by ob-structive sleep apnea hypopnea syndrome( OSAHS) is an impor-tant factor causing or aggravating many kinds of cardiovascular and cerebrovascular diseases. Establishing a rational animal model for intermittent hypoxia is an essential method to study the CIH related cardiovascular diseases. Recently, researchers have tended to simulate intermittent hypoxia condition by controlling the oxygen concentration of the environmental air around the ani-mals. In the paper, we summarize and compare the methods of making intermittent hypoxia animal model in recent literature, from aspects of experimental animals, gas control apparatus, gas species and concentration, intermittent hypoxia treatment time, and anoxic cycle mode.
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OBJECTIVE:To study the effects of Total sasponins of Pannax Notoginseng(PSN)on oxidative stress during atherosclerogenesis(AS)in rabbits accelerated by persistent inflammation.METHODS:A total of 24 Japanese big-eared rabbits were divided into control group,high lipid group,persistent inflammatory group,and PSN group to be treated accordingly.Serum levels of C reactive protein(CRP),nitric oxide(NO)and malondialdehyde(MDA)as well as the activity of superoxide dismutase(SOD)were given a dynamic monitoring;and aortic atherosclerotic plaque areas were measured.RESULTS:In the persistent inflammatory group,serum levels of CRP and MDA were increased significantly,but SOD activity and level of NO were decreased significantly,aortic atherosclerotic plaque area was increased obviously,and CRP level was positively correlated with both MDA content and plaque area,showing significant difference as compared with the high-lipid group(P
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Objective: To study the prevention and treatment of photosynthetic bacteria (PSB) on atherosclerosis (AS). Method: The hypolipidemic effect of PSB on atherosclerosis (AS) in rabbits induced by high cholesterol diet was studied with Spirulina platensis as positive control. The content of liver TC,AS plaque area percentage (PA) and the emphraxis of coronary artery were determined. Results:PSB could significantly reduce the content of TC in liver, the liver index (LW/BW), and PA 63.99%, 21.96%, 70.10%, respectively, and remarkably abate the lipid aggregation in liver and the pathological changes of coronary artery. In comparison,PSB was better than Spirulina platensis. Conclusion:PSB can prevent the formation of atherosclerosis in hyperlipidemic rabbits.
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Atherosclerosis (AS) is pathologically important basis of many kinds of coronary atherosclerosis disease (CAD). It can be substantially protected by raising high-density lipoprotein (HDL).In view of mechanism, drugs for raising HDL include: cholesterol ester transfer protein inhibitors, peroxisomal proliferator-activated receptor agonists, liver X-activated receptor agonists, farnesoid X receptor antagonists or agonists, lipoprotein lipase activators, niacin, and phenytoin and lecin : cholesterol acyltransferase activators, etc. This review aimed to the progress of drugs for regulating high-density lipoprotein and their mechanism, in view of clinical and preclinical aspects.
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Aim To study on the protective effects on vein endothelial cell of scallop skite-glycosaminoglycan(SS-GAG)and the mechanism of anti-atherosclerosis action of SS-GAG.Methods The endothelial cell of human umbilical vein had been cultured in vitro, and we established an model of endothelial cell oxidative damage induced by oxidized low density lipoprotein (OX-LDL), MTT assay and chemical methods were used to test the influence of SS-GAG on proliferation activity of endothelial cell oxidative damage and analyze nitric oxide (NO) and endothelial nitric oxide synthase (eNOS).Results Oxidized lowdensity lipoprotein (OX-LDL) remarkably inhibited the ability of cell proliferation, decreased nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) (P
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Objective To observe the effects of Chlamys farreri skirt glycosaminoglycan(SS-GAG) on the expressions of c-myc and TNF-? genes so as to explore the anti-atherosclerosis(AS) mechanism of SS-GAG.Methods The cell proliferation model of vascular smooth muscle cells(VSMC) was established by basic fibroblast growth factor(bFGF) induction.In Situ hybridization marked by non-isotope was applied to determine the effects of SS-GAG on the mRNA levels of c-myc and TNF in proliferative VSMC.Results The mRNA levels of c-myc and TNF in low dose group and high group of SS-GAG were obviously lower than that of model group(P
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As an important lipid in cell membrane and lipoprotein,sphingomyelin(SM)has widespread and considerable biological functions.It has close relation to the occurrence and development of atherosclerosis(AS),and the change of its concentration has become a seperate risk of atherosclerosis.Sphingomyelin synathase(SMS),the key enzyme in synthesis of SM,might become a new target for treatment of AS because of its expression and activity could directly influence the pathophysilogical process of AS.Here the recent reports about the structure and biological functions of SMS are summarized,and its role in AS and the possibilities of it being a potential drug target for AS therapy are reviewed.