Mechanism of Renshentang in Treatment of Atherosclerosis Based on Autophagic Effect of TRPV1 on Arterial Smooth Muscle / 中国实验方剂学杂志

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.

Compound Danshen Dripping Pill inhibits hypercholesterolemia/atherosclerosis-induced heart failure in ApoE and LDLR dual deficient mice via multiple mechanisms

Heart failure is the leading cause of death worldwide. Compound Danshen Dripping Pill (CDDP) or CDDP combined with simvastatin has been widely used to treat patients with myocardial infarction and other cardiovascular diseases in China. However, the effect of CDDP on hypercholesterolemia/atherosclerosis-induced heart failure is unknown. We constructed a new model of heart failure induced by hypercholesterolemia/atherosclerosis in apolipoprotein E (ApoE) and LDL receptor (LDLR) dual deficient (ApoE-/-LDLR-/-) mice and investigated the effect of CDDP or CDDP plus a low dose of simvastatin on the heart failure. CDDP or CDDP plus a low dose of simvastatin inhibited heart injury by multiple actions including anti-myocardial dysfunction and anti-fibrosis. Mechanistically, both Wnt and lysine-specific demethylase 4A (KDM4A) pathways were significantly activated in mice with heart injury. Conversely, CDDP or CDDP plus a low dose of simvastatin inhibited Wnt pathway by markedly up-regulating expression of Wnt inhibitors. While the anti-inflammation and anti-oxidative stress by CDDP were achieved by inhibiting KDM4A expression and activity. In addition, CDDP attenuated simvastatin-induced myolysis in skeletal muscle. Taken together, our study suggests that CDDP or CDDP plus a low dose of simvastatin can be an effective therapy to reduce hypercholesterolemia/atherosclerosis-induced heart failure.

Human 8-cell embryos enable efficient induction of disease-preventive mutations without off-target effect by cytosine base editor

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.

Association of Circulating Matrix Metalloproteinase-9 and the Risk of Coronary Heart Disease in Young Smokers

@#Introduction: Smoking causes cardiovascular risk which may alter the stability between the production and degradation of the extracellular matrix. Matrix metalloproteinase-9 (MMP-9) is a zinc-containing endopeptidase that degrades the extracellular matrix and plays a vital role in tissue remodeling. As a result, elevated serum MMP-9 levels produced by smoking, particularly at young age, raise the risk of future CHD. So this study aims to find out the possible relationship between circulating MMP-9 and the risk of cardiovascular disease in young smokers. Methods: The study was conducted on smokers with CHD subjects attending cardiology and medicine OP of the SRM Medical College Hospital and research center Tamil Nadu, India. The study group was divided into three groups. Group 1 includes 120 healthy controls as nonsmokers, Group 2 includes 120 smokers with Coronary heart disease (CHD), and Group 3 includes 120 smokers with diabetes and CHD subjects in the age group of 20-55 years. Serum MMP-9, hs-CRP, and APO-E levels were measured using the ELISA method and the lipid level was measured enzymatically using AU480 automatic analyzer (back man coulter). Results: The mean serum MMP-9, hs-CRP, and APO-E levels were significantly higher in both groups (p<0.05) when compared to controls. The study also shows a significant positive association between MMP-9 with hs-CRP, APO-E, smoking burden, and smoking intensity. Conclusion: The study concludes a significant association exists between cigarette smoking with MMP-9 and also relative exposure to circulating inflammation markers plays a potential role in the pathogenesis of CHD.

Polymorphisms of apolipoprotein E in the Afro-descendant population of Buenaventura, Colombia / Polimorfismos del gen de la apopoliproteína E en la población afrodescendiente de Buenaventura, Colombia

ABSTRACT Objetives: To estimate the frequency distribution, both allelic and genotypic, of the APOE gene in the Afro-descendant population of Buenaventura, Colombia. Methods: Three hundred and forty-eight Afro-descendant individuals were analyzed and the APOE locus was genotyped by PCR-RFLP. The allelic and genotypic frequencies were established by direct counting and the Hardy-Weinberg equilibrium was evaluated through X2 test. The frequencies obtained in this study were compared with frequencies reported for other Colombian populations through the Fisher's exact test. Results: The following allelic frequencies were observed: E3, 70.8%; E4, 21.4%, and E2, 7.8%. The genotypic frequencies were: E3/E3, 51.1%; E3/E4, 27.3%; E2/E3, 12.1%; E4/E4, 6%; E2/E4, 3.5%, and E2/E2, 0%. The entire examined population was found in Hardy-Weinberg equilibrium (P = .074), and significant differences were found in the allele E4 when comparing this population with the Amerindian and mestizo populations of Bogotá, Quindío, Centro-Oriente, Valle del Cauca, Barranquilla and Medellín (P< 0.0345). Conclusions: The allelic frequencies observed in this study were significantly different from the frequencies reported in other Colombian populations. The high representativeness of the E4 and E2 alleles validates the hypothesis that there are micro-evolutionary processes that have been acting on their frequencies and could be associated with susceptibility to neuropsychiatric diseases such as Alzheimer's disease, metabolic alterations of fats and/or coronary artery disease.

RESUMEN Objetivos: Estimar la distribución de frecuencias tanto alélicas como genotípicas del gen APOE en la población afrodescendiente de Buenaventura, Colombia. Métodos: Mediante la técnica de PCR-RFLP's se analizaron 348 individuos no relacionados de esta ciudad. Se realizó el cálculo de frecuencias alélicas y genotípicas y se evaluó el equilibrio de Hardy-Weinberg mediante la prueba de la X2. Se compararon las frecuencias alélicas obtenidas en el presente estudio con otras poblaciones de Colombia mediante el test exacto de Fisher. Resultados: Se reportaron las siguientes frecuencias alélicas: E2, 7,8%; E3, 70,8%, y E4, 21,4%. Las frecuencias genotípicas fueron: E3/E3, 51,1%; E3/E4,27,3%; E4/E4,6%; E2/E3,12,1%; E2/E4, 3,5%, y E2/E2, 0%. La población total se encontró en equilibrio de Hardy-Weinberg (p = 0,074), y se hallaron diferencias significativas en el alelo E4 al comparar esta población con las amerindias y mestizas de Bogotá, Quindío, Centro-Oriente, Valle del Cauca, Barranquilla y Medellín (p < 0,0345). Conclusiones: Las frecuencias alélicas observadas fueron significativamente diferentes de las frecuencias reportadas en otras poblaciones de Colombia. La alta representatividad de los alelos E4 y E2 validan la hipótesis de que hay procesos microevolutivos que han venido actuando en sus frecuencias y pueden estar asociadas con susceptibilidad a enfermedades neuropsiquiátricas como la enfermedad de Alzheimer, alteraciones metabólicas de las grasas y/o enfermedad coronaria.

Intervention of Huanglian Jiedutang on Atherosclerosis in ApoE-/- Mice and Its Mechanism / 中国实验方剂学杂志

ObjectiveTo study the intervention of Huanglian Jiedutang on atherosclerosis （AS） in apolipoprotein E knockout （ApoE-/-） mice induced by the high-fat diet. MethodThe ApoE-/- mouse model of AS was induced by the high-fat diet， and Huanglian Jiedutang was used to intervene in the AS in the ApoE-/- mice. The pathological changes of aorta were observed by hematoxylin-eosin （HE） staining. The levels of serum total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） were detected by an automatic biochemical analyzer. The protein expression levels of sirtuin-1 （SIRT1） and nuclear factor-kappa B （NF-κB） were determined by Western blot assay， and the mRNA expression levels of adenosine 5'-monophosphate-activated protein kinase （AMPK）， peroxisome proliferators-activated receptors α （PPARα）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and NOD-like receptor pyrin domain-containing 3 （NLRP3） were determined by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultAs compared with the normal group， there was a large amount of lipid accumulation in the blood vessels of the model group. In the model group， the levels of serum TG， TC， and LDL-C were increased （P<0.01）， and the level of HDL-C was decreased （P<0.01）. The protein expression level of SIRT1 in the aorta was decreased， while that of NF-κB was increased in the model group （P<0.01）. The mRNA expression levels of IL-6， TNF-α， and IL-1β were higher （P<0.01）， while those of AMPK in the liver were lower in the model group （P<0.01）. Compared with the model group， the Huanglian Jiedutang group reduced the lipid accumulation and inflammatory reaction in the aorta of mice with AS， reduced the levels of TC， TG， and LDL-C （P<0.01）， and increased the level of HDL-C （P<0.01）. Huanglian Jiedutang significantly increased the protein expression level of SIRT1 in the aorta of ApoE-/- mice （P<0.01） and decreased the protein expression levels of NF-κB in the aorta （P<0.05， P<0.01）. Huanglian Jiedutang down-regulated the mRNA expression levels of TNF-α， IL-6， IL-1β， and NLRP3 in the aorta （P<0.05， P<0.01）， and up-regulated the mRNA expression levels of AMPK and PPARα in the liver of ApoE-/- mice （P<0.05， P<0.01）. ConclusionHuanglian Jiedutang has a certain intervention effect on the formation of atherosclerotic aortic plaque in ApoE-/- mice. Its mechanism may be related to the decrease of serum TC， TG， and LDL-C levels， the increase of HDL-C levels， thus playing a role in lowering blood lipid， the increase of SIRT1 protein， the decrease of NF-κB protein， the decrease of inflammatory factors such as TNF-α and IL-6， which protects blood vessels from inflammatory injury， and the improvement of AMPK and PPARα levels to participate in autophagy and apoptosis.

Effect of Ethyl Acetate Extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on Intestinal Flora in ApoE-/- Mice with Atherosclerosis / 中国实验方剂学杂志

ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout （ApoE-/-） mice， and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group， rosuvastatin group （10 mg·kg-1）， high-， low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis （75， 25 mg·kg-1）， with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration， blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin （HE） staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group， the plaque area of the model group was significantly increased with inflammatory infiltration， the contents of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， inflammatory factors and inducible nitric oxide synthase （iNOS） were increased， while the content of high-density lipoprotein cholesterol （HDL-C） was decreased. Compared with the model group， rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque， reduce the contents of TG， TC， LDL-C， inflammatory factors and iNOS， and increase the content of HDL-C. Compared with the blank group， the relative abundances of Firmicutes and Proteobacteria in the model group increased， while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated， and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group， ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids， which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors， improve antioxidant capacity and repair the disorder of intestinal flora structure.

Effect and Mechanism of Longshengzhi Capsule on Atherosclerosis in ApoE-/- Mice / 中国实验方剂学杂志

ObjectiveTo study the effect of Longshengzhi capsule （LSZC） on high fat diet （HFD）-induced atherosclerosis （AS） in apolipoprotein E knockout （ApoE-/-） mice. MethodApoE-/- mice were fed with HFD for 8 weeks to induce AS. Then the mice were randomized into model group， simvastatin group （4 mg·kg-1）， high-dose LSZC group （1.6 g·kg-1）， medium-dose LSZC group （0.8 g·kg-1）， and low-dose LSZC group （0.4 g·kg-1）. C57BL/6J Mice with normal diet were used as the blank control. After 10 weeks， serum levels of triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， malondialdehyde （MDA）， superoxide dismutase （SOD）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） were detected. Hematoxylin-eosin （HE） and oil red O were used to detect aortic plaque in each group. The levels of CD34 and F4/80 in aorta were determined by immunohistochemistry （IHC）. ResultCompared with the blank control， the model group demonstrated obvious aortic plaque， a large amount of lipid accumulation， serious damage of aortic intima， increase in serum levels of TC， TG， LDL-C， HDL-C， MDA， IL-1β， and IL-6 （P<0.01）， decrease in SOD level （P<0.01）， and rise of the expression of CD34 and F4/80 （P<0.01）. Compared with the model group， LSZC of the three doses all decreased the serum levels of TG and LDL-C （P<0.05）， and the levels of IL-1β and IL-6 （P<0.05， P<0.01）， and the high-dose and medium-dose LSZC improved SOD level， decreased MDA content （P<0.05， P<0.01）， and reduced the expression of the CD34 and F4/80 in blood vessels （P<0.05， P<0.01）. ConclusionLSZC has certain intervention effect on the formation of aortic plaque in atherosclerosis ApoE-/- mice. The mechanism is that it reduces the levels of serum TG and LDL-C to lower blood lipid， decreases MDA level and improves SOD activity to inhibit lipid peroxidation， lowers the levels of IL-1β and IL-6 and down-regulates the expression of CD34 and F4/80 to protect blood vessels from inflammatory damage.

Preliminary study on the polymorphisms of some related genes and the methylation levels of BAX and ApoE genes in Alzheimer′s disease / 中华神经科杂志

Objective:To preliminarily explore the association between single nucleotide polymorphisms (SNP) of five candidate genes (APH1B, PRNP, HMGCR, SIRT1, ApoE) and Alzheimer′s disease (AD), and to analyze the methylation levels of BAX and ApoE promoters on the pathogenesis of AD.Methods:Seventeen cases who were admitted to the Department of Geriatrics of the First Affiliated Hospital of Xinjiang Medical University from 2014 to 2015 and diagnosed as likely to be AD by geriatrician and neurologists according to the AD diagnostic criteria in 4th Revised Edition of the Diagnostic and Statistical Manual of Mental Disorders of the American Psychiatric Association served AD group, with an age of (75.65±5.86) years, and 34 non-AD patients with matching baseline data such as age, gender, ethnicity, and education status among patients hospitalized during the same period were selected as control group, with an age of (77.59±7.41) years. Sanger sequencing method was used for SNP typing of candidate genes. Methylation-specific polymerase chain reaction was used to determine the DNA methylation level.Results:The distribution of ApoE ε4 allele was statistically different between the AD group and the control group (χ 2=9.718, P=0.002). Candidate genes (SIRT1 rs7895833, APH1B rs1047552, PRNP rs1799990, HMGCR rs3846662) SNP locus genotypes and alleles had no statistically significant differences in the distribution between the AD group and the control group ( P>0.05). After stratification according to whether they carried ApoE ε4, no statistically significant difference was found between the two groups ( P>0.05). The BAX promoter methylation level of the AD group (0.045±0.025) was lower than that of the control group (0.061±0.028) ( t=-2.078, P=0.045). After gender stratification, the BAX methylation level of the female AD group (0.044±0.021) was lower than that of the control group (0.065±0.275) ( t=-2.230, P=0.045). There was no statistically significant difference in the methylation level of ApoE promoter between the AD group and the control group ( P>0.05). After stratification according to whether they carry ApoE ε4 or not, the methylation level of AD patients with ApoE ε4 allele (1.553±0.291) was higher than that of non-carriers (1.221±0.261) ( t=2.480, P=0.025). Conclusions:ApoE ε4 allele may be a risk factor for the onset of AD. BAX promoter hypomethylation contributes to AD in the elderly in Xinjiang, especially in female. ApoE ε4 allele may cause AD through the interaction with ApoE methylation.

Effect and Mechanism of Xiangsha Liujunzi Tang on Lipid Deposition in Liver of ApoE-/- AS Mice by Affecting Long Noncoding RNA-HC/miR-130b and to Regulate Cholesterol Metabolism / 中国实验方剂学杂志

Objective:To investigate the mechanism of Xiangsha Liujunzi Tang in improving liver lipid deposition in ApoE^-/- atherosclerotic （AS） mice by affecting long noncoding RNA-HC （Lnc-HC）/microRNA-130b （miR-130b） in the regulation of cholesterol metabolism. Method:Totolly 10 C57BL/6J mice were selected as normal controls， and 30 healthy ApoE^-/- mice fed with high fat diet for 12 weeks were then randomly divided into the model group， Xiangsha Liujunzi Tang group（19.12 g·kg^-1·d^-1） and simvastatin group（2.275 mg·kg^-1·d^-1）， with gavage administration for 4 weeks. The serum lipid level of mice was detected by automatic biochemistry analyzer， and the histopathological changes of liver cells were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect expression of long noncoding RNA-HC， and miR-130b. Real-time PCR and Western blot assay were used to detect gene and protein expression of peroxisome proliferator-activated receptor gamma （PPAR<italic>γ</italic>）， liver X receptor （LXR）， ATP-binding cassette transporters A1 （ABCA1）， ATP-binding cassette transporters G1 （ABCG1）， ATP-binding cassette transporters G5 （ABCG5）， and ATP-binding cassette transporters G8 （ABCG8）. Result:Compared with the normal control group， the mice in the model group showed abnormal blood lipids， larger liver cells， obvious fat vacuoles， significantly increased expression of Lnc-HC， miR-130b in liver， and significantly decreased gene and protein expression of PPAR<italic>γ</italic>， LXR， ABCA1， ABCG1， ABCG5， and ABCG8 in mice liver （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with the model group， the abnormal blood lipid levels of the mice in the Xiangsha Liujunzi Tang group and the simvastatin group were improved， and the number of fatty vacuoles of liver cells was significantly reduced， the expression of liver Lnc-HC， miR-130b in Xiangsha Liujunzi Tang group decreased significantly （<italic>P</italic><0.05，<italic>P</italic><0.01）， the gene and protein levels of liver PPAR<italic>γ</italic>， ABCA1， ABCG1， ABCG5， ABCG8 in mice of the Xiangsha Liujunzi Tang group and the simvastatin group showed an upward trend. Among them， the gene and protein expression of LXR protein in the liver of the Xiangsha Liujunzi Tang group was significantly up-regulated （<italic>P</italic><0.05）. Conclusion:Xiangsha Liujunzi Tang may improve the lipid deposition in the liver of ApoE^-/-AS mice by affecting Lnc-HC/miR-130b to regulate the cholesterol metabolism process mediated by PPAR<italic>γ</italic>， thus playing a role in preventing and treating AS.

Effect and Mechanism of Banxia Baizhu Tianmatang on Atherosclerosis in ApoE-/- Mice / 中国实验方剂学杂志

Objective:To study the effects of Banxia Baizhu Tianmatang （BBTT） on atherosclerosis in apolipoprotein-E knockout （ApoE^-/-） mice induced by high fat diet. Method:The atherosclerosis model of ApoE^-/- mice was established with high-fat diet， and BBTT was used for intervention. The pathological changes of aorta after atherosclerosis were observed by hematoxylin-eosin （HE）， oil red O and Masson staining. The changes of serum total cholesterol （TC）， triglyceride （TG）， high density lipoprotein cholesterol （HDL-C） and low density lipoprotein cholesterol （LDL-C） were detected by automatic biochemical analyzer. The expression levels of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-6 （IL-6） and oxidized low density lipoprotein （ox-LDL） were detected by enzyme linked immunosorbent assay （ELISA）. Total tissue proteins were extracted， quantified by protein quantification （BCA） method， and the expression of matrix metalloproteinase-9 （MMP-9） protein was detected by Western blot. Thiobarbituric acid （TBA） method was used to detect the change of malondialdehyde （MDA） content. The change of superoxide dismutase （SOD） activity was detected by 2-（2-methoxy-4-nitrophenyl）-3-（4-nitrophenyl）-5-（2，4-disulfobenzene）-2H tetrazole monosodium salt （WST-8） method. Result:Compared with the control group， there was a large amount of lipid accumulation in the blood vessels of the model group， the serum levels of TG， TC， HDL-C and LDL-C significantly increased （<italic>P</italic><0.05）， the expression of MMP-9 protein in the blood vessels significantly increased （<italic>P</italic><0.01）， the expression levels of IL-6 and TNF-<italic>α</italic> in the serum increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， the SOD activity was significantly reduced （<italic>P</italic><0.01）， and the levels of MDA and ox-LDL expression increased （<italic>P</italic><0.01）. Compared with the model group， the treatment with BBTT could inhibit the accumulation of lipids in blood vessels， the TG levels were reduced in the high and medium dose groups of BBTT （<italic>P</italic><0.05）， high， medium and low dose groups significantly reduced the levels of LDL-C in serum （<italic>P</italic><0.01）， the expression of MMP-9 protein in blood vessels （<italic>P</italic><0.05） and IL-6 in serum （<italic>P</italic><0.01）， the high-dose group down-regulated the expression of TNF-<italic>α</italic> in serum （<italic>P</italic><0.01） and ox-LDL （<italic>P</italic><0.01）， both the high and medium-dose groups increased the level of MDA （<italic>P</italic><0.05， <italic>P</italic><0.01） and the activity of SOD （<italic>P</italic><0.05）. Conclusion:BBTT has a certain intervention effect on the formation of atherosclerosis aortic plaque in ApoE^-/- mice， and its mechanism may be associated with reducing the TG and LDL-C levels， lowering blood lipid， down-regulating MMP-9 protein， protecting blood vessels from inflammatory damage， reducing ox-LDL and MDA levels， and improving SOD activity to play an antioxidant role.

Effects of Total Flavonoids from Chamomile on Lipid Metabolism of Hyperlipidemia Model Mice and Its Mechanism / 中国药房

OBJECTIVE：To st udy t he effects of total flavonoids from chamomile on lipid metabolism of hyperlipidemia model mice and its potential mechanism. METHODS ：Thirty male C 57BL/6J-ApoE－/－ mice were randomly divided into model group ， positive control group（fenofibrate 30 mg/kg）and chamomile total flavonoids low-dose ，medium-dose and high-dose groups （88， 176，352 mg/kg），with 6 mice in each group. In addition ，6 male C 57BL/6J mice were used as normal control group. Mice in normal control group were fed with ordinary diet ，and mice in other groups were fed with high-fat diet for 8 weeks to replicate hyperlipidemia model. At the time of making model ，administration groups were given relevant liquid （using 1％ sodium carboxymethyl cellulose as solvent ）；normal control group and model group were given 1％ sodium carboxymethyl cellulose intragastrically，200 mL per gavage ，once a day ，for consecutive 8 weeks. The body weight of mice in each group was weighed before medication and 8 weeks after medication. The serum contents of total cholesterol （TC），triacylglycerol（TG），low-density lipoprotein cholesterol （LDL-C），high-density lipoprotein cholesterol （HDL-C），aspartate aminotransferase （AST）and alanine aminotransferase （ALT） in mice were detected after last administration ；the contents of superoxide dismutase （SOD） and malondialdehyde（MDA）as well as the protein expressions of peroxisome proliferator-activated receptor α（PPARα），carnitine palmityl transferase 1A（CPT1A）and peroxase acyl-CoA oxidase 1（ACOX1）in liver tissue were determined. The pathological changes i n liver tissue were observed. RESULTS：Compared w ith before medication ，the body weight of each group showed an increasing trend after 8 weeks of medication. Compared with normal control group ，body weight ，the contents of TC ，TG， LDL-C，AST and ALT in serum and MDA content in live r lan- tissue of mice in model group were significantly increased wei516@sina.com after 8 weeks of medication （P＜0.05 or P＜0.01）. The ·2706· China Pharmacy content of HDL-C in serum and the cont ent of SOD in liver tissue ，as well as the protein expressions of PPARα，CPT1A and ACOX1 were significantly decreased （P＜0.05 or P＜0.01），and the structure of liver tissue was disorganized ，with circular fat vacuoles of different sizes and lipid droplets of different sizes in the cytoplasm. Compared with model group ，body weight （except for chamomile total flavonoids low-dose group ）of mice ，serum contents of TC ，TG，LDL-C，AST and ALT ，content of MDA in liver tissue （except for chamomile total flavonoids low-dose and medium-dose groups ）were significantly decreased （P＜0.05 or P＜ 0.01）. Serum content of HDL-C ，content of SOD in liver tissue ，protein expressions of PPARα，CPT1A（except for chamomile total flavonoids low-dose and medium-dose groups ） and ACOX 1 were significantly increased （P＜0.05 or P＜0.01）；liver tissue structure was clear ，and liver fat vacuoles were improved to varying degrees ，and less lipid droplets. The improvement effect of the above indexes was the best in the chamomile total flavonoids high-dose group. CONCLUSIONS ：Chamomile total flavonoids can prevent the occurrence of hyperlipidemia in C57BL/6J-ApoE －/－ mice，the mechanism of which may be associated with up-regulation of PPARα expression，the improvement of liver injury and oxidant stress injury.

Recent advances in nanomedicines for the treatment of ischemic stroke

Ischemic stroke is a cerebrovascular disease normally caused by interrupted blood supply to the brain. Ischemia would initiate the cascade reaction consisted of multiple biochemical events in the damaged areas of the brain, where the ischemic cascade eventually leads to cell death and brain infarction. Extensive researches focusing on different stages of the cascade reaction have been conducted with the aim of curing ischemic stroke. However, traditional treatment methods based on antithrombotic therapy and neuroprotective therapy are greatly limited for their poor safety and treatment efficacy. Nanomedicine provides new possibilities for treating stroke as they could improve the pharmacokinetic behavior of drugs

Effects of SLCO1B1 521 T>C and APOE gene polymorphisms on lipid-lowering efficacy and adverse reactions of atorvastatin / 药学实践杂志

Objective To study the effect of SLCO1B1 521 T>C and APOE gene polymorphisms on the clinical efficacy and safety of atorvastatin in ischemic stroke patients with dyslipidemia. Methods 210 cases of ischemic stroke with dyslipidemia were enrolled from April 2018 to December 2018 to determine SLCO1B1 521 T>C and APOE gene polymorphisms. Patients received atorvastatin 20 mg/d orally. TC, TG, HDL-C, LDL-C levels were measured to evaluate the efficacy 3 months pre-and post- treatment. TBil, ALT, AST, CK levels were assayed with following up adverse reactions to evaluate safety. Results SLCO1B1 521 T>C genotype distribution was TT79.05%, TC19.05%, CC1.90%. E2, E3, E4 allele frequencies of APOE genes were 14.28%, 67.62%, 18.10%. Each genotype conforms to the law of Hardy-Weinberg balance. After three months of medication, there were significant differences in TC, TG, LDL-C, HDL-C changes in patients with different APOE genotypes. No obvious abnormality was found in safety index. The incidence of myalgia in SLCO1B1521 T>C mutant group was significantly higher than that in the wild group (P<0.01). Conclusion Lipid regulation of atorvastatin was affected by APOE gene polymorphism. SLCO1B1521 T>C may be associated with myalgia, the adverse reaction of atorvastatin. The detection of SLCO1B1 and APOE genotyping is helpful for individualized treatment of blood lipids and provides basis for rational use of statins in patients for drug therapy management.

Study on effect of "Trichosanthis Fructus-Allii Macrostemonis Bulbus" on atherosclerosis in ApoE~(-/-) mice based on liver metabonomics / 中国中药杂志

In this study, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS)-based liver metabolomics approach was used to explore the mechanism of "Trichosanthis Fructus-Allii Macrostemonis Bulbus" in improving atherosclerosis(AS) of mice with apolipoprotein E gene knockout(ApoE~(-/-)). AS mouse model was induced by high-fat diet. The pathological and biochemical indexes such as the histopathological changes, body weight, liver weight, blood lipid level and inflammatory factors in the liver of mice were determined. The metabolic profiling of mice liver samples was performed with UPLC-Q-TOF-MS. Multiple statistical analysis methods including partial least squares discriminant analysis(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were employed to screen and identify biomarkers. The levels of related enzymes including LCAT, sPLA2, EPT1 and ACER1 were detected. The results showed that "Trichosanthis Fructus-Allii Macrostemonis Bulbus" significantly reduced the areas of aortic plaque and fat vacuoles of liver in AS mice and decreased the accumulation of lipid droplets and liver coefficient. "Trichosanthis Fructus-Allii Macrostemonis Bulbus" also regulated the levels of blood lipid and inflammatory injury in the liver. The metabolites of the control group, the model group and the "Trichosanthis Fructus-Allii Macrostemonis Bulbus" group could be distinguished significantly. Fifteen potential biomarkers related to AS were discovered and preliminarily identified, seven of which could be regulated by "Trichosanthis Fructus-Allii Macrostemonis Bulbus" in a trend of returning to normal. Metabolic pathway analysis screened out two major metabolic pathways. "Trichosanthis Fructus-Allii Macrostemonis Bulbus" obviously regulated the levels of LCAT, sPLA2, EPT1 and ACER1. It was inferred that "Trichosanthis Fructus-Allii Macrostemonis Bulbus" could play a major role in AS treatment by regulating glycerophospholipid and sphingolipid metabolism disorders in the liver, with the mechanism probably relating to the intervention of the expression of LCAT, sPLA2, EPT1 and ACER1.

Schisandrin C alleviates atherosclerosis and inflammation in ApoE-/-mice fed with arterial high-fat diet / 中国药理学通报

Aim To investigate the effects of schisandrin C on arterial inflammation and atherosclerosis of ApoE-/-mice fed with high-fat diet. Methods ApoE-/-mice were divided into high fat diet group (HFD) and high fat diet group + schisandrin group (HFD + Sch C). High fat diet and Sch C were given at the same time. After 12 weeks, mice were executed and arterial tissues were collected. RT-q PCR and Western blot were respectively used to detect the expressions of IL-6, TNF-α, ICAM-1, β-actin and the phosphorylation and expression of IκB-α in arteries. Oil red O staining and biochemical kit were respectively used to detect the lipid changes in aortic root and the blood lipid levels. Results Compared with HFD group, the area of atherosclerotic plaque in HFD + Sch C was reduced (P < 0. 05), but Sch C did not affect blood lipid levels. Compared with HFD group, the mRNA expression of TNF-α, IL-6 and ICAM-1 and the phosphorylation of IκB-α of arterial tissue in HFD + Sch Cgroup decreased (P < 0. 05). Conclusions Sch C could effectively alleviate atherosclerosis induced by high-fat diet in ApoE-/-mice, accompanied by alleviation of arterial inflammation.

Based on "intestinal flora-inflammation" pathway to investigate effect and mechanism of Danlou Tablet on atherosclerosis of ApoE-/- mice / 中草药

Objective: To investigate the efficacy and mechanism of Danlou Tablet against atherosclerosis model of ApoE-/- mice fed with high fat diet. Methods: C57BL/6J mice were used as controls and ApoE-/- mice were randomly divided into two groups after 24 weeks of high fat feeding, including the model group received saline and the treatment group received Danlou Tablet. Animals were executed after 8 weeks of treatment and serum was collected to measure blood lipids; Plaque formation in the aorta was observed by red O and HE staining; 16 S rRNA sequencing was used to analyze changes in intestinal flora, and GC-MS test for detection fecal SCFAs content, ELISA for the determination of serum LPS, and real time PCR for detection of mRNA expression. Results: Compared with the control group, the blood lipid levels were increased; intestinal flora was imbalance with increased harmful bacteria and reduced beneficial bacteria; The level of serum LPS and inflammation around the aorta were increased in the model group. Compared with the model group, the contents of TG, TC, LDL-C and the plaque area of Danlou Tablet group were decreased (P < 0.01); Danlou Tablet group can regulate intestinal flora, thus effectively reducing serum LPS and inflammatory factors TNF-α, ICAM-1 and IL-1β levels around the aorta (P < 0.01). Conclusion: Danlou Tablet exerts an anti-atherosclerosis action with favorable efficacy through restructing the intestinal flora stucture, inhibiting endotoxin releasing and constraining the inflammatory response induced by dysbacteriosis.

Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet / 中国药科大学学报

@#Atherosclerosis(AS), characterized with the accumulation of lipids on the vessel wall, is an immune-related inflammatory disease which promotes the progression of cardiovascular diseases(CVD). The imbalance of Treg/Th17 accelerates the progression of AS. Yangyin Huoxue Prescription(YHF)is an efficient traditional Chinese medicine used in the treatment of AS, but the effects of YHF on the balance of immunity have still not been clarified. This project was designed to investigate the effects of YHF on the imbalance of Treg/Th17 and AS in ApoE-/- mice induced by high-fat diet(HFD). ApoE-/- mice were given HFD to induce AS and administered low-dose YHF(18 g/kg)or high-dose YHF(36 g/kg)for 20 weeks. Atherosclerotic plaque area was analyzed by oil red O staining. Serum lipids were measured by biochemical kits. Treg or Th17 cells in peripheral blood were detected by flow cytometry. mRNA and protein expression of Foxp3 and RORγt of aortas were determined by qRT-PCR, Western blot and immunohistochemistry. Splenic CD4+T cells of mice were isolated and activated by anti-CD3/CD28, and then treated with lipopolysaccharide(LPS)and YHF. The expression of mRNA and protein of Foxp3 and RORγt were detected by qRT-PCR and immunofluorescence. It was found that YHF reduced the plaque area, decreased lipid level and increased the ratio of Treg cells in peripheral blood. Moreover, YHF increased mRNA or protein expression of Foxp3 in aortas in vivo or CD4+T cells in vitro while decreasing mRNA or protein expression of RORγt. These results suggested that YHF can regulate the imbalance of Treg/Th17 in ApoE-/- mice induced by HFD, and reduce the inflammatory stimulation of LPS on CD4+T cells, thereby improving AS.

Metformin mediated reverse cholesterol transport to inhibit atherosclerosis / 中国药理学通报

Aim To investigate the molecular mechanism of metformin inhibiting atherosclerosis in ApoE 1_ mice by reverse cholesterol transport. Methods Eighteen ApoE -/ _ mice were randomly divided into three groups, control group, model group and metformin group, and body weight changes were monitored weekly. Blood samples were taken to measure serum lipid levels; animal ultrasound was used to measure abdominal aortic wall thickness; HE and oil red 0 staining were used to evaluate the degree of liver steatosis; Western blot was used to detect the expression of liver cholesterol reverse transport-related proteins LXRa and ABCA1. Results Compared with control group, the body weight, serum T C, T G, and LDL in model group increased, HDL decreased(P <0. 05), abdominal aortic wall thickened (P <0. 05), liver fat deposition increased, and L X R a, ABCA1 expression was reduced. In metformin group, body weight, serum T C, T G, LDL decreased, HDL increased (P <0. 05), liver fat deposition and abdominal aortic wall thickness were significandy reduced (P <0. 05), and LXRa and ABCA1 expressions markedly increased (P <0. 0 5). Conclusions Metformin can delay the progression of atherosclerosis by up-regulating the expression of liver cholesterol reverse transport related proteins LXRa and ABCA1, enhancing liver reverse cholesterol transport, regulating blood lipid metabolism and reducing liver lipid deposition.

Effect of aldehyde scavenger-salicylamine on atherosclerosis of uremic ApoE-/- mice / 西安交通大学学报(医学版)

Objective: To observe the effect of aldehyde scavenger, salicylamine (SAM), on atherosclerosis (AS) and its phenotype in uremic apolipoprotein E knockout (ApoE-/-) mice. Methods:Uremic ApoE-/- mice model was created by 5/6 nephrectomy; control ApoE-/- mice were sham-operated. Three subgroups of experimental mice were set up: uremia SAM intervention group, uremia group and control group, which were treated with SAM (1 g/L) or vehicle for 6 weeks, respectively. After the intervention was completed, we assessed the body weight, blood pressure, renal function, serum lipid profile, serum SAM concentration, extent and characteristic of aortic atherosclerotic lesion in each group of mice. Results: Compared with control group: aortic AS lesion area, necrotic area and macrophage content in AS lesion increased but collagen content in AS lesion decreased in uremia group. SAM treatment for 6 weeks lessened the atherosclerotic lesion area, necrotic area and macrophage content of plaques, and meanwhile increased collagen content of plaques in uremic mice, not accompanied by changes in body weight, blood pressure, serum lipid profile or renal function. SAM did not accumulate or induce toxic effect on uremic ApoE-/- mice. Conclusion: Aldehyde scavenger SAM ameliorates renal injury-induced acceleration of AS, alters atherosclerotic phenotype, and increases the stability of plaques. These benefits are independent of effects on blood pressure, lipid profile or renal function. SAM does not accumulate or induce toxic effect in uremic ApoE-/- mice.