Ethnopharmacological relevance Obesity-associated adipose tissue inflammation exacerbates atherosclerosis (AS) by inducing endothelial dysfunction. Simiaoyong'an Decoction (SMYA), a traditional Chinese medicine (TCM) is highly effective in managing AS-related cardiovascular diseases, but the underlying mechanisms are unclear. Purpose This study used network pharmacology, molecular docking, and AS model mice and cells to determine the mechanisms by which SMYA alleviates AS progression and attenuates visceral adipose tissue (VAT) inflammation. Methods Bioactive components of SMYA were identified using UHPLC-LTQ-Orbitrap-MS. An AS mouse model was established through high-fat-diet (HFD) feeding combined with perivascular carotid collar placement (PCCP). Therapeutic effects of SMYA were evaluated by histochemical staining, MOMA-2/α-SMA immunohistochemistry, and Enzyme-linked immunosorbent assay. SMYA targets against AS and adipose tissue inflammation were identified by network pharmacology and molecular docking, followed by mechanistic validation in animal models and an in vitro LPS-induced 3T3-L1-HUVECs coculture system. Western blotting analysis was used to elucidate the molecular mechanism underlying therapeutic effects of SMYA. Results UHPLC-LTQ-Orbitrap-MS analysis identified 23 bioactive components in SMYA. SMYA treatment significantly reduced plaque vulnerability and systemic inflammation (TNF-α and IL-6) in the AS model mice and cells. SMYA protected against endothelial cell injury by significantly enhancing PPARγ expression, increasing adiponectin secretion, inhibiting NF-κB, and downregulating endothelial injury markers (VCAM-1 and ICAM-1). These effects were reversed by PPARγ inhibitors. Conclusions SMYA protects vascular endothelial function and stabilizes atherosclerotic plaques by suppressing adipose tissue inflammation through PPARγ activation, thereby increasing adiponectin release. Therefore, SMYA is a promising new candidate for treating AS.

Background:Research has underscored the significance of targeting energy metabolic remodeling in heart failure (HF) as a crucial therapeutic avenue in recent years. Following the onset of heart failure, dysregulated energy metabolism induces multiple adverse effects, exemplified by the reduced expression of connexin 43 (Cx43)-a gap junction protein requiring substantial ATP for phosphorylation modification-in rats with post-myocardial infarction (MI) heart failure. In this study, we report that Wenxin Keli (also known as Wenxin granule), a clinically available Chinese patent medicine used for preventing and treating heart failure-related arrhythmias, modulates energy metabolism and improves Cx43 function by activating AMPK/SIRT1/PGC-1α signaling pathway. However, the pathological alterations after heart failure are intricate, and the underlying mechanism through which Wenxin Keli exerts its therapeutic effect on heart failure remains to be further elucidated. Methods:A post-myocardial infarction heart failure rat model was established via left anterior descending coronary artery ligation. Cardiac function was evaluated 4 weeks later using echocardiography, HE, and Masson trichrome staining. ELISA was employed to detect energy metabolism-related indices, while WB analysis was used to quantify the expression levels of proteins, including SIRT1, PGC-1α, and Cx43. IHC was further utilized to assess Cx43 protein content in tissue sections. Ventricular fibrillation (VF) was induced to determine the VF threshold, providing insights into arrhythmogenic susceptibility. Results:Wenxin Keli enhances energy metabolism and improves Cx43 function in post-MI heart failure rats by activating the AMPK/SIRT1/PGC-1α signaling pathway. Specifically, Wenxin Keli stimulates the SIRT1/PGC-1α axis, promoting interaction between PGC-1α and PPARs and ERRs. This dual mechanism addresses the combined impairments in fatty acid oxidation and glucose utilization after heart failure, restoring mitochondrial oxidative phosphorylation and increasing ATP production through the TCA cycle. Furthermore, Wenxin Keli boosts the positive regulatory effect of SIRT1 on PGC-1α by upregulating AMPK phosphorylation, thereby further activating the AMPK/SIRT1/PGC-1α signaling pathway and creating a positive feedback loop. Conclusion:Wenxin Keli exhibits multi-target regulation of energy metabolic disorders in post-myocardial infarction heart failure while protecting Cx43. Its core mechanism is activating the AMPK/SIRT1/PGC-1α signalling pathway and its downstream regulatory network.

To investigate the pharmacological mechanism of Qili Qiangxin Capsule (QLQX) improvement of heart failure (HF) based on miR133a-endoplasmic reticulum stress (ERS) pathway. A left coronary artery ligation-induced HF after myocardial infarction model was used in this study. Rats were randomly assigned to the sham group, the model group, the QLQX group [0.32 g/(kg·d)], and the captopril group [2.25 mg/(kg·d)], 15 rats per group, followed by 4 weeks of medication. Cardiac function such as left ventricular ejection fraction (EF), fractional shortening (FS), left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), the maximal rate of increase of left ventricular pressure (+dp/dt max), and the maximal rate of decrease of left ventricular pressure (−dp/dt max) were monitored by echocardiography and hemodynamics. Hematoxylin and eosin (HE) and Masson stainings were used to visualize pathological changes in myocardial tissue. The mRNA expression of miR133a, glucose-regulated protein78 (GRP78), inositol-requiring enzyme 1 (IRE1), activating transcription factor 6 (ATF6), X-box binding protein1 (XBP1), C/EBP homologous protein (CHOP) and Caspase 12 were detected by RT-PCR. The protein expression of GRP78, p-IRE1/IRE1 ratio, cleaved-ATF6, XBP1-s (the spliced form of XBP1), CHOP and Caspase 12 were detected by Western blot. TdT-mediated dUTP nick-end labeling (TUNEL) staining was used to detect the rate of apoptosis. QLQX significantly improved cardiac function as evidenced by increased EF, FS, LVSP, +dp/dt max, −dp/dt max, and decreased LVEDP (P<0.05, P<0.01). HE staining showed that QLQX ameliorated cardiac pathologic damage to some extent. Masson staining indicated that QLQX significantly reduced collagen volume fraction in myocardial tissue (P<0.01). Results from RT-PCR and Western blot showed that QLQX significantly increased the expression of miR133a and inhibited the mRNA expressions of GRP78, IRE1, ATF6 and XBP1, as well as decreased the protein expressions of GRP78, cleaved-ATF6 and XBP1-s and decreased p-IRE1/IRE1 ratio (P<0.05, P<0.01). Further studies showed that QLQX significantly reduced the expression of CHOP and Caspase12, resulting in a significant reduction in apoptosis rate (P<0.05, P<0.01). The pharmacological mechanism of QLQX in improving HF is partly attributed to its regulatory effect on the miR133a-IRE1/XBP1 pathway.

Objective: Erchen decoction, a traditional Chinese medicine formula, can reduce the level of oxidative stress for the treatment of dyslipidemia phlegm-dampness retention syndrome (DPDRS); however, studies have not elucidated the mechanism underlying its metabolic action. Here, liquid chromatography-mass spectrometry (LC-MS)-based metabolomic techniques were utilized to characterize the in vivo effects of Erchen decoction in achieving reduction of oxidative stress levels and understand the potential metabolic mechanisms of action. Methods: We constructed a DPDRS animal model using a multifactorial composite modeling approach, and Erchen decoction was administered by gavage. We employed LC-MS-based metabolomic techniques in combination with serum-associated factors, gene transcription, methylation detection, and hematoxylin and eosin staining. Results: In this study, the constructed animal model of DPDRS had satisfactory quality. Erchen decoction treat-ment reduced the levels of low-density lipoprotein cholesterol, t total cholesterol and riglyceride; it improved the endothelial structure, increased levels of serum beta-nicotinamide adenine dinucleotide phosphate and glutathione concentrations, increased aortic phosphoserine aminotransferase and phosphoserine phosphatase gene expression levels, and decreased aortic phosphoglycerate dehydrogenase methylation level. A total of 64 differential metabolites were obtained using LC-MS assay, and 34 differential metabolic pathways were obtained after enrichment. Conclusions: Erchen decoction treatment of DPDRS mice reversed lipid indexes, improved vascular endothelial structure, increased serum and aortic anti-oxidative stress factor concentration and expression levels, and decreased methylation levels, thereby reducing oxidative stress and protecting vascular endothelium. Tricar-boxylic acid cycle and metabolic pathways of serum glutamine, serine, tryptophan, pyrimidine, and pyruvate were the most relevant metabolic pathways involved in reducing oxidative stress levels by Erchen decoction during DPDRS treatment; especially, mitochondrial redox homeostasis maintenance in endothelial cells may be crucial. In this work, the therapeutic potential of Erchen decoction for reducing the oxidative stress level in DPDRS was demonstrated; however, its in-depth mechanism is worth further exploration.

目的:探讨稳心颗粒对心肌梗死大鼠心肌缝隙连接蛋白43表达及自噬水平变化的影响.方法:通过心脏左冠状动脉前降支结扎法构建心肌梗死大鼠模型后,随机分为模型组、稳心颗粒低剂量组、稳心颗粒高剂量组及美托洛尔组,连续给药4周,另设正常组作为空白对照,每组8只大鼠.超声心动图检测各组大鼠左室短轴收缩末期前壁厚度(LVAWs)、左室舒张末期前壁厚度(LVAWd)、左室收缩末期内径(LVIDs)、左室舒张末期内径(LVIDd)以及左室射血分数(LVEF);心脏生理电刺激检测各组大鼠的心室颤动阈值;苏木精-伊红(HE)染色观察心脏组织病理学改变;采用免疫组织化学染色及蛋白免疫印迹法检测心肌缝隙连接蛋白43(CX43)、自噬相关蛋白微管相关蛋白1轻链3(LC3)、酵母自噬相关基因6哺乳动物同源体(Beclin1)、P62蛋白的表达.结果:与正常组比较,模型组大鼠LVAWs、LVAWd、LVEF降低,LVIDs、LVIDd升高(P<0.01);与模型组比较,稳心颗粒低剂量组、稳心颗粒高剂量组、美托洛尔组LVAWs、LVAWd、LVEF升高,LVIDs、LVIDd降低(P<0.05或P<0.01).模型组大鼠心室颤动阈值明显低于正常组(P<0.01);与模型组比较,稳心颗粒低剂量组、稳心颗粒高剂量组、美托洛尔组心室颤动阈值有所增高(P<0.01).与正常组比较,模型组LC3、Becli1蛋白平均光密度值明显增加,CX43蛋白平均光密度明显降低(P<0.01);与模型组比较,稳心颗粒高剂量组、美托洛尔组自噬正相关蛋白LC3、Beclin1平均光密度值均明显降低,CX43蛋白平均光密度值增加(P<0.01).与正常组比较,模型组自噬正相关蛋白LC3Ⅱ/LC3Ⅰ、Beclin1表达增加,自噬负相关蛋白P62表达以及CX43蛋白表达降低(P<0.05);与模型组比较,稳心颗粒低剂量组Beclin1蛋白表达明显减少,P62蛋白表达明显升高(P<0.05),稳心颗粒高剂量组及美托洛尔组LC3Ⅱ、Beclin1蛋白表达明显降低,P62及CX43蛋白表达明显升高(P<0.05).结论:稳心颗粒可改善心肌梗死大鼠的心功能并预防心律失常,其机制与调节心脏自噬水平和缝隙连接蛋白43表达有关.

Background MicroRNAs (miRNAs) are endogenous noncoding single-stranded small RNAs. Numerous studies have shown that miRNAs have pivotal roles in the occurrence and development of myocardial fibrosis (MF). However, miRNA expression profile in rats with MF after myocardial infarction (MI) is not well understood. The present study aimed to find the potential miRNA for MF post MI. Methods SPF male Sprague-Dawley (SD) rat models of acute myocardial infarction (AMI) were established by ligating the anterior descending branch of the left coronary artery, while sham-operated rats were only threaded without ligation as a control group. Hematoxylin-eosin and Masson trichrome staining were used to detect myocardial histopathological changes for model evaluation. The differentially expressed miRNAs were detected by using the Agilent Rat miRNA gene chip in the myocardial tissue of the infarct marginal zone. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed by DAVID. The expression of miR-199a-5p was verified by real-time fluorescence quantitative PCR (qRT-PCR). Transfected miR-199a-5p mimics into cardiac fibroblasts (CFs) to construct cell models of miR-199a-5p overexpression. Dual-luciferase reporter assay was employed to validate the target gene of miR-199a-5p. The protein expression of the target gene in CFs transfected with miR-199a-5p mimics were detected by Western blot. Results Myocardial fibrosis was exacerbated in the model group compared with the control group. Thirteen differentially expressed miRNAs between the two groups were screened and their expression levels in the model group were all higher than those in the control group. The expression of miR-199a-5p was significantly increased in the model group in qRT-PCR, which was consistent with the results of the gene chip. KEGG enrichment analysis showed that the target genes of miR-199a-5p were enriched in the insulin signaling pathway. Furthermore, dual-luciferase reporter assay indicated that miR-199a-5p could negatively regulate the expression of GSK-3β. After transfection, the expression of miR-199a-5p was increased in the miR-199a-5p mimics group. The protein expression of GSK-3β was decreased in CFs transfected with miR-199a-5p mimics. Conclusion Our study identified miR-199a-5p could promote the progression of myocardial fibrosis after myocardial infarction by targeting GSK-3β, which provides novel targets for diagnosis and treatment of MF.

目的:探讨异甘草素通过激活PPAR-γ信号通路调控ox-LDL代谢稳定动脉粥样硬化(AS)斑块的分子机制.方法:采用高脂喂养+右侧颈总动脉外置套管术(PCCP)制备ApoE-/-小鼠AS颈动脉斑块模型.ApoE-/-小鼠经PCCP术后随机分为模型组和异甘草素组,正常组采用C57BL/6J小鼠.术后高脂饲料继续喂养8周,建立AS模型.全自动生化仪检测血清中总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)含量.ELISA检测血清中氧化低密度脂蛋白(ox-LDL)含量.HE染色观察颈动脉病理形态,并测定颈动脉参数.油红O染色用于脂质测定,Masson染色用于胶原含量测定,MOMA-2和α-SMA免疫组织化学染色用于巨噬细胞和平滑肌细胞测定,并计算易损指数.Western blot检测小鼠动脉中PPAR-γ、LXR-α、FABP-4、MMP-2及MMP-9蛋白表达.结果:与正常组相比,模型组TC、TG、LDL-C、HDL-C和ox-LDL升高.与模型组相比,异甘草素组TC、TG、LDL-C和ox-LDL降低,HDL-C无明显变化.与正常组相比,模型组小鼠颈动脉的内膜厚度(IT)、内膜/中膜厚度(IT/MT)、斑块面积(PA)、斑块面积/血管管腔面积(PA/LA)均增加,斑块内脂质和MOMA-2含量增加,胶原和α-SMA含量减少,易损指数较高,PPAR-γ、LXR-α表达减少,FABP-4、MMP-2、MMP-9表达增加.与模型组比较,异甘草素组小鼠颈动脉IT、IT/MT、PA、PA/LA均减少,斑块内脂质和MOMA-2含量减少,胶原和α-SMA含量增加,易损指数降低,PPAR-γ、LXR-α表达增加,FABP-4、MMP-2、MMP-9表达减少.结论:异甘草素能够通过激活PPAR-γ、上调LXR-α,减少FABP-4表达,降低ox-LDL水平,减少MMP-2和MMP-9的蛋白表达,降低斑块易损指数,增加斑块稳定性,发挥抗AS的作用.

目的:探讨黄芪注射液对血管紧张素Ⅱ(AngⅡ)诱导的H9c2心肌细胞凋亡的抑制作用及其可能作用机制.方法:采用细胞计数试剂盒-8(CCK-8)方法筛选H9c2细胞培养的黄芪注射液浓度;H9c2细胞分为正常对照组、AngⅡ模型组、AngⅡ+黄芪注射液组.采用实时定量聚合酶链式反应(RT-PCR)法检测3组H9c2细胞电压依赖性阴离子通道1(VDAC1)、线粒体融合蛋白2(Mfn2)及分子伴侣葡萄糖调节蛋白75(GRP75)mRNA的表达;蛋白免疫印迹法(Western Blot)检测VDAC1、Mfn2、GRP75的蛋白表达;Fura-2 AM法测定细胞内Ca2+浓度变化;用流式细胞仪检测各组细胞凋亡情况.结果:黄芪注射液在浓度为0.125％时对细胞的促增殖作用最为显著.与正常对照组比较,AngⅡ模型组Mfn2、GRP75 mRNA表达水平明显增加(P<0.05),AngⅡ+黄芪注射液组Mfn2、GRP75 mRNA表达低于AngⅡ模型组(P<0.05或P<0.01),而3组VDAC1 mRNA表达比较差异无统计学意义(P>0.05).与正常对照组比较,AngⅡ模型组VDAC1、Mfn2和GRP75的蛋白表达明显增加(P<0.05或P<0.01),AngⅡ+黄芪注射液组VDAC1、Mfn2、GRP75的蛋白表达低于AngⅡ模型组(P<0.01).与正常对照组比较,AngⅡ模型组细胞钙含量、细胞凋亡率明显增加(P<0.05或P<0.01),AngⅡ+黄芪注射液组细胞钙含量、细胞凋亡率低于AngⅡ模型组(P<0.01).结论:黄芪注射液能够抑制血管紧张素Ⅱ诱导的H9c2心肌细胞凋亡,其机制可能与其调节VDAC1、Mfn2和GRP75等MAM结构蛋白表达和影响细胞内钙平衡有关.

Abstract: Atherosclerosis is the primary cause of many cardiovascular diseases, and an increasing number of studies have shown that berberine could delay plaque formation and development. Therefore, we aimed to evaluate its effects and explore its mechanisms in this meta-analysis. We searched PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Wanfang, and VIP databases for original preclinical studies to conduct meta-analysis. Twelve articles (16 studies; 312 ApoE−/− mice) were included, and all the studies scored 3–5 points according to SYRCLE's risk of bias tool. Berberine could significantly decrease plaque area and plaque macrophage content (plaque area, SMD = −2.02, 95% CI: −2.80 to −1.24, P = 0.000; plaque macrophage content, SMD = −4.28, 95% CI: −7.67 to −0.88, P = 0.013); lower the levels of TC, triglyceride, and low-density lipoprotein (TC, SMD = −1.47, 95% CI: −2.20 to −0.74, P = 0.000; triglyceride, SMD = −0.77, 95% CI: −1.21 to −0.33, P = 0.000; low-density lipoprotein, SMD = −0.61, 95% CI: −1.11 to −0.11, P = 0.000), and change the secretion of inflammatory cytokines (IL-1β, SMD = −2.29, 95% CI: −3.40 to −1.18, P = 0.000; interleukin-6, SMD = −1.48, 95% CI: −2.11 to −0.85, P = 0.008; tumor necrosis factor-α, SMD = −1.98, 95% CI: −3.01 to −0.94, P = 0.000; interleukin-10, SMD = 1.78, 95% CI: 0.76 to 2.80, P = 0.015), but there were no significant differences in high-density lipoprotein levels and plaque lipid content (high-density lipoprotein, SMD = 0.02, 95% CI: −0.35 to 0.40, P = 0.021; plaque lipid content, SMD = −6.85, 95% CI: −21.09 to 7.39, P = 0.007). The results were robust across a range of sensitivity analyses. Therefore, the results indicate that berberine is a promising drug for the treatment of atherosclerosis through regulating lipid metabolism, inflammation, and plaque composition. However, some potential mechanisms remain to be further elucidated.

Objective: To explore the intervention effects and mechanism of Simiao Yongan (SMYA) Decoction on carotid atherosclerotic plaque in ApoE knockout mice by antagonizing lipid metabolism of oxidation low density lipoprotein (Ox-LDL). Methods: ApoE-/- mice were fed high-fat feeding for 2 weeks, combined with perivascular collar placement (PCCP) and high fat feeding for 8 weeks to establish carotid AS plaque model. All the mouses were divided into 5 groups, after successful AS model preparation. The sham operation as well as model group intragastrically administered with deionized water. The rest mouses were intragastrically administered with SMYA decoction or Pioglitazone, or treated with Atorvastatin for 8 weeks. Then adopting automatic biochemical analyzer to test total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) content. The content of ox-LDL in serum , which was tested by enzyme-linked immunosorbent assay (ELISA). The pathological changes of carotid artery were observed by hematoxylin-eosin (HE) staining and the vascular parameters were measured. Immunohistochemical method and Western blot assay was used to detect the manifestations of protein include fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor γ (PPARγ) as well as matrix metalloproteinase 2 (MMP 2). Results: Compared with the sham operation group, the serum ox-LDL content of the model group was increased, and the intima thickness (IT), intima-media thickness (IT/MT), plaque area (PA) as well as lumen area ratio of plaque vessels (PA/LA) were enlarged (P < 0.05). Immunohistochemistry and Western blot results showed that the relative expression of FABP4 and MMP2 in the model group increased markedly, while the relative expression of PPARγsignificantly decreased. Compared SMYA decoction group and Atorvastatin group as well as Pioglitazone with model group, the serum ox-LDL content was decreased (P < 0.05). In the mean time, the IT, PA, IT/MT and PA/LA in carotid artery were reduced (P < 0.05). Immunohistochemistry and Western blot results showed that the expression of FABP4 and MMP2 was reduced, while PPARγ protein expression increased of mouses in each administration group. Conclusion: SMYA decoction is able to antagonize Ox-LDL, increase the expression of PPARγ, decrease the level of FABP4, reduce MMP2 in carotid arteries of ApoE-/- atherosclerotic mice, and increase the stability of plaque.

目的 探讨血府逐瘀胶囊治疗动脉粥样硬化(AS)的可能作用机制.方法 30只载脂蛋白E基因敲除(ApoE-/-)小鼠随机分为模型组、血府逐瘀胶囊组、阿托伐他汀组,每组10只,另设10只C57BL/6N小鼠为正常组.除正常组外,其他各组小鼠均给予高脂饲料喂养6周制备AS早期模型.造模成功后血府逐瘀胶囊组给予血府逐瘀汤0.6g/(kg·d)灌胃,阿托伐他汀组给予阿托伐他汀3.3 mg/(kg·d)灌胃,正常组和模型组给予0.3 ml去离子水灌胃,给药期间除正常组外其余各组继续给予高脂饲料喂养.干预12周后观察小鼠主动脉瓣病理形态学改变和脂质沉积情况;检测血脂指标[包括血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)],并计算TC/HDL-C、LDL-C/HDL-C和动脉粥样硬化指数(AI);检测小鼠主动脉整合素相关蛋白分子(CD47)、过氧化物酶体增殖物激活受体γ(PPAR-γ)、肝X受体α(NR1H3)蛋白表达水平.结果 与正常组比较,模型组小鼠主动脉瓣斑块面积和脂质沉积增多,血清TC、LDL-C、TC/HDL-C、LDL-C/HDL-C和AI水平升高,主动脉CD47蛋白表达升高,HDL-C水平降低(P＜0.01).与模型组相比,血府逐瘀胶囊组和阿托伐他汀组主动脉瓣斑块面积和脂质沉积减少,小鼠血清TC、LDL-C水平及CD47蛋白表达下降,NR1H3蛋白表达升高(P＜0.01),血府逐瘀胶囊组PPAR-γ蛋白表达水平升高(P＜0.01);阿托伐他汀组HDL-C水平升高,TC/HDL-C、LDL-C/HDL-C和AI显著降低(P＜0.05或P＜0.01).与阿托伐他汀组相比,血府逐瘀胶囊组CD47蛋白表达水平升高(P＜0.05).结论 血府逐瘀胶囊治疗AS可能与调控炎症关键分子CD47、PPAR-γ、NR1H3的表达有关.

Aims: The study aimed to evaluate the correlation of different microparticle (MP) phenotypes with plaque burden and their diagnostic value and preliminarily explore the role of MPs in atherosclerosis (AS). Methods: Carotid intima-media thickness (CIMT) and maximal plaque area in 23 patients with carotid atherosclerosis (CAS) and 22 healthy subjects were measured by ultrasound. Transmission electron microscopy, nanoparticle tracking analysis and western blot were used to identify MPs. Flow cytometry assay measured absolute number of MPs, and receiver operating characteristic (ROC) analysis was used to assess the relationship between plaque burden and MPs. To study the preliminary mechanism of MPs in AS, MPs were administered to 32 male Kunming mice, which were randomly divided into control, CAS, healthy, and tetrahydrobiopterin (BH4) groups. Hematoxylin-eosin staining, immunohistochemistry staining, and Western blot were adopted to detect relevant indexes 24 h after the injection. Results: The plasma levels of CD45+ leukocyte-derived microparticle (LMP), CD11a+ LMP, CD11a+/CD45+ LMP, and CD31+/CD42b+ platelet-derived microparticle (PMP) in CAS patients were significantly higher than those in healthy subjects, and were positively correlated with the maximal plaque area. Moreover, the levels of CD11a+ LMP, CD11a+/CD45+ LMP were also positively correlated with CIMT. The area under the ROC curve of the four MPs was 0.689, 0.747, 0.741, and 0.701, respectively. Compared with healthy subjects, MPs from CAS patients resulted in a significantly lower expression of endothelial nitric oxide synthase (eNOS) dimer/monomer, and BH4 could improve eNOS uncoupling. Moreover, the level of VCAM-1 in intima in the CAS group was significantly higher than in the other three groups. Conclusion: CD11a+ LMP and CD11a+/CD45+ LMP might be potential biomarkers for CAS prediction. BH4-related eNOS uncoupling occurs in CAS patients, and circulating MPs from them lead to endothelial dysfunction through eNOS uncoupling.

Simiao Yong’an decoction (SMYAD), a classic traditional Chinese medicine formula, has been used to treat atherosclerosis (AS) in clinical in China, but its therapeutic mechanism and pharmacodynamic material basis are not clear. In this study, the AS model was caused by a high-fat diet and perivascular carotid collar placement (PCCP), and SMYAD was orally administered to the model and normal mice. A rapid, sensitive, selective, and reliable method using ultrahigh-performance liquid chromatography (UHPLC) system combined with a Q Exactive HF-X mass spectrometer (UHPLC-Q Exactive HF-X MS) was established and validated for the simultaneous determination of seven compounds, including harpagide, chlorogenic acid, swertiamarin, sweroside, angoroside C, liquiritin, and isoliquiritigenin in the plasma of normal and AS mice. The specificity, linearity, precision, accuracy, recovery, and stability of the method were all within the acceptable criteria. The results showed that some pharmacokinetic behaviors of harpagide, chlorogenic acid, and isoliquiritigenin were significantly different among the two groups of mice. The specific parameter changes were harpagide (AUC0–t and AUC0–∞ were 11075.09 ± 2132.38 and 16221.95 ± 5622.42 ng·mL−1·h, respectively; CLz/F was 2.45 ± 0.87 L/h/mg), chlorogenic acid (t1/2 was 21.59 ± 9.16 h; AUC0–∞ was 2637.51 ± 322.54 ng·mL−1·h; CLz/F was 13.49 ± 1.81 L/h/mg) and isoliquiritigenin (AUC0–t and AUC0–∞ were 502.25 ± 165.65 and 653.68 ± 251.34 ng·mL−1·h, respectively; CLz/F was 62.16 ± 23.35 L/h/mg) were altered under the pathological status of AS. These differences might be partly ascribed to the changes in gastrointestinal microbiota, nonspecific drug transporters, and cytochrome P450 activity under the AS state, providing research ideas and experimental basis for pharmacological effects and pharmacodynamic material basis.

Rosiglitazone (Avandia) and pioglitazone (Actos) belong to the class of thiazolidinediones (TZDs) drugs that act by increasing insulin sensitivity and are widely used for treating diabetic patients with insulin resistance. TZDs exhibit anti-inflammatory and antioxidant properties, then may play an active role in inhibiting plaque formation and coronary atherosclerosis. But the results of evidence-based medicine suggest that TZDs may increase the risk of cardiovascular adverse events. To explore the dispute in depth, our meta-analysis aimed to evaluate the changes in vascular endothelial and plaque-related indicators following treatment with TZDs in diabetic patients with coronary atherosclerosis. According to our meta-analysis, TZDs showed an inhibiting effect on plaque progression and a protective effect on the vascular endothelium in patients with diabetes and coronary atherosclerosis. Interestingly, these effects may not depend on the regulation of inflammation and lipid metabolism. By this token, TZDs may develop a potential protective effect on myocardial infarction. Systematic review registration [https://www.crd.york.ac.uk/prospero/], identifier [CRD42021231663].

Background. Myocardial infarction (MI) is an acute and serious cardiovascular disease. Arrhythmia after MI can lead to sudden cardiac death, which seriously affects the survival outcome of patients. WenXin KeLi is a Chinese patent medicine for the treatment of arrhythmia in a clinic, which can significantly improve symptoms of palpitation and play an important role in reducing the risk of arrhythmia after MI. In this study, we aimed to explore the pharmacological mechanism of WenXin KeLi in protecting the heart. Methods. The MI model was established by ligating the left coronary artery and the ventricular fibrillation threshold (VFT) was measured by electrical stimulation. The expression of connexin43 (CX43) and autophagy-related protein were measured by Western Blot, and correlation analysis was conducted to study the relationship between cardiac autophagy, CX43, and arrhythmia in rats after MI. The effects of WenXin KeLi on arrhythmia, cardiac structure, and function in MI rats were respectively observed by electrical stimulation, cardiac gross section, Masson staining, and cardiac ultrasound. The effects of WenXin KeLi on the expression of phosphoinositide 3 kinase-protein kinase B-mammalian targets of rapamycin (PI3K-AKT-mTOR) autophagy pathway and CX43 were observed by Western Blot. Results. After 4 weeks of MI, the VFT in the model group was significantly reduced, the expression levels of yeast ATG6 homolog (Beclin1), microtubule-associated protein 1A/1B-light chain 3 (LC3II/LC3I), and p-CX43 (S368) significantly increased, the expression of sequestosome-1(P62) and CX43 significantly decreased. LC3II/LC3I and Beclin1 expression were significantly negatively correlated with the VFT, and the expression of P62 and CX43 were significantly positively correlated with the VFT. LC3II/LC3I and Beclin1 expression were negatively correlated with CX43 expression, while P62 expression was positively correlated with CX43 expression. WenXin KeLi could significantly increase the VFT, reduce the deposition of collagen fibers, and increase the index levels of the left ventricular end-diastolic anterior wall (LVEDAW), interventricular septum end-diastolic (IVSED), left ventricular end-systolic anterior wall (LVESAW), interventricular septum end-systolic (IVSES), left ventricular end-diastolic posterior wall (LVEDPW), left ventricular end-systolic posterior wall (LVESPW), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), and reduce the index levels of the left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic dimension (LVESD), left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV). WenXin KeLi could increase the expression of CX43, P62, AKT, p-PI3K, p-AKT (308), p-AKT (473), and p-mTOR and decrease the expression of LC3II/LC3I and Beclin1. Conclusion. WenXin KeLi can activate the PI3K-AKT-mTOR signaling pathway, improve cardiac autophagy and Cx43 expression in rats after MI, reduce the risk of arrhythmia after MI, and play a cardioprotective role.

目的:比较酒制豨莶草和生豨莶草对大鼠心肌细胞缺氧损伤的保护作用,并探讨其作用机制,为临床用药选择提供实验依据.方法:采用H9 c2细胞缺氧模型,给予不同浓度酒制豨莶草和生豨莶草干预,CCK-8法测定细胞成活率,比色法检测细胞培养上清液中乳酸脱氢酶(LDH)活性,超氧化物歧化酶(SOD)活性;荧光酶标仪检测细胞内活性氧(ROS)含量;用蛋白质印迹法(Western Blotting)检测凋亡B细胞淋巴瘤-2(Bcl-2)蛋白和Bcl-2相关X蛋白(Bax)的表达.结果:缺氧16 h成功制备H9 c2心肌细胞缺氧损伤模型;筛选确定300μg/mL是生豨莶草和酒制豨莶草最佳给药浓度;与模型组比较,酒制豨莶草和生豨莶草均可提高细胞成活率(P<0.05),改善损伤细胞形态,减轻LDH外漏(P<0.05),降低ROS的含量(P<0.05),增加SOD活性(P<0.05),上调Bcl2(P<0.05)和Bcl-2/Bax的表达.给药浓度为300μg/mL时,生豨莶草在提高SOD活力和细胞成活率方面优于酒制豨莶草(P<0.05),酒制豨莶草在抑制LDH外漏的作用强于生豨莶草,在活性氧(ROS)含量和Bcl、Bax蛋白表达方面,差异无统计学意义(P>0.05).结论:酒制豨莶草和生豨莶草对缺氧损伤的H9 c2心肌细胞具有保护作用,其机制可能是通过抵抗氧化应激,抑制心肌细胞凋亡.

Objective: To explore the cardioprotective mechanism of Wenxin Granules regulating the expression of apoptosis-related genes in cardiomyocytes. Methods: A rat model of myocardial infarction was established and randomly divided into model group, Wenxin granule low-dose group, Wenxin granule high-dose group, metoprolol group and sham operation group. the left ventricular end systole anterior wall thickness(LVAWs), end systole inner diameter (LVIDs), end systole posterior wall thickness (LVPWs), end-diastolic anterior wall thickness (LVAWd), end-diastolic inner diameter (LVIDd), end-diastolic posterior wall thickness(LVPWd)and left ventricular ejection fraction (LVEF) were detected by echocardiography in each group after 2 weeks of treatment. Hematoxylin eosin (HE) staining was used to observe the changes in the cardiac structure of rats in each group. Real-time PCR (Real-time PCR) was used to detect the relative expression of mammalian B-cell lymphoma-2 (BCL-2), BCL-2 related X protein (BAX), Caspase-9 (Caspase-9), and Caspase-3 (Caspase-3) mRNA. TUNEL staining was used to detect changes in the apoptotic rate of rat cardiomyocytes in each group. Results: Compared with the sham operation group, the LVAWs, LVPWs, LVPWd and LVEF of the model group were significantly reduced (P<0.05, P<0.01), and LVIDs and LVIDd were significantly increased (P<0.05, P<0.01). Severe pathological ischemia injury of heart tissue. The relative expression of BCL-2 mRNA and the ratio of BCL-2/BAX in the model group were significantly reduced (P<0.01), while the relative expression of BAX, Caspase-9 and Caspase-3 mRNA was significantly increased (P<0.01). The apoptosis rate was significantly increased (P<0.01). In the low-dose and high-dose groups of Wenxin Granules and the Metoprolol group, LVAWs, LVPWs, LVPW d, and LVEF of rats in each administration group increased significantly (P<0.05, P<0.01), LVIDs , LVIDd was significantly reduced (P<0.05, P<0.01), the pathological damage of the heart tissue was improved, the expression of BCL-2 mRNA and the ratio of BCL-2/BAX were significantly increased (P<0.05, P<0.01), BAX, The expression of Caspase-9 and Caspase-3 mRNA was significantly reduced (P<0.05, P<0.01), and the apoptotic rate of myocardial cells was significantly reduced (P<0.01). Conclusion: Wenxin granule can play a cardioprotective role by regulating the gene expression of BCL-2/BAX/Caspase apoptosis pathway.

Abstract Rosiglitazone (Avandia) and Pioglitazone (Actos) belong to the class of thiazolidinedione drugs that act by increasing insulin sensitivity and are widely used for treating diabetic patients with insulin resistance. Thiazolidinediones (TZDs) exhibit anti-inflammatory and antioxidant properties and inhibit endothelial cell inflammation and dysfunction, and then play a role in inhibiting plaque formation and coronary atherosclerosis. But the results of evidence-based medicine suggest that thiazolinedione may increase the risk of myocardial infarction. To explore the dispute in depth, the meta-analysis aimed to evaluate the changes in vascular endothelial and plaque-related indicators following treatment with thiazolidinediones on diabetic patients with coronary atherosclerosis.According to our meta-analysis, Thiazolinediones showed a protective effect on the vascular endothelium and an inhibiting effect on plaque progression in patients with diabetes and coronary atherosclerosis and these effects may not depend on related targets or pathways such as inflammation and lipid regulation. Due to the poor quality of the evidence, more and higher-quality studies are needed to further improve the above conclusions.

BackgroundUnlike chemical drugs with a single or a few kinds of active compounds, traditional Chinese medicines (TCMs)uses herbal formulas composed of numerous kinds of chemical constituents. Therefore, TCM clinical trials require unique and stricter standards for collecting, preserving, and transporting fecal samples than those used for chemical drugs. Unfortunately, there are no special standards for processing fecal samples in TCM clinical trials.MethodsWe invited interdisciplinary experts within TCM clinical trials and gut microbiome research to help formulate this standard. After more than a year’s in-depth discussion and amendments, we achieved a standard via expert interviews, literature research, questionnaire surveys, and public opinion solicitation. This standard has been reviewed and approved by the Standards Office of China of the Association of Chinese medicine.ResultsWe established a sample information processing method prior to TCM clinical sample collection, which is adapted to the unique features of TCM. The method formulates detailed processing requirements for TCM information in addition to the factors that may disturb the gut microbiome. We also constructed a set of methods for collecting, preserving, and transporting fecal samples that meet the characteristics of TCM. These methods formulate detailed operating specifications on the collection approaches, storage conditions, transportation requirements, and management of fecal samples.ConclusionsThis standard guides the information processing prior to sample collection and the standard operating procedures for the collection, preservation, and transportation of fecal samples in TCM clinical trials, which also can be used as a reference by clinicians and researchers in modern medicines.

目的:探讨稳心颗粒调节缝隙连接蛋白43(CX43)和内质网应激通路基因表达的心脏保护机制.方法:建立心肌梗死大鼠模型,术后第2天根据心电图筛选入组,随机分为模型组、稳心颗粒低剂量组、稳心颗粒高剂量组、美托洛尔组,另设平行操作不结扎的假手术组,药物治疗2周.腹腔注射异丙肾上腺素记录心律失常发生率;心脏血流动力学检测各组大鼠左心室收缩压(LVSP)、左心室舒张末压(LVEDP)、左心室内压最大上升速率(+dp/dtmax)、左心室内压最大下降速率(–dp/dtmax);苏木素-伊红(HE)染色观察各组大鼠心脏病理改变;Masson染色检测心肌梗死边缘区胶原容积分数;实时荧光定量聚合酶链式反应(qRT-PCR)检测各组大鼠CX43、葡萄糖调节蛋白78(GRP78)、肌醇需求激酶1(IRE1)、活化转录因子6(ATF6)、X盒结合蛋白1(XBP1)mRNA的相对表达量;蛋白免疫印迹法检测CX43的表达.结果:与假手术组相比,模型组大鼠心律失常发生率明显上升(P<0.01),LVSP、+dp/dtmax、–dp/dtmax均显著降低(P<0.01),LVEDP显著增高(P<0.01),心肌组织病理性改变明显,心肌胶原容积分数明显增加(P<0.01),CX43 mRNA相对表达量明显下调(P<0.01),IRE1 mRNA相对表达量增高(P<0.05),GRP78、ATF6、XBP1 mRNA相对表达量明显增高(P<0.01),CX43蛋白表达明显降低(P<0.01);稳心颗粒及美托洛尔治疗2周后,给药组大鼠心律失常发生率下降(P<0.05),LVSP增加(P<0.05),+dp/dtmax、–dp/dtmax显著增加(P<0.01),心肌组织病理性变化显著改善,心肌胶原容积分数明显降低(P<0.01),CX43 mRNA表达量上调(P<0.05),GRP78、IRE1、ATF6、XBP1 mRNA表达量下调(P<0.05),CX43蛋白表达升高(P<0.05).结论:稳心颗粒可调节心肌梗死大鼠CX43和内质网应激通路基因表达,进而发挥心脏保护作用.