Vitamin D receptor (VDR) mediates the quiescence of activated hepatic stellate cells (aHSCs) by regulating M2 macrophage exosomal smooth muscle cell-associated protein 5 (SMAP-5) / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism. Hepatic fibrosis is characterized by activated hepatic stellate cells (aHSCs) with an excessive production of extracellular matrix. Although promoted activation of HSCs by M2 macrophages has been demonstrated, the molecular mechanism involved remains ambiguous. Herein, we propose that the vitamin D receptor (VDR) involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes. We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation. The exosomes derived from M2 macrophages can promote HSC activation, while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes. Smooth muscle cell-associated protein 5 (SMAP-5) was found to be the key effector protein in promoting HSC activation by regulating autophagy flux. Building on these results, we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect. In this study, we aim to elucidate the association between VDR and macrophages in HSC activation. The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis, and provide potential therapeutic targets for its treatment.

Protective mechanism of salvianolic acid B on blood vessels / 中国中药杂志

Salvianolic acid B(Sal B) is the main water-soluble component of Salvia miltiorrhiza Bunge. Studies have found that Sal B has a good protective effect on blood vessels. Sal B can protect endothelial cells by anti-oxidative stress, inducing autophagy, inhibiting endoplasmic reticulum stress(ERS), inhibiting endothelial inflammation and adhesion molecule expression, inhibiting endothelial cell permeability, anti-thrombosis, and other ways. In addition, Sal B can alleviate endothelial cell damage caused by high glucose(HG). For vascular smooth muscle cell(VSMC), Sal B can reduce the synthesis and secretion of inflammatory factors by inhibiting cyclooxygenase. It can also play a vasodilatory role by inhibiting Ca~(2+) influx. In addition, Sal B can inhibit VSMC proliferation and migration, thereby alleviating vascular stenosis. Sal B also inhibits lipid deposition in the subendothelium, inhibits macrophage conversion to foam cells, and reduces macrophage apoptosis, thereby reducing the volume of subendothelial lipid plaques. For some atherosclerosis(AS) complications, such as peripheral artery disease(PAD), Sal B can promote angiogenesis, thereby improving ischemia. It should be pointed out that the conclusions obtained from different experiments are not completely consistent, which needs further research. In addition, previous pharmacokinetics showed that Sal B was poorly absorbed by oral administration, and it was unstable in the stomach, with a large first-pass effect in the liver. Sal B had fast distribution and metabolism in vivo and short drug action time. These affect the bioavailability and biological effects of Sal B, and the development of clinically valuable Sal B non-injectable delivery systems remains a great challenge.

Preliminary Results of Single-cell Transcriptome Sequencing in Renal Arterial Lesions of Takayasu Arteritis / 中国医学科学院学报

Objective To explore the preliminary application of single-cell RNA sequencing (scRNA-seq) in the renal arterial lesions in Takayasu arteritis (TA) patients. Methods This study included 2 TA patients with renal artery stenosis treated by bypass surgery in the Department of Vascular Surgery,Beijing Hospital.The obtained 2 renal artery samples were digested with two different protocols (GEXSCOPE kit and self-made digestion liquid) before scRNA-seq and bioinformatics analysis. Results A total of 2920 cells were obtained for further analysis.After unbiased cluster analysis,2 endothelial cell subsets,2 smooth muscle cell subsets,1 fibroblast subset,2 mononuclear macrophage subsets,1 T cell subset,and 1 undefined cell subset were identified.Among them,the two subsets of smooth muscle cells were contractile and secretory,respectively.The results of scRNA-seq indicated that enzymatic hydrolysis with GEXSCOPE kit produced a large number of endothelial cells (57.46%) and a small number of immune cells (13.21%).However,immune cells (34.64%) were dominant in the cells obtained by enzymatic hydrolysis with self-made digestive liquid. Conclusion scRNA-seq can be employed to explore the cellular heterogeneity of diseased vessels in TA patients.Different enzymatic digestion protocols may impact the proportion of different cells.

A nonobstructive condition: Medial arterial calcification / 中南大学学报(医学版)

Vascular calcification, including intimal and medial calcification, is closely associated with a significant increase in cardiovascular diseases. Although increased understandings were achieved, people still know much more about intimal calcification than medial calcification because the latter doesn't obstruct the arterial lumen, commonly considered as a non-significant finding. We clarified the pathologic characteristic of medial calcification, its difference from intimal calcification, principally focused on its clinical relevance, such as diagnosis, nosogenesis, and hemodynamics. We underline the importance of identifying and distinguishing medial calcification, understanding its effect to local/systematic arterial compliance, and relationship to diabetic neuropathy. Recent studies emphasize do not ignore its predictive role in cardiovascular mortality. It is of great clinical significance to summarize the mechanisms of occurrence, lesion characteristics, diagnostic methods, pathogenic mechanisms, hemodynamic changes, and the distinction as well as association of intimal calcification with intimal calcification.

Terpine-4-ol on inhibiting vascular smooth muscle cell calcification by improving mitochondrial dynamics / 药学学报

The aim of this study was to investigate the role and mechanism of terpinen-4-ol (T4O) on high glucose (HG) -induced calcification in vascular smooth muscle cell (VSMC). To investigate the role of T4O on HG-induced calcium deposition, osteogenic phenotypic transformation and mitochondrial dynamics in VSMC, Mdivi-1, a mitochondrial dynamin-related protein 1 (Drp-1) inhibitor, was used to analyze the correlation between mitochondrial dynamics and VSMC calcification and the role of T4O. Alizarin red S staining was used to observe calcium salt deposition and flow cytometry to detect intracellular Ca2+ content; Western blot and immunofluorescence were used to detect the expression of phenotypic switching-related markers α-smooth muscle actin (α-SMA), bone morphogenetic protein 2 (BMP2) and Runt related transcription factor 2 (Runx2), and mitochondrial dynamics-related markers mitofusin 1 (MFN1), mitofusin 2 (MFN2) and Drp-1. The results showed that low and high doses of T4O could inhibit HG-induced down-regulation of α-SMA, MFN1 and MFN2 expression levels, and up-regulation of BMP2, Runx2 and Drp-1 expression levels, reduce intracellular Ca2+ content and calcium salt deposition, and effectively inhibit HG-induced VSMC calcification and mitochondrial dynamics disorders. The T4O group, Mdivi-1 group and T4O+Mdivi-1 group were able to up-regulate the expression levels of HG-induced α-SMA, MFN1 and MFN2, down-regulate the protein expression levels of BMP2, Runx2 and Drp-1, and inhibit calcium salt deposition, and there was no significant difference between the above indexes in the T4O and T4O+Mdivi-1 groups. The above findings suggest that T4O can inhibit the expression level of Drp-1, regulate the disturbance of mitochondrial dynamics, and suppress HG-induced VSMC calcification.

The substitution of SERCA2 redox cysteine 674 promotes pulmonary vascular remodeling by activating IRE1α/XBP1s pathway

Pulmonary hypertension (PH) is a life-threatening disease characterized by pulmonary vascular remodeling, in which hyperproliferation of pulmonary artery smooth muscle cells (PASMCs) plays an important role. The cysteine 674 (C674) in the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2 (SERCA2) is the critical redox regulatory cysteine to regulate SERCA2 activity. Heterozygous SERCA2 C674S knock-in mice (SKI), where one copy of C674 was substituted by serine to represent partial C674 oxidative inactivation, developed significant pulmonary vascular remodeling resembling human PH, and their right ventricular systolic pressure modestly increased with age. In PASMCs, substitution of C674 activated inositol requiring enzyme 1 alpha (IRE1α) and spliced X-box binding protein 1 (XBP1s) pathway, accelerated cell cycle and cell proliferation, which reversed by IRE1α/XBP1s pathway inhibitor 4μ8C. In addition, suppressing the IRE1α/XBP1s pathway prevented pulmonary vascular remodeling caused by substitution of C674. Similar to SERCA2a, SERCA2b is also important to restrict the proliferation of PASMCs. Our study articulates the causal effect of C674 oxidative inactivation on the development of pulmonary vascular remodeling and PH, emphasizing the importance of C674 in restricting PASMC proliferation to maintain pulmonary vascular homeostasis. Moreover, the IRE1α/XBP1s pathway and SERCA2 might be potential targets for PH therapy.

Role of nicotinamide phosphoribosyltransferase in delaying smooth muscle cell senescence and protecting abdominal aortic aneurysm / 中华危重病急救医学

Objective:To investigate the protective effect of nicotinamide phosphoribosyltransferase (NAMPT) on abdominal aortic aneurysm by delaying the senescence of aortic vascular smooth muscle cells (VSMC).Methods:The primary VSMC cells from normal and patients with abdominal aortic aneurysm were cultured by tissue adherence method. Cells were divided into normal human-derived VSMC group (Ctrl-VSMC group), abdominal aortic aneurysm patient-derived VSMC group (AAA-VSMC group), and angiotensinⅡ(AngⅡ) in vitro abdominal aortic aneurysm model group (AngⅡ-VSMC group, 100 nmol/L AngⅡ treated normal human-derived VSMC for 48 hours), AngⅡ+P7C3 group and AAA+P7C3 group after NAMPT agonist P7C3 intervention (adding 5 μmol/L P7C3 on the basis of AngⅡ-VSMC group and AAA-VSMC group, respectively). Immunofluorescence staining was used to identify VSMC; cell proliferation-associated antigen Ki67 staining was used to detect cell proliferation; senescence associated β-galactosidase (SA-β-gal) staining was used to detect cell senescence in each group; Western blotting was used to detect the protein expression levels of senescence-related proteins p21, p16 and NAMPT in each group. Results:Compared with the Ctrl-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA-VSMC group and AngⅡ-VSMC group were significantly increased [SA-β-gal staining positive rate: (74.1±4.4)%, (68.6±5.5)% vs. (36.8±10.3)%, p21/GAPDH: 0.61±0.07, 0.51±0.03 vs. 0.31±0.03, p16/GAPDH: 0.77±0.03, 0.72±0.06 vs. 0.33±0.26, all P < 0.01]. However, the expression of NAMPT was significantly decreased (NAMPT/GAPDH: 0.88±0.07, 0.79±0.14 vs. 1.29±0.02, both P < 0.01). Compared with the AngⅡ-VSMC group, the positive rate of SA-β-gal staining and the expressions levels of senescence-related proteins p21 and p16 in the AngⅡ+P7C3 group were significantly lower [SA-β-gal staining positive rate: (49.1±3.2)% vs. (68.6±5.5)%, p21/GAPDH: 0.35±0.06 vs. 0.51±0.03, p16/GAPDH: 0.47±0.08 vs. 0.72±0.06, all P < 0.05], while the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.15±0.06 vs. 0.79±0.14, P < 0.01). Compared with the AAA-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA+P7C3 group were significantly lower [SA-β-gal staining positive rate: (54.1±6.0)% vs. (74.1±4.4)%, p21/GAPDH: 0.38±0.02 vs. 0.61±0.07, p16/GAPDH: 0.50±0.13 vs. 0.77±0.03, all P < 0.05], but the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.25±0.28 vs. 0.88±0.07, P < 0.01). Conclusion:NAMPT agonist P7C3 can delay the senescence of VSMC and play a protective role in abdominal aortic aneurysm.

Treatment of pulmonary hypertension based on inhibition of pulmonary vascular cell proliferation / 药学学报

Pulmonary hypertension is a rapidly progressing disease of the lung vasculature with poor prognosis, ultimately leading to right heart failure and death. The remodeling of small pulmonary arteries represents an important pathological characteristic of pulmonary hypertension. Pulmonary arterial smooth muscle cells (PASMCs) located in the middle layer of pulmonary artery exhibit hyperproliferation and resistance to apoptosis, which is the main initiator of pulmonary vascular remodeling and similar to that seen in tumor cells. In this review we focus on the signaling pathways that play a key role in PASMCs proliferation and the latest research progress on inhibitors targeting cell proliferation pathways to provide a new perspective for the treatment of PH．

Research advance of Nrf2 on atherosclerosis by regulating vascular smooth muscle cell / 浙江大学学报·医学版

Atherosclerosis is a common pathological change in cardiovascular disease. Vascular smooth muscle cell is the main source of plaque cell and extracellular matrix, and nuclear factor-erythroid 2-related factor 2 (Nrf2) is a key transcription factor regulating the function of vascular smooth muscle cell. In the process of atherosclerosis, Nrf2 signaling pathway has the following regulatory effects on vascular smooth muscle cell: regulating the phenotype of vascular smooth muscle cell to change to the direction conducive to the alleviation of disease progression; inhibiting the proliferation and migration of vascular smooth muscle cell; mitigating the level of blood lipid; alleviating vascular smooth muscle cell calcification, aging and apoptosis process. This article reviews the specific mechanisms of Nrf2 regulating atherosclerosis, such as phenotypic transformation, proliferation and migration, lipid metabolism, calcification, aging and apoptosis in atherosclerosis, in order to provide a basis for understanding the molecular mechanism of atherosclerosis development and finding therapeutic targets.

Functional role and underlying mechanisms of neuropilin-1 in proliferation and migration of rat vascular smooth muscle cells / 中国胸心血管外科临床杂志

@#Objective    To investigate the role and potential mechanisms of neuropilin-1 (NRP1) in the pathogenesis of vein graft failure. Methods    The rat vascular smooth muscle cells (VSMCs) were transfected with NRP1-shRNA adenovirus and negative control adenovirus respectively. Cell counting kit-8, flow cytometry, Transwell and Western blot were used to investigate the effects of inhibition of NRP1 on VSMCs proliferation viability, apoptosis, migration capacity and its downstream signaling pathway protein expression. Results    The proliferation and migration of rat VSMCs could be inhibited after down-regulation of NRP1, and the increase of apoptosis was also observed. Moreover, inhibition of NRP1 significantly reduced Akt and NF-κB phosphorylation in rat VSMCs, but had little effect on activation of ERK1/2. Conclusion    NRP1 may promote vein graft hyperplastic remodeling by regulating the proliferation and migration of VSMCs through PI3K/Akt and NF-κB pathways, but further animal study is required.

Research progress on correlation between lncRNAs and pathogenesis of vascular diseases / 中国职业医学

Occupational hand-arm vibration diseases(HAVD) is a legitimate occupational disease in China, and the mechanism of its pathogenesis is vibration-induced vascular injury. Once HAVD occurs, it is difficult for the patients to recover and can cause great harm to workers exposed to hand-arm vibration. It is difficult to detect and evaluate the occurrence and progress of the disease at an early stage using existing technology, which is disadvantageous to the early prevention and treatment of the disease. Long noncoding RNAs(lncRNAs) play an important role in regulating the development, growth, and remodeling of blood vessels and other biological processes. This article reviews the role and mechanism of lncRNAs in vascular injury, and provides scientific theoretical basis for early diagnosis and treatment of HAVD.

Effects of notoginsenoside R1 on AngⅡ induced proliferation of MOVAS cells and AT1R/MAPKs signaling pathway / 上海预防医学

Objective:To investigate the effects of notoginsenoside R1 （NR1） on the proliferation of mice aortic smooth muscle cells （MOVAS cells） induced by angiotensinⅡ （AngⅡ） and the signal pathway of angiotensin Ⅱ type 1 receptor （AT1R） / mitogen activated protein kinases （MAPKs）. Methods:The proliferation of MOVAS cells was detected by BrdU method after AngⅡ induction. Western blot was used to detect the expression of the two main receptors of AngⅡ （AT1R and AT2R） and MAPKs pathway related proteins （ERK， p38， and JNK）. Results:（1） AngⅡ （5 μmol/L） could promote the proliferation of MOVAS cells （P<0.01）. NR1 （50 μmol/L） could inhibit the proliferation of MOVAS cells induced by AngⅡ （P<0.01）. There was no significant difference between control group and NR1 group （P>0.05）. （2） Compared with AngⅡ group， the expression of AT1R protein in AngⅡ+ NR1 group was significantly lower （P<0.05）， but there was no difference in the expression of AT2R protein （P>0.05）. （3） NR1 could significantly inhibit the phosphorylation of ERK， p38 and JNK protein after AngⅡ stimulation （P<0.01）. Conclusion:NR1 can inhibit the proliferation of MOVAS cells induced by AngⅡ， which may be related to down regulating AT1R and inhibiting the activation of MAPKs.

Quercetin-induced apoptosis ameliorates vascular smooth muscle cell senescence through AMP-activated protein kinase signaling pathway

Aging is one of the risk factors for the development of cardiovascular diseases. During the progression of cellular senescence, cells enter a state of irreversible growth arrest and display resistance to apoptosis. As a flavonoid, quercetin induces apoptosis in various cells. Accordingly, we investigated the relationship between quercetin-induced apoptosis and the inhibition of cellular senescence, and determined the mechanism of oxidative stress-induced vascular smooth muscle cell (VSMC) senescence. In cultured VSMCs, hydrogen peroxide (H₂O₂) dose-dependently induced senescence, which was associated with increased numbers of senescence-associated β-galactosidase-positive cells, decreased expression of SMP30, and activation of p53-p21 and p16 pathways. Along with senescence, expression of the anti-apoptotic protein Bcl-2 was observed to increase and the levels of proteins related to the apoptosis pathway were observed to decrease. Quercetin induced apoptosis through the activation of AMP-activated protein kinase. This action led to the alleviation of oxidative stress-induced VSMC senescence. Furthermore, the inhibition of AMPK activation with compound C and siRNA inhibited apoptosis and aggravated VSMC senescence by reversing p53-p21 and p16 pathways. These results suggest that senescent VSMCs are resistant to apoptosis and quercetin-induced apoptosis attenuated the oxidative stress-induced senescence through activation of AMPK. Therefore, induction of apoptosis by polyphenols such as quercetin may be worthy of attention for its anti-aging effects.

Therapeutic effects of albendazole on mice with artery stenosis and its mechanism / 上海交通大学学报（医学版）

Objective • To investigate the effect of albendazole (ABZ) on mouse femoral arteries restenosis and explore its possible mechanism. Methods • Vascular smooth muscle cells (VSMCs) were cultured in vitro with 0.5 and 1 μmol/L ABZ, and evaluated for cell proliferation, migration, and apoptosis by MTT, Transwell assay, and Annexin V-PI staining flow cytometry, respectively; and Western blotting was also used for the analysis of phosphorylation mechanism of cytoskeleton proteins cofilin (CFL) and myosin light chain (MLC). Stenosis was established in the unilateral femoral artery of 10-week-old wildtype male C57BL/6 mice by perivascular cuff placement and high fat chow breeding for 4 weeks. After successful modeling, mice were randomly divided into control group (equal volume of solvent) and ABZ group (1.5 mg/d) for gavage, and femoral arteries were collected 4 weeks later for H-E histological analysis of intimal area, medial area, and intima/media (I/M) ratio to clarify the severity of restenosis. Results • Both 0.5 and 1 μmol/L ABZ could significantly inhibit the proliferation and migration of VSMCs (both P<0.05), while 0.5 μmol/L had no significant effect on the apoptosis of VSMCs. ABZ gavage could significantly reduce the neointimal area and I/M ratio in the restenosis mice, while there were no effects on the median area. Both 0.5 and 1 μmol/L ABZ treatment could significantly inhibit CFL dephosphorylation and MLC phosphorylation, which showed a concentration-dependent trend (both P<0.05). Conclusion • ABZ inhibits VSMCs migration and intimal hyperplasia, via affecting the phosphorylation of cytoskeleton protein CFL and MLC, thereby resulting in therapeutic effects on restenosis of mice.

Effect of Korean Magnolia obovata Extract on Platelet-Derived Growth Factor-Induced Vascular Smooth Muscle Cells / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Korean Magnolia obovata crude extract (KME) on plateletderived growth factor (PDGF)-BB-induced proliferation and migration of vascular smooth muscle cells (VSMCs).@*METHODS@#KME composition was analyzed by high-performance liquid chromatography (HPLC). VSMCs were isolated from the aorta of a Sprague-Dawley rat, incubated in serum free-Dulbecco's modified Eagle's medium in the presence or absence of KME (10, 30, 100, and 300 μg/mL), then further treated with PDGF-BB (10 ng/mL). VSMC proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and VSMC migration was determined using the Boyden chamber and scratch wound healing assays. Western blot analysis was used to detect phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2), protein kinase B (p-Akt), and stress-activated protein kinase/c-Jun NH2-terminal kinase (p-SAPK/JNK). The antimigration and proliferation effects of KME were tested using aortic sprout outgrowth.@*RESULTS@#The HPLC analysis identified honokiol (0.45 mg/g) and magnolol (0.34 mg/g) as the major components of KME. KME (30, 100, and 300 μg/mL) significantly decreased the proliferation and migration of PDGF-BB-stimulated (10 ng/mL) VSMCs and the PDGF-BB-induced phosphorylation of EKR1/2, Akt, and SAPK/JNK (P<0.05). Furthermore, PDGF-BBinduced VSMCs treated with 300 μg/mL of KME showed reduction in aortic sprout outgrowth.@*CONCLUSION@#KME could inhibit abnormal proliferation and migration of VSMCs by down-regulating the phosphorylation of EKR1/2 and Akt. Thus, KME might be a functional food for preventing vascular disorders.

Berberine Inhibits Mechanical Stretch Stress induced MAPK Phosphorylation，Proliferation and Migration of Vascular Smooth Muscle Cells / 中山大学学报(医学科学版)

@#【Objective】To observe whether berberine can inhibit vascular smooth muscle cells（VSMC）proliferation induced by mechanical strength stress and to investigate the role of MAPK pathway in it.【Methods】The cultured VSMC were divided into 4 groups：negative control group（NC group），stretch stress group（SS group），berberine pretreated and stretch stress stimulation group（BBR+SS group），and berberine group. In NC group，phosphate buffer saline was used as a negative control；in SS group，stretch stress was given to VSMC；in BBR+SS group，VSMC were pretreated with berberine for 1 hour and then exposed to stretch stress；in BBR group，VSMC were treated only with berberine for 1 hour and cultured in serum- free DMEM afterwards. We collected VSMC in each group ，detected and analyzed their MAPK phosphorylation，proliferation and migration by using Western blotting，immunofluorescence and wound-healing assay respectively. 【Results】 Compare with NC group，stretch stress markedly induced VSMC proliferation and migration ，which could be inhibited significantly by berberine. Stretch stress obviously increased phosphorylation of MAPK （ERK，JNK，p38），which could be inhibited by berberine in a concentration dependent manner. 【Conclusion】 Berberine inhibits hypertension-induced proliferation and migration of VSMC through MAPK pathway. The results revealed the new use and mechanism of berberine，and provided important data for further study on the prevention and treatment of vascular remodeling caused by abnormal increase of mechanical stress in hypertension.

The effect of PKD1 gene on autophagy in aortic smooth muscle cells / 中国胸心血管外科临床杂志

@#Objective    To explore the effects of PKD1 gene on mouse aortic smooth muscle (MOVAS) cells autophagy. Methods    The shRNA and over-expression lentiviral vectors for the target gene of PKD1 were constructed. MOVAS cells were infected by a number of successful packaging shRNA (PKD1 knockdown) or ETS-1 (PKD1 over-expressing) lentiviral vectors, and qPCR was used to test interference and over-expressing effects. Then qPCR and Western blotting were used to detect the expression levels of autophagy markers including Atg5, Beclin1 and LC3 in control group, shPKD1 group and ETS-1 group. Results    Compared with the control group, PKD1 mRNA level was decreased in the shPKD1 group (P<0.05); ETS-1 and PKD1 mRNA levels were increased in the ETS-1 group (P<0.05). In contrast with the control group, the mRNA levels of autophagy markers including Atg5 (P<0.05) and Beclin1 (P<0.01) were obviously decreased in the shPKD1 group, but they were obviously increased in the ETS-1 group (P<0.001). Protein levels of Atg5, Beclin1 and LC3 were significantly decreased in the shPKD1 group (P<0.05), but they were increased obviously in the ETS-1 group (P<0.05) in contrast with the control group. Conclusion    PKD1 gene is involved in MOVAS cells autophagy, low expression of PKD1 gene can inhibit autophagy and high expression of PKD1 promotes autophagy in vascular smooth muscle cells.

Inhibition Effect of Ginkgo biloba Extract(EGb761) on the Phenotypic Switch of Vascular Smooth Muscle Cells through Regulation of APMK/KLF4 Signaling Pathway / 中国药学杂志

OBJECTIVE: To explore the effects of Ginkgo biloba extract (EGb761) on platelet derived growth factor (PDGF)-induced phenotypic switch of vascular smooth muscle cells (VSMCs) and its potential mechanisms. METHODS: VSMCs were cultured in vitro, and 20 ng•mL-1 PDGF was used to induce the phenotypic switch of VSMCs. MTT assay and wound healing assay were performed to determine the effects of various concentration (1, 10, 100 μg•mL-1) of EGb761 on cell proliferation and migration, respectively; immunofluorescence and Western blot assay were used to detect the arrangement of myofilament, the expression of phenotypic proteins including α-SMA, calopnin and OPN, as well as the protein expression of AMPK/KLF4 signaling pathway. RESULTS: Compared with the control group, PDGF significantly promoted the proliferation and migration of VSMCs. However, EGb761 treatment inhibited PDGF-induced cell proliferation and migration in a concentration-dependent manner. Compared with the control group, PDGF treatment induced disordered arrangement of myofilament and reduced the fluorescence intensity of F-actin. In addition, PDGF significantly decreased the expression of α-SMA and calponin, whrease increased the expression of OPN in VSMCs, when compared with the control group. VSMCs in PDGF+EGb761 group showed well-aligned myofilament and enhanced the fluorescence intensity of F-actin; the expressions of α-SMA and calponin were increased and OPN was decreased in the PDGF+EGb761 group when compared with the PDGF group. Meanwhile, as compared with the control group, PDGF increased the level of phosphorylated AMPK and the expression levels of KLF4, which was inhibited by the addition of EGb761 in a concentration dependent manner. After inhibition of AMPK/KLF4 signaling pathway with the use of specific AMPK pathway inhibitor compound C, the inhibitory effect of EGb761 on PDGF-mediated phenotypic switch of VSMCs was enhanced; vice versa, the activation of AMPK/KLF4 signaling pathway with AMPK pathway activator AICAR and the inhibitory effect of EGb761 on PDGF-mediated phenotypic switch of VSMCs were reversed. CONCLUSION: EGb761 inhibits PDGF-mediated phenotypic switch of VSMCs by targeting APMK/KLF4 signaling pathway.

Influence of influenza A and B virus infections on human aortic smooth muscle cells and the expressions of cytokines / 医学研究生学报

Objective　Vascular smooth muscle cells are the main cells in atherosclerosis. Reports are rarely seen on influenza virus infection on human aortic smooth muscle cells (HASMC) and its influence on the expressions of the related cytokines. This study was to investigate the impact of influenza A virus (IAV) and influenza B virus (IBV) infection on HASMCs and the expressions of cytokines. Methods　HASMCs were stimulated with IAV or IBV or not stimulated with virus (the control). The nucleoprotein of the influenza virus in the cells was detected by immunofluorescence assay, the proliferation of the cells determined with CCK8, and the level of influenza virus RNA in the supernatant measured by qPCR. The collected supernatant was used to infect Madin-Darby canine kidney (MDCK) cells and detect the influenza virus nucleoprotein. The expressions of the cytokines of the influenza virus after 24 hours of infection were determined by qPCR.　Results　At 3 and 4 days after infected with influenza virus, the proliferation of the HASMCs was significantly inhibited in the IAV and IBV groups as compared with the control (P<0.05). The expression of virus RNA in the supernatant of the IBV group at 3 days was 5.75 times as high as that at 2 days (P<0.05), dropped at 4 days but still higher than that at 2 days (P<0.01). Compared with the normal culture medium, the medium with virus growth fluid significantly elevated the RNA level of IAV (0.842±0.148 vs 15.182±1.932, P< 0.01) and IBV (0.962±0.033 vs 4.029±0.681, P<0.01). After infection, the expression of MCP-1 was remarkably up-regulated in the IAV and IBV groups (4.364±0.193 and 3.348±0.507) in comparison with that in the control group (1.001±0.001) (P<0.05), and so were the expressions of IL-6 and TNF-α (P<0.05).　Conclusion　Both IAV and IBV can infect HASMCs and increase the expressions of the cytokines MCP-1, IL-6 , and TNF-α.

Effect of miR-34a on Proliferation of Pulmonary Artery Smooth Muscle Cells in Rats Induced by Hypoxia / 中山大学学报(医学科学版)

@#To observe the effect of miR-34a on the proliferation of pulmonary artery smooth muscle cells in rats induced by hypoxia and explore its possible mechanism.【Methods】Rat pulmonary artery smooth muscle cells were primarily isolated from pulmonary arteriole and cultured. After 3% O2 treatment，the expression of miR- 34a and Notch1 mRNA in rat PASMC were detected by real time PCR. The cell proliferation was detected by EDU after over-expression and inhibition of miR-34a and silencing Notch1 by cell transfection under hypoxia，and the expression of PCNA was detected by real time PCR and western blot method.【Results】We successfully isolated and cultured rat PASMC. And after 3% O2 treatment，the expression of miR-34a in rat PASMC was significantly decreased after 48 h compared with 24 h（P < 0.05）. However，the expression of Notch1 mRNA increased significantly after 48 h compared with 24 h（P < 0.05）. In addition， over-expression of miR-34a and silencing Notch1 significantly inhibited hypoxia-induced cell proliferation ，while inhibition of miR-34a significantly promoted the PASMC proliferation（P < 0.05）.【Conclusion】miR-34a participates in the proliferation of PASMC induced by hypoxia，and it may be through up-regulation of Notch1 to induce cell proliferation.