Inhibitive effects of anti-oxidative vitamins on mannitol-induced apoptosis of vascular endothelial cells

Objective: Study blood vessel injury and gene expression indicating vascular endothelial cell apoptosis induced by mannitol with and without administration of anti-oxidative vitamins. Methods: Healthy rabbits were randomly divided into four groups. Mannitol was injected into the vein of the rabbit ear in each animal. Pre-treatment prior to mannitol injection was per- formed with normal saline (group B), vitamin C (group C) and vitamin E (group D). Blood vessel injury was assessed under electron and light microscopy. In a second experiment, cell culture specimen of human umbilical vein endothelial cells were treated with mannitol. Pre-treatment was done with normal saline (sample B), vitamin C (sample C) and vitamin E (sample D). Total RNA was extracted with the original single step procedure, followed by hybridisation and analysis of gene expression. Results: In the animal experiment, serious blood vessel injury was seen in group A and group B. Group D showed light injury only, and normal tissue without pathological changes was seen in group C. Of all 330 apoptosis-related genes analysed in human cell culture specimen, no significant difference was seen after pre-treatment with normal saline, compared with the gene chip without pre-treatment. On the gene chip pre-treated with vitamin C, 45 apoptosis genes were down-regulated and 34 anti-apoptosis genes were up-regulated. Pre-treatment with vitamin E resulted in the down-regulation of 3 apoptosis genes. Conclusion: Vitamin C can protect vascular endothelial cells from mannitol-induced injury.

Size-dependent Biological Effects on Vascular Endothelial Cells Induced by Different Particulate Matters

Summary： The contribution of particles to cardiovascular mortality and morbidity has been enlightened by epidemiologic and experimental studies. However, adverse biological effects of the particles with different sizes on cardiovascular cells have not been well recognized. In this study, sub-cultured human umbilical vein endothelial cells （HUVECs） were exposed to increasing concentrations of pure quartz particles （DQ） of three sizes （DQPM1, 〈1 μm; DQPM3-5, 3-5 μm; DQPM5, 5 μm） and carbon black particles of two sizes （CB0.1, 〈0.1 μm; CB 1, 〈 1 μm） for 24 h. Cytotoxicity was estimated by measuring the activity of lactate dehydrogenase （LDH） and cell viability. Nitric oxide （NO） generation and cyto- kines （TNF-α and IL-1β） releases were analyzed by using NO assay and enzyme-linked immunoabsorbent assay （ELISA）, respectively. It was found that both particles induced adverse biological effects on HUVECs in a dose-dependent manner. The size of particle directly influenced the biological activity. For quartz, the smaller particles induced stronger cytotoxicity and higher levels of cytokine responses than those particles of big size. For carbon black particles, CB0.1 was more capable of inducing adverse responses on HUVECs than CB 1 only at lower particle concentrations, in contrast to those at higher concentrations. Meanwhile, our data also revealed that quartz particles performed stronger cell damage and produced higher levels of TNF-α than carbon black particles, even if particles size was similar. In conclusion, particle size as well as particle composition should be both considered in assessing vascular endothelial cells injury and inflammation responses induced by particles.

Endothelial progenitor cells:Exploring the pleiotropic effects of statins

Statins have become a cornerstone of risk modification for ischaemic heart disease patients. A number of studies have shown that they are effective and safe. However studies have observed an early benefit in terms of a reduction in recurrent infarct and or death after a myocardial infarction,prior to any significant change in lipid profile. Therefore,pleiotropic mechanisms,other than lowering lipid profile alone,must account for this effect. One such proposed pleiotropic mechanism is the ability of statins to augment both number and function of endothelial progenitor cells. The ability to augment repair and maintenance of a functioning endothelium may have profound beneficial effect on vascular repair and potentially a positive impact on clinical outcomes in patients with cardiovascular disease. The following literature review will discuss issues surrounding endothelial progenitor cell(EPC) identification,role in vascular repair,factors affecting EPC numbers,the role of statins in current medical practice and their effects on EPC number.

Effect of Antioxidants on Endothelial Cell Reactive Oxygen Species (ROD Generation and Adhesion of Leukocytes to Endothelial Cells

Objective To investigatewhether antioxidants inhibit adhesion of leukocytes to endothelium and furthermore, whether all antioxidants regulate NF-KB activation through a redox sensitive mechanism. Methods The effect of the antioxidative substances pyrrolidin dithiocarbamat (PDTC), dichloroisocumarin (DCI), chrysin and probucol on the endothelial leukocyte adhesion were examined under near physiological flow conditions. The antioxidative activity of antioxidants was measured in a DCF fluorescence assay with flow cytometry. The activation of NF-kB in endothelial cells was investigated in a gel shift assay. Results PDTC and probucol did not show an inhibitory effect to the formation of intracellular H2O2 in TNFa activated human vascular endothelial cells (HUVEC) . Chrysin showed a moderate effect. DCI showed a strong antioxidative effect. In contrast, PDTC and chrysin inhibited the adhesion of HL 60 cells to TNFa-stimulated HUVEC. DCI and probucol did not have influence on the adhesion within the area of the examined shear stresses. Only PDTC inhibited the TNFa-induced activation of NF-KB in endothelial cells. Conclusion The inhibition of the endothelial leukocyte adhesion by antioxidative substances is not to be explained by its antioxidative characteristics only. The inhibitory effect of PDTC on NF-KB activation was probably not related to its antioxidative properties. Endothelial cell Antioxidants NF-kappa-B

Cloning and expression of human arresten gene and effect of its recombinant protein on endothelial cell proliferation

To clone human arresten gene and investigate biological activity of the recombinant protein.Methods Human arresten gene was obtained from the plasmid pGEMArr and subcloned into the BamHⅠ and Pst Ⅰ restriction sites of prokaryotic expression vector pRSET containing T7 promoter.The recombinant plasmid pRSETAN was subsequently transformed into the strain E.coli BL21(DE3),and the target gene was expressed under induction of IPTG.The expressed protein was extracted,purified by Ni 2+ chelation affinity chromatography and refoled.The effect of the recombinant protein on proliferation of human umbilical vein endothelial cells (HUVECs) was also analyzed with the MTT assay.Results Endonuclease digesting and DNA sequencing confirmed that the arresten gene was correctly inserted into the expression vector.The recombinant protein was hightly expressed in the form of inclusion body in the host bacteria after induction.SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 26×103 amounted to 27% of the total bacterial proteins.The purity of the expected protein could reach over 96% through affinity chromatography.After renaturation,the recombinant protein could reach over 96% through affinity chromatography.After renaturation,the recombinant protein could significantly suppress proliferation of human umbilical vein endothelia cells(HUVECs) induced by vascular endothelial growth factor(VEGF).Conclusion Human arresten gene was successfully cloned into the expression vector pRSET and expressed at high level in Escherichia coli.Purified and refolded arresten protein could effectively inhibit proliferation of vascular endothelia cells.2 refs.

Efficacy of Bee Products(Anzer Honey,Pollen and Propolis)in Detection and Healing of Damage Induced by Antidiabetic Drug Vildagliptin/Metformin Hydrochloride in Healthy Human Pancreatic Cells:Cytotoxic,Genotoxic and Biochemical Studies

Background and Objective Although drugs are powerful therapeutic agents,they have a range of side effects.These side effects are sometimes cellular and not clinically noticeable.Vildagliptin/metformin hydrochloride is one of the most widely used oral antidiabetic drugs with two active ingredients.In this study,we investigated its harmful effects on the metabolic activation system in healthy human pancreatic cells“hTERT-HPNE”,and we aimed to improve these harmful effects by natural products.To benefit from the healing effect,we used the unique natural products produced by the bees of the Anzer Plateau in the Eastern Black Sea Region of Turkey.Methods Cytotoxic and genotoxic effects of the drug were investigated by different tests,such as MTT,flow cytometry-apoptosis and comet assays.Anzer honey,pollen and propolis were analyzed by gas chromatography/mass spectrometry(G/C-MS).A total of 19 compounds were detected,constituting 99.9%of the samples.Results The decrease in cell viability at all drug concentrations was statistically significant compared to the negative control(P<0.05).A statistically significant decrease was detected in the apoptosis caused by vildagliptin/metformin hydrochloride with the supplementation of Anzer honey,pollen and propolis in hTERT-HPNE cells(P<0.05).Conclusion This study can contribute to other studies testing the healing properties of natural products against the side effects of oral antidiabetics in human cells.In particular,Anzer honey,pollen and propolis can be used as additional foods to maintain cell viability and improve heal damage and can be evaluated against side effects in other drug studies.

Different Responses of Cell Cycle between Rat Vascular Smooth Muscle Cells and Vascular Endothelial Cells to Paclitaxel

Summary： Although previous reports showed dmg-eluting stent （DES） could effectively inhibit neointima formation, in-stent restenosis （ISR） remains an important obstacle. The purpose of this study was to investigate different effects of paclitaxel on proliferation and cell cycle regulators between vascular smooth muscle cells （VSMCs） and vascular endothelial cells （VECs） of rats in vitro. The cultured VSMCs and VECs of rats from the same tissues were examined by using immunohistochemistry, flow cytometry and Western blotting in control and paclitaxel-treated groups. The results showed paclitaxel could effectively inhibit proliferation of VSMCs and VECs. However, as compared with VECs, prolif- eration of VSMCs in paclitaxel-treated group decreased less rapidly. The percentage of cells in G0-G1 and G2-M phases was reduced, and that in S phase increased after treatment for 72 h. The expression of cyclin D1 and B1, p27 and PCNA in VSMCs of paclitaxel-treated group was up-regulated, but that of p21 down-regulated as compared with VECs. It is concluded that there are significant differences in the expression of cell cycle regulators and proliferation rate between paclitaxel-treated VSMCs and paclitaxel-treated VECs, suggesting that the G1 S checkpoint regulated by paclitaxel may play a critical role in the development of complications of DES, which provides new strategies for treatments of ISR.

Cytotoxic effects of docetaxel as a candidate drug of drug-eluting stent on human umbilical vein endothelial cells and the signaling pathway of cell migration inhibition,adhesion delay and shape change

Docetaxel(DTX),a paclitaxel analogue,can efficiently inhibit proliferation of vascular smooth muscle cells and has broadly been used as an antiangiogenesis drug.However,as a candidate drug of drug-eluting stent,the effects of DTX on human umbilical vein endothelial cells(HUVECs)are still not well understood.Herein,we investigated the effects of DTX on proliferation,apoptosis,adhesion,migration and morphology of HUVECs in vitro.We found that DTX had the cytostatic and cytotoxic effects at low and high concentrations,respectively.DTX could inhibit the proliferation and migration of HUVECs,induce HUVECs apoptosis,delay HUVECs adhesion and decrease spreading area and aspect ratio of individual cells.The signaling pathway that DTX led to the migration inhibition,adhesion delay and shape change of HUVECs is the VE-cadherin mediated integrin b1/FAK/ROCK signaling pathway.The study will provide a theoretical basis for the clinical application of DTX.

Boosting of the enhanced permeability and retention effect with nanocapsules improves the therapeutic effects of cetuximab

Objective:The introduction of therapeutic antibodies(tAbs)into clinical practice has revolutionized tumor treatment strategies,but their tumor therapy efficiency is still far below expectations because of the rapid degradation and limited tumor accumulation of tAbs.Methods:We developed a nanocapsule-based delivery system to induce the self-augmentation of the enhanced permeability and retention(EPR)effect.This system constantly penetrated across the blood-tumor barrier into the tumor while avoiding the attack of tAbs by the immune system.The biodistribution and therapeutic effect were tested with single dose administration of nanocapsule-tAbs in vivo.Results:The accumulation of Nano(cetuximab)within subcutaneous PC9 tumors was gradually enhanced over 6 days after single dose administration,which was contrary to the biodistribution of native cetuximab.Nano(cetuximab)accumulated in tumor tissues via the EPR effect and released cetuximab.The released cetuximab acted on vascular endothelial cells to destroy the blood-tumor barrier and induce self-augmentation of the EPR effect,which in turn contributed to further tumor accumulation of long-circulating Nano(cetuximab).Compared with single dose administration of native cetuximab,Nano(cetuximab)showed an effective tumor suppressive effect for 3 weeks.Conclusions:The nanocapsule-based delivery system efficiently delivered tAbs to tum or tissues and released them to boost the EPR effect,which facilitated further tumor accumulation of the tAbs.This novel self-augmentation of the EPR effect facilitated by the biological characteristics of tAbs and nanotechnology contributed to the improvement of the therapeutic effect of tAbs,and stimulated new ideas for antibody-based tumor therapy.

Experimental study on antitumor effect of arsenic trioxide in combination with cisplatin or doxorubicin on hepatocellular carcinoma

INTRODUCTIONThe main component of a traditional Chinese drug 'Pishuang'. arsenic trioxide (As2O3), has obviously selective anti-tumor effect on human hepatocellular carcinoma (HCC)in both in vitro and in vivo studies[1-5]. Due to limited effectiveness when any anti-carcinogen is used alone and obviously increased toxicity when the dose is raised, there is no exception for As2O3. Furthermore, combined chemotherapy contributes to improve therapeutic effectiveness, disperse toxicity and surmount drug-resistance,in which the combination of traditional Chinese and modern medicine has more advantages and characteristics. As a result,we made an experimental study on anti-tumor effect of As2O3in combination with cisplantin (PDD) or doxorubicin (ADM)on HCC. to investigate the possibility of AS2O3 in combination with PDD or ADM and nature of interaction between them,and to provide experimental basis for clinical application.

Effects of aminoguanidine on nitric oxide production induced by inflammatory cytokines and endotoxin in cultured rat hepatocytes

AIM: To study the effects of aminoguanidine (AG) and two L-arginine analogues N(omega)-nitro-L-arginine methyl ester (L-NAME) and N(omega)-nitro-L-arginine (L-NNA) on nitric oxide (NO) production induced by cytokines (TNF-alpha, IL-1 beta, and IFN-gamma) and bacterial lipopolysaccharide (LPS) mixture (CM) in the cultured rat hepatocytes, and examine their mechanisms action. METHODS: Rat hepatocytes were incubated with AG, L-NAME, L-NNA, Actinomycin D (ActD) and dexamethasone in a medium containing CM (LPS plus TNF-alpha, IL-1 beta, and IFN-gamma) for 24h. NO production in the cultured supernatant was measured with the Griess reaction. Intracellular cGMP level was detected with radioimmunoassy. RESULTS: NO production was markedly blocked by AG and L-NAME in a dose-dependent manner under inflammatory stimuli condition triggered by CM in vitro. The rate of the maximum inhibitory effects of L-NAME (38.9%) was less potent than that obtained with AG(53.7%, P 【 0.05). There was no significant difference between the inhibitory effects of AG and two L-arginine analogues on intracellular cGMP accumulation in rat cultured hepatocytes. Non-specific NOS expression inhibitor dexamethasone (DEX)and iNOS mRNA transcriptional inhibitor ActD also significantly inhibited CM-induced NO production. AG(0.1 mmol x L(-1)) and ActD (0.2 ng x L(-1)) were equipotent in decreasing NO production induced by inflammatory stimuli in vitro, and both effects were more potent than that induced by non-selectivity NOS activity inhibitor L-NAME (0.1 mmol x L(-1)) under similar stimuli conditions (P【0.01). CONCLUSION: AG is a potent selective inhibitor of inducible isoform of NOS,and the mechanism of action may be not only competitive inhibition in the substrate level, but also the gene expression level in rat hepatocytes.

Selective and sensitive fluorescence imaging reveals microenvironment-dependent behavior of NO modulators in the endothelial system

Nitric oxide(NO)is a second messenger playing crucial roles in the signaling of a variety of cellular functions.Due to its pathophysiological significance,various NO modulators have been developed to explore NO pathways and some have been used as therapies.These modulators are often used directly to observe pharmacological effects in cell lines,but their actual effect on intracellular NO level is seldom analyzed.Herein,facilitated by a selective and sensitive fluorescence probe,we observed that some NO modulators displayed unexpected behaviors with both NO scavenger carboxy-PTIO and endothelial nitric oxide synthase(eNOS)inhibitor N(u)-nitro-L-arginine methyl ester(L-NAME)failing to decrease intracellular free NO level in EA.hy926 cells while NO donor diethylamine-NONOate(DEA$NONOate)and eNOS activator calcimycin(A23187)failing to increase free NO level in human umbilical vein endothelial cell line(HUV-EC-C),although the reagents were confirmed to work normally in the primary human umbilical vein endothelial cells(primary HUVECs)and RAW 264.7 macrophage cells.Further research suggested that these unusual behaviors might be attributed to the cellular microenvironments including both the NO synthase(NOS)level and the endogenous glutathione(GSH)level.Genetically manipulating eNOS level in both cells restores the expected response,while decreasing GSH level restores the ability of DEA$NONOate to increase NO level in HUV-EC-C.These results reveal that the cellular microenvironment has a profound impact on pharmacological effect.Our study suggests GSH as a reservoir for NO in live cells and highlights the value of chemical probes as valuable tools to reveal microenvironmentdependent pharmacological effects.

Effect of Chinese Herbal Drug-Containing Serum for Activating-Blood and Dispelling-Toxin on ox-LDL-Induced Inflammatory Factors'Expression in Endothelial Cells

Objective： To investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule （芎芍胶囊, XS, for activating-blood） and Huanglian Capsule （黄连胶囊, HL, for dispellingtoxin） on the oxidized low-density lipoprotein （ox-LDL）-induced inflammatory factors in human umbilical vein endothelial cells （HUVECs）. Methods： Thirty-two rats were randomly divided into four groups： the blank control group treated with distilled water, the positive control group treated with simvastatin （1.8 mg/kg）, the test group I treated with Chinese herbal compound of XS （0.135 g/kg）, and the test group 1T treated with Chinese herbal compound of XS （0.135 g/kg） and HL （0.135 g/kg）. All the treatments were administered for 7 successive days by gastrogavage. Rats＇ blood serum was harvested 1 h after the last administration to prepare respective drug- containing serum. HUVECs were exposed to ox-LDL （100 μg/mL） to induce cell injury model and incubated with corresponding drug-containing serum for 24 h. Untreated HUVECs were set for blank control. Levels of interleukin-6 （IL-6）, tumor necrosis factor-α （TNF-α）, and soluble intercellular adhesion molecule-1 （slCAM-1） in supematant of cultured HUVECs were determined by enzyme-linked immunosorbent assay （ELISA）. HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry. Results： Levels of IL-6, TNF-α, and sICAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation （P〈0.01）, while the abnormal elevations, except slCAM-1 in the test group Ⅰ, were all reduced in the treated groups （the positive control and the two test groups） significantly （P〈0.01 or P〈0.05）. Besides, the effect in the test group Ⅱ seemed somewhat higher than that in the test group Ⅰ but with no statistical significance （P〉0.05）. Conclusion： Drug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.

EFFECTS OF HYPOXIC ENDOTHELIAL CELL CONDITIONED MEDIUM ON PROLIFERATION AND COLLAGEN SYNTHESIS OF SMOOTH MUSCLE CELLS AND INHIBITORY EFFECTS OF RADIX SALVIAE MILTIORRHIZAE

The effects of hypoxic endothelial cell conditioned medium (HECCM) on proliferation and collagen synthesis of cultured porcine pulmonary arterial smooth muscle cells (PASMCs) were studied by 3H-thymidine (3H-TdR) and 3H-proline incorporations, image analysis for determination of DNA content and colorimetric assay using MTT, and the inhibitory effects of radix salviae miltiorrhizae (RSM) on them were also investigated. The results showed that HECCM could induce enhancement of the enzymatic activity of mitochondria, increase of the nucleic DNA content and increases of the 3H-TdR and 3H-proline incorporations in PASMCs. The 3H-proline incorporation in PASMCs cultured in HECCM was 1.83 times as much as that cultured in normoxic endothelial cell conditioned medium (NECCM). Compared with the control, Chinese herb medicine RSM could inhibit the proliferation of PASMCs cultured in HECCM and decrease the 3H-prolinc incorporation in PASMCs cultured in both HECCM and NECCM (P< 0.001). However, RSM had no ef fects on the nucleic DNA content and 3H-TdR incorporation into DNA of PASMCs cultured in NECCM. It suggests that hypoxia may stimulate the endothelia to synthesize and secrete some cytokines which can stimulate the proliferation and the synthesis of collagen of PASMCs and RSM can inhibit this process.

奥希替尼在老年非小细胞肺癌患者靶向治疗中的应用效果及对T细胞水平的影响

目的 探讨奥西替尼在老年非小细胞肺癌患者靶向治疗中的效果及对免疫水平的影响。方法 回顾性选择2018年1月至2020年12月老年非小细胞肺癌患者116例研究,根据治疗方法不同分为2组,各58例。对照组采用常规放化疗治疗,观察组在对照组基础上联合奥西替尼治疗,3个月治疗后评估患者效果,比较2组总有效率、T细胞水平(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))、肿瘤标志物水平、不良反应发生率。结果 观察组治疗3个月总有效率为44.8%高于对照组25.9%(P<0.05);2组治疗后3个月CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平均低于治疗前(P<0.05);CD8^(+)水平高于治疗前(P<0.05);观察组治疗后3个月CD3^(+)(58.95±4.21)%、CD4^(+)(32.59±3.11)%、CD4^(+)/CD8^(+)(1.21±0.22)高于对照组(P<0.05);CD8^(+)(26.81±3.32)%低于对照组(P<0.05);观察组干预3个月后CA125(91±8)U/ml、CYFRA21-1(1.26±0.24)μg/L及癌胚抗原(CEA)水平(34±5)μg/L均低于对照组(P<0.05);2组不良反应发生率差异无统计学意义(P>0.05)。结论 奥西替尼用于老年非小细胞肺癌患者靶向治疗中,能获得较好的总有效率,对患者T细胞水平影响较小,可降低肿瘤标志物水平,未增加不良反应发生率,值得临床推广应用。

替雷利珠联合化疗治疗非小细胞肺癌手术患者的效果

目的评价替雷利珠单抗在含铂双药化疗治疗的非小细胞肺癌手术患者中的应用效果。方法选取2022年1月—2023年12月收治的100例拟行手术治疗的非小细胞肺癌患者,根据随机数字表法将其分成两组。对照组50例患者在术前给予含铂双药治疗,观察组50例患者在其治疗基础上加用替雷利珠单抗治疗。比较两组临床疗效、无事件及无疾病生存率、生活质量改善情况、不良反应。结果观察组临床疗效(完全缓解率:20.00%vs.10.00%)及病理评估(主要病理学缓解率:46.00%vs.20.00%)优于对照组(Z=3.484,P<0.001;χ^(2)=7.664,P=0.006);Kaplan-Meier生存分析显示,观察组无事件生存率(84.00%vs.60.00%)及无疾病生存率(78.00%vs.60.00%)均高于对照组(χ^(2)=4.298,P=0.038;χ^(2)=4.783,P=0.029);在生活质量改善率方面,观察组(64.00%)较对照组高(42.00%),差异有统计学意义(χ^(2)=4.857,P=0.028);两组不良反应发生率(18.00%vs.22.00%)比较,差异无统计学意义(χ^(2)=0.250,P=0.617)。结论在含铂双药化疗治疗非小细胞肺癌手术患者中的实施替雷利珠单抗治疗可提高治疗效果,促进生活质量改善,且不会增加不良反应发生风险。

靶向B7-H3抑制人脐静脉血管内皮细胞生长、迁移和血管形成

目的探讨靶向抑制协同信号分子B7-H3对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)生长、迁移及血管生成能力影响。方法使用小干扰RNA(small interfering RNA,siRNA)敲减HUVECs B7-H3分子,采用CCK-8实验检测24 h、48 h和72 h细胞增殖;采用Transwell实验检测24 h细胞迁移;三维细胞培养观察细胞血管生成。结果与空序列转染的阴性对照(siRNA-Control)组比较,siRNA-720、siRNA-1707和siRNA-16903条备选siRNA对B7-H3表达的抑制效果不同,siRNA-1690抑制率明显高于siRNA-720和siRNA-1707,因此,选择使用siRNA-1690序列进行后续实验。CCK-8细胞活力实验结果显示,敲减B7-H3后24 h、48 h和72 h HUVECs增殖能力分别下降24%、22%(P>0.05,与24 h比较)和15%(P<0.05,与48 h比较);Transwell迁移实验表明,与siRNA-Control组对比,敲减B7-H3的HUVECs 24 h迁移能力明显减低(P<0.01);三维细胞培养结果表明,采用si-B7-H3敲减B7-H3基因后,HUVECs血管生成能力明显下降(P<0.01)。结论靶向B7-H3抑制HUVECs生长、迁移及血管生成。

基于人脐静脉内皮细胞的OGD/R模型研究当归-川芎药对中7个活性成分对VEGF-PI3K-AKT/NF-κB信号通路的影响

目的探讨当归-川芎药对中7个活性成分(绿原酸、阿魏酸、咖啡酸、丁烯基苯酞、洋川芎内酯H、洋川芎内酯A、藁本内酯)对VEGF-PI3K-AKT/NF-κB信号通路关键蛋白及其上下游血管活性物质、黏附因子和炎症因子的调控作用。方法构建人脐静脉内皮细胞(HUVEC)的氧糖剥夺/复氧(OGD/R)模型,采用细胞增殖试剂盒(CCK-8法)检测细胞活力,探索7个成分的最佳造模时间;通过检测乳酸脱氢酶(LDH)释放探索最佳给药浓度;采用酶联免疫吸附法(ELISA)检测7个成分对VEGF、VCAM-1、PAI-1、NF-κB、IL-1和IL-6表达的影响;采用逆转录-聚合酶链式反应(RT-PCR)法检测7个成分对VEGF-PI3K-AKT/NF-κB信号通路关键蛋白mRNA表达的影响。结果HUVEC氧糖剥夺6 h再复氧为最佳造模时间。高剂量绿原酸组、阿魏酸组、洋川芎内酯H组,低、中剂量丁烯基苯酞组,中、高剂量洋川芎内酯A、藁本内酯组显著降低LDH漏出率(P<0.05,P<0.01);绿原酸、阿魏酸、洋川芎内酯H部分剂量组细胞中VEGF、ICAM-1、VCAM-1的表达显著降低,绿原酸、阿魏酸、洋川芎内酯H、藁本内酯部分剂量组细胞NF-κB的表达显著下降,绿原酸、咖啡酸、丁烯基苯酞、洋川芎内酯H、洋川芎内酯A部分剂量组细胞中IL-6的表达显著增加,绿原酸、阿魏酸、洋川芎内酯A部分剂量组细胞IL-1表达显著减少,阿魏酸、洋川芎内酯H部分剂量组细胞中PAI-1的表达量显著下降(P<0.05,P<0.01);绿原酸、阿魏酸、咖啡酸、丁烯基苯酞和洋川芎内酯A部分剂量组细胞中ERK、VEGF、NF-κB、VEGFR2和MMP9的mRNA相对表达量显著下调,洋川芎内酯H和洋川芎内酯A部分剂量组细胞中AKT的mRNA相对表达量均显著上调(P<0.05,P<0.01)。结论当归-川芎中的药效成分可能通过抑制黏附因子、炎症因子和VEGF-PI3K-AKT/NF-κB信号通路关键蛋白mRNA的表达,从而发挥抗缺血性中风的作用。

耳垂瘢痕疙瘩的形成机制及临床治疗研究进展

瘢痕疙瘩是机体对真皮损伤的过度组织反应,发生于真皮网状层,表现为超出原始创面边界并侵入周围健康皮肤的隆起性真皮病灶。最突出的特征是成纤维细胞增殖及细胞外基质的广泛沉积。瘢痕疙瘩常见于上胸部、肩部、后背上方以及头颈部,在头颈部时尤其好发于耳朵,会随着时间进行性增大,常伴随疼痛及瘙痒症状,耳垂瘢痕疙瘩有多种治疗方法,但复发率都很高,在临床上如何有效控制瘢痕疙瘩的发生及复发仍然是个难题,针对耳垂瘢痕疙瘩形成机制进行深入研究,有助于确定特定的靶点途径,减少疾病的发生。对比研究耳垂瘢痕疙瘩的各种治疗手段也有助于患者选择最佳治疗方案,减少疾病复发率,提高患者术后生活质量。