雷公藤甲素通过抑制破骨细胞生成预防骨丢失的研究

目的 探讨雷公藤甲素抗骨质疏松的作用及机制。方法 建立大鼠老年性骨质疏松模型。40只22月龄雄性SD大鼠随机分为雷公藤甲素（每天15μg/kg腹腔注射）治疗组和生理盐水对照组（每天15μg/kg腹腔注射）,连续8周。采用显微CT分析胫骨近端骨松质的骨密度(BMD)和骨显微结构。WB检测成骨相关蛋白表达水平。TRACP-5b染色法测定破骨细胞数,同时检测骨吸收标志物表达水平。结果 显微CT结果显示,雷公藤甲素治疗组大鼠骨密度、骨体积/总体积比值(Bv/Tv)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)、骨小梁间距(Tb.Sp)均显著高于对照组（P<0.05）。两组的成骨相关蛋白表达水平无明显差异,TRACP-5b染色显示雷公藤甲素减少了体内破骨细胞的数量（P<0.05）,同时血液骨吸收标志物水平也明显降低（P<0.05）。结论 雷公藤甲素通过抑制破骨细胞生成进而对老年性骨质疏松有保护作用。雷公藤甲素可能是治疗老年性骨质疏松症的一种可行方案。     

肿瘤坏死因子相关凋亡诱导配体参与雷帕霉素诱导血管内皮细胞功能损伤的体外研究

目的: 探讨雷帕霉素对内皮细胞凋亡和增殖、迁移能力的影响,以及肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达水平的变化。方法: 用浓度为0、1、10、100 μg/L 的雷帕霉素孵育内皮细胞24 h,应用CCK8法检测血管内皮细胞的增殖能力,Transwell小室和划痕试验检测细胞迁移能力,DAPI染色观察凋亡细胞核形态改变,Western blotting法检测caspase-3活性以显示血管内皮细胞的凋亡,并用 Western blotting检测TRAIL在凋亡的内皮细胞中的表达。结果: 雷帕霉素(1-100 μg/L)能诱导血管内皮细胞凋亡并抑制其迁移能力(P<0.01)。除雷帕霉素1 μg/L外, 10 μg/L和100 μg/L雷帕霉素均能抑制内皮细胞增殖能力(P<0.01),同时雷帕霉素(10-100 μg/L)使TRAIL蛋白表达增加,两者作用均呈浓度依赖性(P<0.01)。结论: 雷帕霉素能诱导内皮细胞发生凋亡并抑制其增殖和迁移能力。TRAIL表达上调与雷帕霉素诱导血管内皮细胞损伤有一定的相关性。     

冬凌草甲素对乳腺癌Bcap37细胞DJ-1蛋白表达的影响

目的探究冬凌草甲素对乳腺癌Bcap37细胞DJ-1蛋白表达的影响及作用机制分析。方法培养乳腺癌Bcap37细胞,设置6个组别:正常对照组、Bcap37-3μmol/L冬凌草甲素、Bcap37-30μmol/L冬凌草甲素、Bcap37-300μmol/L冬凌草甲素、Bcap37-PD98059组和Bcap37-冬凌草甲素+PD98059组。MTT法检测细胞的增殖情况;Western blot检测乳腺癌Bcap37细胞中DJ-1和Erk蛋白的表达情况。结果冬凌草甲素能有效抑制乳腺癌Bcap37细胞的增殖,且呈剂量依赖性;进一步研究结果提示,冬凌草甲素和Erk的抑制剂PD98059均能有效抑制DJ-1和p-Erk蛋白的表达,联合应用冬凌草甲素和PD98059的效果明显优于单独应用PD98059,而与单独应用冬凌草甲素的效果差异无统计学意义(P0.05)。结论冬凌草甲素能有效抑制乳腺癌Bcap37细胞的增殖,其机制可能与抑制Erk信号分子进而影响DJ-1蛋白的表达密切相关,但也存在其他的机制有待进一步研究发现。     

血管生成抑制素和反应停对淋巴管内皮细胞生成的影响

目的：探讨血管生成抑制素和反应停对淋巴管内皮细胞增殖和游走的影响及作用机制。方法：淋巴管内皮细胞取自猪的胸导管。采用划线法和MTT法观察血管生成抑制素和反应停对淋巴管内皮细胞增殖和游走的影响。Hoechst染色检测细胞凋亡。结果．划线法和MTT法均显示血管生成抑制素和反应停对淋巴管内皮细胞的增殖和游走有明显的抑制作用。Hoechst染色证实,经血管生成抑制素和反应停处理后的淋巴管内皮细胞,核周围有凋亡小体。结论：血管生成抑制素和反应停对淋巴管内皮细胞的增殖和游走具有明显的抑制作用,且有剂量依赖性。血管生成抑制素和反应停具有促进细胞凋亡的作用。     

雷公藤甲素抑制肝癌BEL-7402细胞增殖的作用机制

目的:探讨雷公藤甲素抑制肝癌BEL-7402细胞增殖的作用机制.方法:磺酰罗丹明B比色法检测细胞增殖抑制率;Hochest 33258荧光染色观察细胞形态变化;流式细胞术检测细胞DNA含量;免疫印迹检测细胞Bcl-2蛋白表达情况.结果:不同浓度的雷公藤甲素对肝癌细胞的增殖抑制作用呈浓度依赖性;荧光染色后观察到雷公藤甲素作用后细胞核染色质致密浓染,呈凋亡形态学改变;流式细胞术检测到细胞出现较高的亚二倍体峰;雷公藤甲素可使细胞Bcl-2蛋白表达水平降低.结论:雷公藤甲素对肝癌细胞的增殖抑制具有药物浓度依赖性,并可通过下调Bcl-2的表达诱导细胞凋亡.     

雷公藤甲素调节小胶质细胞极化反应保护损伤的神经元

背景：将小胶质细胞从M1表型转变为M2表型被认为是治疗神经退行性疾病的一种有希望的策略。多项研究表明,雷公藤甲素可以抑制神经炎症,改善多种神经退行性疾病,但是其机制尚不明确。目的：探讨雷公藤甲素对脂多糖激活小胶质细胞诱导SH-SY5Y细胞损伤的保护作用及其机制。方法：CCK8法检测BV2细胞活性筛选最佳的雷公藤甲素使用浓度;然后将BV2细胞分为3组：对照组,模型组(1μg/mL脂多糖),雷公藤甲素组(1 nmol/L雷公藤甲素+1μg/mL脂多糖),收集上清液,采用Griess法检测一氧化氮水平,ELISA法检测促炎因子白细胞介素6、肿瘤坏死因子α、白细胞介素1β和抗炎因子白细胞介素10水平;免疫荧光染色和Western blot法检测BV2细胞诱导型一氧化氮合酶和精氨酸酶1的表达。收集3组小胶质细胞条件培养液并分别作用于SH-SY5Y细胞：分为对照条件培养液组、模型条件培养液组和雷公藤甲素条件培养液组,TUNEL染色法检测SH-SY5Y细胞凋亡率,免疫荧光染色和Western blot法检测caspase3、Bax和Bcl-2的表达。结果与结论：(1)与对照组比较,模型组BV2细胞上...     

雷公藤甲素对2型糖尿病大鼠足细胞损伤的影响及机制

目的:探讨雷公藤甲素对2型糖尿病(DM)模型大鼠肾脏的足细胞损伤及其机制.方法:高糖高脂喂养加小剂量链脲佐菌素诱导建立2型糖尿病模型,将模型大鼠随机分为糖尿病组(DM组)和雷公藤甲素治疗组[DT组,200 μg/(kg*d)灌胃给药],另设对照组(NC组).8周末检测血压、血糖、肾重、肾肥大指数、24小时尿白蛋白(24 UAL)、Ccr及肾组织电镜改变.免疫组化染色检测肾组织ED-1表达的变化,免疫印迹法检测肾组织中Nephrin、OPN、TGF-β表达的改变.结果:DM大鼠24 UAL明显高于NC组(P＜0.05),雷公藤甲素治疗组大鼠24 UAL明显低于DM组(P＜0.05).DM组大鼠肾小球ED-1阳性细胞数明显高于NC组(P＜0.01),雷公藤甲素治疗后肾组织巨噬细胞浸润明显低于DM组(P＜0.05).DM组肾组织Nephrin表达较NC组显著降低(P＜0.01),雷公藤甲素可明显恢复肾组织Nephrin 表达(P＜0.01).与NC组比较,DM组肾组织OPN、TGF-β蛋白表达上调(P＜0.01),雷公藤甲素可显著抑制肾组织OPN、TGF-β的过度表达(P＜0.01).结论:雷公藤甲素可能通过抑制肾组织巨噬细胞浸润、炎症反应,从而减轻足细胞损伤,达到肾脏保护作用.     

雷公藤甲素对非小细胞肺癌细胞系A549增殖的抑制和凋亡的诱导

目的探讨雷公藤甲素(TP)通过调控趋化因子受体4(CXCR4)基因表达对人非小细胞肺癌(A549)细胞增殖和凋亡的影响。方法实验分为4组:对照组、TP组(100 nm/L TP处理细胞)、CXCR4+TP组(转染质粒及TP处理细胞)和NC+TP组(转染空载质粒及TP处理细胞)。RT-qPCR和Western blot检测CXCR4表达以及转染效果;MTT法检测细胞增殖;AnnexinⅤ-FITC/PI双染色法检测细胞凋亡;Western blot检测增殖及凋亡相关蛋白表达。结果雷公藤甲素能够抑制A549细胞中CXCR4 mRNA和蛋白的表达(P<0. 05)。雷公藤甲素可抑制A549细胞增殖,诱导其凋亡(P<0. 05)。转染pc DNA-CXCR4能够上调CXCR4 mRNA和蛋白的表达(P<0. 05)。上调CXCR4的表达能够部分逆转雷公藤甲素对A549细胞增殖抑制和凋亡诱导的作用(P<0. 05)。结论雷公藤甲素可能通过下调CXCR4的表达抑制A549细胞增殖,诱导细胞凋亡。     

表阿霉素诱导血管通透性改变的机制

目的:观察乳腺癌常规化疗药物表阿霉素(EPI)对血管内皮细胞通透性的影响,探讨其在静脉注射时造成血管渗漏的机制。方法:用0、0.1、1.0、10、100μg/mlEPI处理人脐静脉内皮细胞株HUVEC-CRL-1730,光学显微镜观察细胞形态改变,Transwell小室检测单层内皮细胞的通透性,MTT法检测细胞增殖效率,RT-PCR和Western blotting检测血管扩张刺激磷蛋白(VASP)mRNA和蛋白表达水平。结果:10μg/mlEPI处理内皮细胞24h后,与对照组相比,EPI能显著抑制内皮细胞增殖(P<0.05),细胞收缩、变形,单层内皮细胞通透性增加(P<0.05);同时,VASP mRNA和蛋白表达水平均降低(P<0.05)。结论:EPI静脉注射造成的血管渗漏可能与EPI抑制细胞增殖以及通过降低VASP蛋白表达导致的内皮细胞通透性增加有关。     

LncRNA CCAT1 在子宫内膜癌组织的表达意义以及雷公藤甲素的干预作用

目的:观察雷公藤甲素对子宫内膜癌组织中长链非编码RNA结肠癌相关转录子1(LncRNA-CCAT1)表达的影响,探讨其在子宫内膜癌组织中的表达意义。方法:C57BL/6小鼠接种子宫内膜癌瘤HEC-1A细胞制备荷子宫内膜癌小鼠模型,成瘤后取40只小鼠随机分为模型组、低剂量雷公藤甲素组(25μg/kg)、中剂量雷公藤甲素组(50μg/kg)和高剂量雷公藤甲素组(100μg/kg)。给药15 d后取小鼠肿瘤组织称重,计算抑瘤率;HE染色观察肿瘤组织形态;TUNEL染色观察肿瘤组织细胞凋亡情况;免疫组织化学法检测肿瘤组织caspase-3蛋白表达水平,定量聚合酶链反应(qPCR)检测肿瘤组织LncRNA CCAT1表达水平。结果:模型组肿瘤组织血管丰富,细胞生长密集,排列不规则;低、中、高剂量雷公藤甲素组小鼠肿瘤组织内血管减少,结构破坏,细胞出现不同程度坏死。与模型组比较,低、中、高剂量雷公藤甲素组小鼠肿瘤组织质量均明显减轻(P0.05),抑瘤率均明显升高(P0.05),凋亡细胞数均明显增加(P0.05),肿瘤组织caspase-3蛋白表达水平均明显升高(P0.05),LncRNA CCAT1表达水平均明显降低(P0.05),呈剂量依赖性。结论:雷公藤甲素可通过降低LncRNA-CCAT1表达,下调caspase-3蛋白表达,促进肿瘤细胞凋亡,对荷子宫内膜癌小鼠移植瘤生长起抑制作用。     

Antiangiogenic properties of silver nanoparticles

Angiogenesis is an important phenomenon involved in normal growth and wound healing processes. An imbalance of the growth factors involved in this process, however, causes the acceleration of several diseases including malignant, ocular, and inflammatory diseases. Inhibiting angiogenesis through interfering in its pathway is a promising methodology to hinder the progression of these diseases. The function and mechanism of silver nanoparticles (Ag-NPs) in angiogenesis have not been elucidated to date. PEDF is suggested to be a potent anti-angiogenic agent. In this study, we postulated that Ag-NPs might have the ability to inhibit angiogenesis, the pivotal step in tumor growth, invasiveness, and metastasis. We have demonstrated that Ag-NPs could also inhibit vascular endothelial growth factor (VEGF) induced cell proliferation, migration, and capillary-like tube formation of bovine retinal endothelial cells like PEDF. In addition, Ag-NPs effectively inhibited the formation of new blood microvessels induced by VEGF in the mouse Matrigel plug assay. To understand the underlying mechanism of Ag-NPs on the inhibitory effect of angiogenesis, we showed that Ag-NPs could inhibit the activation of PI3K/Akt. Together, our results indicate that Ag-NPs can act as an anti-angiogenic molecule by targeting the activation of PI3K/Akt signaling pathways.     

Down-modulation of monocyte transendothelial migration and endothelial adhesion molecule expression by fibroblast growth factor: reversal by the anti-angiogenic agent SU6668

Basic and acidic fibroblast growth factor (bFGF and aFGF, respectively) and vascular endothelial growth factor (VEGF) exert angiogenic actions and have a role in wound healing, inflammation, and tumor growth. Monocytes and endothelial cells are involved in these processes, but the effect of FGF and VEGF on monocyte-endothelial cell interactions has not been defined. We observed that monocyte adhesion to resting or cytokine (tumor necrosis factor-alpha or interleukin-1 alpha)-stimulated human umbilical vein endothelial cells (HUVECs) was markedly inhibited (40 to 65%) by culture (1 to 6 days) of HUVECs with aFGF or bFGF. Monocyte transendothelial migration induced by C5a and chemokines (MCP-1, SDF-1 alpha, RANTES, MIP-1 alpha) was also suppressed (by 50 to 75%) on bFGF-stimulated HUVECs. VEGF did not have these effects at the concentrations used (10 to 20 ng/ml), although like bFGF, it promoted HUVEC proliferation. Culture of HUVECs with bFGF and aFGF significantly down-regulated intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin expression on resting or tumor necrosis factor-alpha-stimulated HUVECs, but had no influence on platelet endothelial cell adhesion molecule (PECAM)-1 and VE-cadherin expression. bFGF also inhibited MCP-1 production by HUVECs. The inhibitory effects of bFGF on monocyte transendothelial migration and adhesion molecule expression were reversed by SU6668, an anti-angiogenic agent and bFGF receptor tyrosine kinase inhibitor. Our results suggest that bFGF and aFGF may suppress endothelial-dependent monocyte recruitment and thus have an anti-inflammatory action during angiogenesis in chronic inflammation but inhibit the immunoinflammatory tumor defense mechanism. However, SU6668 is an effective agent to prevent this down-regulatory action of bFGF on monocyte-endothelial cell interactions.     

新型酪氨酸激酶抑制剂XCF-43b体外抗血管新生研究

目的 探究XCF-43b 体外抗血管新生机理.方法 鸡胚尿囊膜(CAM)检测化合物抑制血管新生能力,MTT检测其对HUVEC细胞增殖影响,划痕实验和微管形成实验检测化合物对HUVEC迁移和微管形成的影响,Western印迹检测VEGFR2信号通路相关蛋白表达情况.结果 XCF-43b能够抑制CAM血管新生,抑制HUVEC细胞增殖的IC50为(28.42±7.23) μmol/L,对在VEGF刺激下的HUVEC的IC50值为(9.03±1.28) μmol/L,此外,2.5 μmol/L的XCF-43b能够抑制HUVEC细胞的迁移和微管形成;且能够抑制VEGFR2及其下游信号因子的激活.结论 XCF-43b通过抑制VEGFR2信号通路来抑制血管新生.     

雷公藤内酯醇抑制Raji细胞增殖和迁移的体外实验研究

 目的： 研究雷公藤内酯醇在体外是否具有抗淋巴瘤细胞增殖和迁移的效应。方法： 采用四甲基偶氮唑蓝（MTT）比色法检测雷公藤内酯醇对Raji细胞增殖的作用；采用Annexin V/PI双标流式细胞术检测雷公藤内酯醇对Raji细胞凋亡的影响；采用流式细胞仪检测雷公藤内酯醇对Raji细胞表面CXCR4受体表达的影响；并采用Transwell微孔隔离室迁移实验观察雷公藤内酯醇对Raji细胞体外迁移作用的影响。结果： ①雷公藤内酯醇能以时间和剂量依赖方式抑制Raji细胞的增殖，24 h和36 h的 IC50值分别为43.06 nmol/L和25.08 nmol/L。② 随雷公藤内酯醇药物浓度的增加和作用时间的延长，Raji细胞凋亡率逐渐增高，且呈时效和量效关系。③ Raji细胞经不同浓度雷公藤内酯醇（12.5、25、50 nmol/L）处理后，CXCR4表达率（34.66%±4.34%、23.74%±1.87%、18.10%±1.18%）明显低于对照组（72.47%±7.28%，P＜0.01），此抑制效应呈剂量依赖性；④ Transwell趋化活性分析显示雷公藤内酯醇能抑制rhSDF-1α对Raji细胞的趋化作用，此抑制效应也呈量效关系。结论： 雷公藤内酯醇具有抗淋巴瘤细胞增殖和诱导凋亡的效应，并能阻断Raji细胞的体外迁移，其机制与其对SDF-1/CXCR4生物学轴的抑制效应有关。     

VE-statin/Egfl7 siRNA inhibits angiogenesis in malignant glioma in vitro

This study investigated the role of VE-statin/Egfl7 and its mechanism in angiogenesis in malignant glioma. Transwell culture plates were used to establish an U251-HUVEC co-culture system, which was used to mimic the interaction between malignant glioma and endothelial cells. Lentiviral vectors expressing VE-statin/Egfl7 siRNA were constructed, and U251 cells and HUVECs were transfected to inhibit VE-statin/Egfl7 expression. The proliferation, adherence, migration, and lumen formation of endothelial cells were assayed to investigate the influence of VE-statin/Egfl7 on angiogenesis in malignant glioma in vitro. Data showed that HUVEC growth was temporarily slowed after silencing the VE-statin/Egfl7 gene but rapidly returned to normal. Although endothelial cell migration was not influenced, cell adherence was markedly inhibited. Furthermore, the endothelial cells failed to generate a capillary-like lumen after VE-statin/Egfl7 gene silencing. Therefore, it can be concluded that VE-statin/Egfl7 may regulate the adherence of endothelial cells, thus playing an important role in endothelium-induced lumen formation during angiogenesis in malignant glioma.     

Myricetin Inhibits Breast Tumor Growth and Angiogenesis by Regulating VEGF/VEGFR2 and p38MAPK Signaling Pathways

Tumor angiogenesis is an important cause of tumor growth and metastasis. Myricetin is a flavonoid component used in traditional Chinese medicine that has been demonstrated to have anticancer activity. However, to the best of our knowledge, the effect of myricetin on tumor angiogenesis remains unknown. The present study reports the identification of myricetin as a potential chemopreventive agent by reason of its inhibition of tumor angiogenesis and demonstrates the anticancer effects of myricetin in vivo. Cell Counting Kit-8 assays revealed that myricetin inhibits the proliferation of tumor cells but not that of human umbilical vein endothelial cells (HUVECs), and a transwell assay demonstrated that myricetin could inhibit the migration of HUVECs. A rat aortic ring assay revealed that myricetin could also affect the development of microvessels and the formation of vascular networks. Further, an ELISA showed that myricetin reduced the levels of vascular endothelial growth factor (VEGF) in vivo and in vitro. Western blot analysis indicated that myricetin could downregulate VEGFR2 and p38MAPK. Therefore, myricetin could significantly inhibit tumor angiogenesis and has potential as a chemopreventive agent because of its inhibition to angiogenesis. Anat Rec, 302:2186–2192, 2019. © 2019 American Association for Anatomy     

An involvement of SR-B1 mediated p38 MAPK signaling pathway in serum amyloid A-induced angiogenesis in rheumatoid arthritis

Serum amyloid A (SAA) has been reported high expression in autoimmune diseases, such as rheumatoid arthritis (RA). However, detailed molecular mechanisms induced by SAA in the pathogenesis of RA are still unclear. Herein, we focused on the role of SAA–SR-B1 mediated p38 MAPK signaling pathway in the process of RA angiogenesis. Our results showed that both SAA and SR-B1 predominantly localized to vascular endothelial cells, lining and sublining layers in RA synovium. In a series of in vitro experiments with human umbilical vein endothelial cells (HUVECs), SAA induced the endothelial cells (ECs) proliferation, migration and tube formation. However, blockage of SR-B1 and p38 MAPK inhibited SAA-induced cells proliferation, migration and tube formation. In conclusion, our data showed a possible molecular mechanism for SAA–SR-B1 induced angiogenesis events via p38 MAPK signaling pathway.     

Effects of Fumonisin B1 on Biomechanics and Cytoskeleton of Human Umbilical Vein Endothelial Cells

Objective Fumonisin B1(FB1)is an important mycotoxin in nature worldwide.The biomechanical properties of cells are closely related to their structure and function,and the cytoskeleton is the structural and functional basis of cells motility,and therefore,from a biomechanical point of view,the purpose of this study is to investigate the effects of FB1 on the biomechanical properties,migration capacity and cytoskeletal structure of human umbilical vein endothelial cells(HUVECs),which may lay an experimental foundation for further exploration of the toxicity mechanism of fumonisin.Methods HUVECs were cultured and treated with different concentrations of FB1.Then,CCK-8 kit was used to detect the effect of FB1 on the survival rate.The osmotic fragility of the cells was measured after treatment with different osmotic pressures for30 min.The cell membrane fluidity was measured by fluorescence polarization method.The cell electrophoretic mobility was measured by cell electrophoretic apparatus.The migration capacity of the cells was observed by scratch repair assay.The changes of reactive oxygen species and cytoskeletal structure were observed by confocal laser scanning microscopy.Finally,the mRNA and protein relative expression levels of cytoskeletal binding proteins were detected by real-time PCR,Western blotting and confocal laser scanning.Results The results of CCK-8 showed that FB1 could significantly inhibit the proliferation of HUVECs in a dose-and time-dependent manner.After treatment of HUVECs with FB1,the hypotonic resistance of the cell,cell surface charge,cell membrane fluidity and migration capacity were all weakened,while reactive oxygen species were significantly increased and the cytoskeletal structure was significantly reorganized.Furthermore,RTPCR results showed that the mRNA relative expression levels of cytoskeletal binding proteins,exception of actin,were down-regulated after treated with FB1.Besides,Western blotting and statistical analysis based on fluorescence intensity of laser confocal microscopy confirmed theses changes in protein level.Conclusions FB1 can significantly affect the biomechanical properties and motility of HUVECs,which may be directly correlated to the remodel of F-actin cytoskeleton,as well as the relative expression changes of cytoskeletal binding proteins.It is significant for further exploring the toxicity mechanism of fumonisin.     

Aliskiren抑制LPS诱导HUVECs新生血管的形成及可能机制

 目的: 研究阿利吉仑(aliskiren)对脂多糖(LPS)诱导人脐静脉内皮细胞(HUVECs)新生血管形成能力的影响及可能的机制。方法: 常规培养的HUVECs随机分为空白组和肾素组,ELISA法测定炎性细胞因子肿瘤坏死因子-α(TNF-α)和细胞间黏附分子-1(ICAM-1)水平,Western blot法检测Toll样受体4(TLR4)和ICAM-1的蛋白水平。将常规培养的HUVECs随机分为空白对照组、LPS模型组以及aliskiren低剂量(1μ mol/L)、中剂量(10μ mol/L)和高剂量(100μ mol/L)组。MTT法和BrdU法检测HUVECs的增殖能力,Transwell法测定HUVECs的迁移率,以HUVECs在Matrigel胶上形成管腔结构情况来判断其血管形成能力。ELISA测定炎性细胞因子TNF-α、ICAM-1和单核细胞趋化蛋白-1(MCP-1)的水平,RT-PCR和Western blot法检测肾素、TLR4、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)的mRNA和蛋白水平。结果: 肾素能够刺激HUVECs炎症因子的分泌及TLR4的表达;aliskiren呈浓度依赖性抑制HUVECs增殖、迁移及新生血管形成,降低MCP-1、TNF-α、IL-6水平及肾素、MMP-2、MMP-9的表达,抑制TLR4表达(P<0.05)。结论: Aliskiren能够有效抑制LPS诱导HUVECs新生血管形成能力,可能与其下调肾素表达抑制TLR4途径介导的炎症反应及MMP-2、MMP-9生成有关。     

Role of midkine-progranulin interaction during angiogenesis of hepatocellular carcinoma

Midkine (MK) is a heparin-binding growth factor involved in growth, survival, migration, and differentiation of various target cells and dysregulation of MK signaling is implicated in a variety of inflammatory diseases and cancers. Although MK has been reported to act on endothelial cells and to have proangiogenic effects, the exact role of MK in angiogenesis is poorly defined. Progranulin (PGRN) is a secreted glycoprotein that functions as an important regulator of development, cell cycle progression, cell motility, tumorigenesis, angiogenesis. We screened the PGRN from the hepatic cancer cell cDNA library which was interacted with MK, and confirmed the binding by co-immunoprecipitation and co-localization. During our study, the interaction between MK and PGRN had the important role on the HUVECs proliferation, migration, and tubulogenesis, which indicated the interaction may regulate the angiogenesis, also the in vivo angiogenesis model CAM showed the promotion effect stimulated by MK and PGRN. These findings provide the first evidence linking the association of MK and PGRN and may identify the mechanism of MK during the hepatocellular carcinoma angiogenesis.