TAM通过PLGF/Flt-1和TGF-β1通路促进非小细胞肺癌生长和肿瘤血管异生的研究

目的 探讨胎盘生长因子(PLGF)介导非小细胞肺癌(NSCLC)细胞和肿瘤相关巨噬细胞(TAM)之间相互作用的具体机制。方法 采用尾静脉注射法将转染腺病毒(AAV)的NSCLC A549细胞注入10周龄的雄性NOD/SCID小鼠体内,构建小鼠肺癌动物模型。流式细胞技术检测瘤体内各种巨噬细胞(MΦ)亚型,RT-qPCR检测肿瘤细胞和TAM中PLGF、血管内皮生长因子受体(Flt-1)和转化生长因子β1(TGF-β1)表达水平。根据A549细胞和MΦ细胞的培养条件不同,分为单纯A549组,单纯MΦ组,A549+MΦ组,A549+MΦ+sFlt-1组(加入10 μg/L sFlt-1),PLGF+MΦ组(加入100 ng PLGF)和PLGF+MΦ+sFlt-1组(加入100 ng PLGF和10 μg/L sFlt-1)。Transwell、MTT法分别检测不同共培养条件下A549细胞的增殖和迁移能力。根据HUVEC细胞的培养条件,分为对照组、条件培养基(CM)组、CM+SB431542组、TGF-β1组和TGF-β1+SB431542组,HUVEC细胞胶原凝胶测定法检测肿瘤血管异生情况。结果 与RFP-细胞相比,PLGF主要由肿瘤细胞表达(31.72±4.69 vs. 2.31±0.06,P＜0.001),而Flt-1主要由CD163+的巨噬细胞表达(P＜0.001)。A549+MΦ组A549细胞增殖能力显著强于单纯A549组和A549+MΦ+sFlt-1组(OD值:3.62±0.23 vs. 4.53±0.34 vs. 3.71±0.37,P＜0.05)。TAM/M2巨噬细胞中TGF-β1的表达量高于M1巨噬细胞(0.99±0.23 vs. 0.07±0.02,P＜0.05)。CM组和TGF-β1组中HUVEC细胞管状结构的形成最多(P＜0.05),TGF-β1抑制剂SB431542可抑制这一现象。结论 TAM和NSCLC细胞间通过PLGF/Flt-1和TGF-β1信号通路间的交互作用,促进NSCLC的生长和血管异生。     

The association of second trimester biomarkers in amniotic fluid and fetal outcome

Objective: To identify the level of amniotic fluid lactate (AFL), placental growth factor (PLGF), and vascular endothelial growth factor (VEGF) at second trimester amniocentesis, and to compare levels in normal pregnancies with pregnancies ending in a miscarriage, an intrauterine growth restricted fetus (IUGR) or decreased fetal movements.

Study design: A prospective cohort study. Amniotic fluid was consecutively collected at amniocentesis in 106 pregnancies. Fetal wellbeing at delivery was evaluated from medical files and compared with the levels of AFL, VEGF, and PLGF at the time of amniocentesis.

Results: The median level of AFL was 6.9?mmol/l, VEGF 0.088?pg/ml, and PLGF 0.208?pg/ml. The median levels of AFL in pregnancies ended in miscarriage were significantly higher (10.7?mmol/l) compared to those with a live new-born (6.9?mmol/L, p?=?.02). The levels of VEGF (p?=?.2) and PLGF (p?=?.7) were not affected. In pregnancies with an IUGR, the median level of AFL was higher compared to those with normal fetal growth (p?=?.003). No differences VEGF (p?=?.5), but significant lower PLGF were found in IUGR pregnancies (p?=?.03).

Conclusions: Pregnancies ending in a miscarriage or with IUGR had significantly higher median values of AFL but lower values of PLGF in the amniotic fluid at the time of second trimester amniocentesis compared to normal pregnancies.     

Suppression of angiogenic response in local vein wall is associated with reduced thrombus resolution

Introduction

The formation of new vascular channels within and around venous thrombus contributes to its resolution. Neovascularisation arising from the surrounding vein may facilitate this process. Treatment of cancer patients with anti-angiogenic agents can lead to increased incidence of venous thromboembolic events, but the effect of these agents on the processes that govern thrombus resolution are unclear. The aim of this study was to determine the effect of anti-angiogenic treatment with 2-methoxyestradiol (2ME) on (i) angiogenic response in the thrombosed vein and (ii) venous thrombus resolution.

Materials and methods

Venous thrombus was induced in the inferior vena cava (IVC) of 36 adult male BALB/C mice. Thrombosed mice received either the anti-angiogenic agent, 2ME (150 mg/kg/day, i/p), or vehicle control (n = 18/group). In the thrombosed IVC of both groups: hypoxia-inducible factor (HIF) 1α, and its angiogenic targets, vascular endothelial growth factor (VEGF) and placental growth factor (PLGF), were quantified using enzyme-linked immunosorbent assays at days 1 and 10 post-thrombus induction (n = 6/group); and inflammatory cell content, cell proliferation, and vein recanalisation were quantified using immunostaining and image analysis at day 10 (n = 6/group).

Results

In the IVC of mice treated with 2ME compared with control: HIF1α (P < 0.005 and P < 0.02), VEGF (P < 0.005 and P < 0.02), and PLGF levels (P < 0.01 and P < 0.001) were reduced at days 1 and 10 post-thrombus induction respectively, and macrophage content (P < 0.005), neutrophil content (P < 0.01), vein recanalistion (P < 0.05), and thrombus resolution (P < 0.001) were also reduced at day 10.

Conclusions

Anti-angiogenic treatment with 2ME supressed the HIF1-mediated angiogenic drive in local vein wall and attenuated venous thrombus resolution. The potential pro-thrombotic effect of anti-angiogenic agents should be carefully considered when managing venous thromboembolic events in cancer patients.     

HDP患者H2S、PLGF联合Lp-PLA2测定对子痫前期的预测价值

目的分析妊娠期高血压(HDP)患者胎盘生长因子(PLGF)、脂蛋白相关磷脂酶A2(Lp-PLA2)联合内源性硫化氢(H₂S)的测定对子痫前期的预测价值。方法选取2017年9月至2020年10月郑州大学第一附属医院收治的102例患有妊娠期高血压疾病孕妇作为研究组,另选取87例同时间段健康妊娠期孕妇作为对照组。比较两组H₂S、PLGF、Lp-PLA2水平,比较发生以及未发生子痫前期患者的H₂S、PLGF、Lp-PLA2水平;分析影响子痫前期发生的危险因素;采用工作特征曲线(ROC)分析H₂S、PLGF、Lp-PLA2及三者联合检测对子痫前期的预测价值。结果研究组Lp-PLA2水平明显高于对照组,H2S、PLGF水平均低于对照组,差异具有统计学意义(P<0.05)。未发生组H₂S、PLGF水平高于发生组,Lp-PLA2水平低于发生组,差异具有统计学意义(P<0.05)。经多因素Logistic回归模型分析结果得出,妊娠期并发症、H₂S、PLGF、Lp-PLA2为影响子痫前期发生的独立危险因素(P<0.05)。ROC曲线显示,H₂S、PLGF、Lp-PLA2三者联合检测子痫前期的AUC值最大(0.947),灵敏度、特异度最高,分别为92.65%、94.43%(P<0.05)。结论H₂S、PLGF联合Lp-PLA2的测定有助于临床预测子痫前期。