UNASSIGNED: A subcutaneous graft model of breast cancer was established with 4T1 breast cancer cell line in female BALB/c mice aged 6-8 weeks. The mice were randomly assigned to three groups, a tumor-bearing group (TG), peritumoral electroacupuncture tumor-bearing group (EATG), and bevacizumab tumor-bearing group (BTG), with 18 mice in each group. The TG mice did not receive any intervention, the EATG mice received peritumoral electroacupuncture for 30 minutes, and the BTG mice were intraperitoneally injected with bevacizumab at 10mg/kg. Immunofluorescence was performed to assess the expression of CD31/alpha smooth muscle actin (α-SMA) and hypoxia-inducible factor 1-alpha (HIF-1α) in the tumor tissue at various points of time, including before intervention and 3 days and 5 days after intervention. Then, 3 days after intervention, observation of morphological changes of the microvessels in the tumor tissue was performed through Hematoxylin and Eosin (HE) staining and scanning electron microscope.
UNASSIGNED: There was no significant difference in the expression of CD31, α-SMA, and HIF-1α in the tumor tissues of all groups before experimental intervention ( P>0.05). On day 3 of the experimental interventions, the CD31 and HIF-1α expression levels in the tumor tissues of the EATG and BTG mice were significantly reduced ( P<0.01), while α-SMA expression levels were significantly increased ( P<0.01) in both groups. On day 5 of the experimental interventions, the CD31 and HIF-1α expression levels in the tumor tissues of the EATG and BTG mice were still significantly lower than those in the TG mice ( P<0.01), while the α-SMA expression level was significantly higher than that in the TG group ( P<0.05). On day 3 of the experimental interventions, H&E staining showed visible microvessels in the tumor tissues of all 3 groups. In addition, scanning electron microscopic observation showed that the tumor microvessel walls of the TG mice were rough and defective, and that obvious deformities appeared in the lumen. In contrast, the walls of the microvessels of the EATG and BTG mice were generally intact and there was no obvious deformities in the lumen.
UNASSIGNED: Peritumoral electroacupuncture may induce microvasculature normalization by decreasing microvascular density and increasing pericyte coverage of the neovasculature, thereby improving hypoxic microenvironment of breast cancer in mice.
■在6-8周龄的雌性BALB/c小鼠中,用4T1乳腺癌细胞系建立了乳腺癌的皮下移植模型。将小鼠随机分为三组,荷瘤组(TG),瘤周电针荷瘤组(EATG),贝伐单抗荷瘤组(BTG),每组18只小鼠。TG小鼠没有接受任何干预,EATG小鼠接受瘤周电针30分钟,BTG小鼠腹腔注射10mg/kg的贝伐单抗。免疫荧光法检测CD31/α平滑肌肌动蛋白(α-SMA)和缺氧诱导因子1-α(HIF-1α)在不同时间点肿瘤组织中的表达,包括干预前和干预后3天和5天。然后,干预后3天,通过苏木精和伊红(HE)染色和扫描电子显微镜观察肿瘤组织中微血管的形态变化。
■CD31、α-SMA的表达无显著差异,实验干预前各组肿瘤组织中HIF-1α的含量(P>0.05)。在实验干预的第3天,EATG和BTG小鼠肿瘤组织中CD31和HIF-1α的表达水平显著降低(P<0.01),而α-SMA表达均显著升高(P<0.01)。在实验干预的第5天,EATG和BTG小鼠肿瘤组织中CD31和HIF-1α的表达水平仍显著低于TG小鼠(P<0.01),而α-SMA表达水平明显高于TG组(P<0.05)。在实验干预的第3天,H&E染色显示3组肿瘤组织中可见微血管。此外,扫描电镜观察显示TG小鼠肿瘤微血管壁粗糙,管腔出现明显的畸形。相比之下,EATG和BTG小鼠的微血管壁基本完整,管腔内无明显畸形。
■肿瘤周围电针可以通过降低微血管密度和增加新血管的周细胞覆盖率来诱导微血管正常化,从而改善小鼠乳腺癌的缺氧微环境。