Department of Oto-Rhino-Laryngology, West China Hospital, West China Medical School, Sichuan University China, No. 37 Guo Xue Alley, Chengdu City 610041, Sichuan Province, China.
Department of Otorhinolaryngology Head and Neck Surgery, The People's Hospital of Jianyang City, No. 180, Hospital Road, Jianyang City 641400, Sichuan Province, China.
Biosci Rep. 2019 Aug 19;39(8). doi: 10.1042/BSR20191182. Print 2019 Aug 30.
polysaccharide (LBP) has a variety of pharmacological and biological activities such as anti-inflammatory, antioxidation, anti-apoptosis, immune regulation and other pharmacological effects; however, the effect of LBP on infantile hemangioma (IH) was less reported. Primary human hemangioma endothelial cells (HemECs) were isolated from fresh surgical specimens of patients. HemECs was treated with LBP and the changes in proliferative and apoptotic signaling pathways were investigated by performing cell counting kit-8, cloning formation experiment, angiogenesis experiment, flow cytometry, Western blot, immunofluorescence, HE stain and real-time quantitative polymerase chain reaction. We found that LBP potently inhibited the proliferation of HemECs and achieved a low-micromolar IC (45 and 40 μg/ml, the half maximal inhibitory concentration) value and less angiogenesis, however, the IC had no effect on human umbilical vein endothelial cells (HUVECs) viability. LBP treatment induced apoptosis in HemECs, which was supported by positive Annexin-V-FITC staining, the activation of cleaved caspase-3 and Bcl-2-associated X protein (Bax) and the inhibition of B-cell lymphoma/leukemia-2 (Bcl-2). Moreover, the result demonstrated that LBP suppressed the expressions of proliferating cell nuclear antigen (PCNA), Ki67, vascular endothelial growth factor (VEGF), VEGFR2 and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signal pathway. PI3K-specific agonist (IGF-1) had promotive effects on HemECs proliferation, which was reversed by LBP. Our study suggests that the effectiveness of LBP in IHs may be associated with its potent anti-proliferative and apoptotic activities in HemECs. Thus, our findings may provide an effective medicine for IHs treatment.
多糖（LBP）具有多种药理和生物学活性，如抗炎、抗氧化、抗凋亡、免疫调节等药理作用；然而，LBP 对婴幼儿血管瘤（IH）的作用报道较少。从患者的新鲜手术标本中分离出原代人血管瘤内皮细胞（HemECs）。用 LBP 处理 HemECs，通过细胞计数试剂盒-8、克隆形成实验、血管生成实验、流式细胞术、Western blot、免疫荧光、HE 染色和实时定量聚合酶链反应，研究增殖和凋亡信号通路的变化。我们发现 LBP 能有效抑制 HemECs 的增殖，达到低微摩尔 IC（45 和 40 μg/ml，半最大抑制浓度）值和较少的血管生成，但 IC 对人脐静脉内皮细胞（HUVECs）的活力没有影响。LBP 处理诱导 HemECs 凋亡，这得到 Annexin-V-FITC 染色阳性、裂解 caspase-3 和 Bcl-2 相关 X 蛋白（Bax）的激活以及 B 细胞淋巴瘤/白血病-2（Bcl-2）的抑制的支持。此外，结果表明 LBP 抑制了增殖细胞核抗原（PCNA）、Ki67、血管内皮生长因子（VEGF）、VEGFR2 和磷酸肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）信号通路的表达。PI3K 特异性激动剂（IGF-1）对 HemECs 增殖有促进作用，LBP 可逆转其作用。我们的研究表明，LBP 在 IH 中的有效性可能与其在 HemECs 中的强大抗增殖和凋亡活性有关。因此，我们的研究结果可为 IH 的治疗提供一种有效的药物。
