不同工艺的阿胶对血虚证小鼠的血细胞成分及相关细胞因子影响的比较研究

目的本研究旨在评价高压蒸汽提取工艺阿胶与传统常压提取工艺阿胶产品的补血作用,探究不同工艺阿胶补血药理机制。方法将70只C57BL/6雌性小鼠随机分为正常对照组(Normal)、模型组(Model)、阿胶常压工艺低剂量组(aL)、阿胶常压工艺高剂量组(aH)、阿胶高压工艺低剂量组(AL)、阿胶高压工艺高剂量组(AH)、阳性对照药维生素B12组(VB12),每组10只。除正常组外,其余各组均采用乙酰苯肼(Acetylphenylhydrazine,APH)联合环磷酰胺(Cyclophosphamide,CTX)构建血虚证小鼠模型,模型构建成功后,药物组给予相应药物连续灌胃14天,每天1次。治疗结束后,收集小鼠全血、肾脏及股骨组织。血液分析仪测定小鼠外周血象;ELISA法测定小鼠血清造血功能相关细胞因子水平;Western blot法测定小鼠血清及肾脏组织造血功能相关蛋白含量;HE染色观察骨髓组织病理学改变;免疫组化观察骨髓组织粒细胞-巨噬细胞集落刺激因子(Granulocyte-macrophage colony-stimulating factor,GM-CSF)及粒细胞集落刺激因子(Granulocyte colony-stimulating factor,G-CSF)蛋白表达水平。结果不同工艺的阿胶均可显著升高血虚证小鼠外周血红细胞、白细胞、血红蛋白及淋巴细胞含量,降低外周血中红细胞平均体积和平均血红蛋白含量等血液指标,显著改善血虚证小鼠造血细胞减少、脂肪组织增加等症状,增加小鼠骨髓有核细胞数量,提高骨髓G-CSF及GM-CSF的蛋白含量,增加血清促红细胞生成素(Erythropoietin,EPO)、白细胞介素-3(Interleukin-3,IL-3)及GM-CSF含量,增加肾脏EPO蛋白表达,降低血清肿瘤坏死因子(Tumor necrosis factor-α,TNF-α)和白细胞介素-6(Interleukin-6,IL-6)含量。与常压提取工艺生产的阿胶相比,高压蒸汽提取工艺生产的阿胶在提升血虚证小鼠外周血中血细胞含量,增加骨髓有核细胞数量,提高骨髓G-CSF和GM-CSF的蛋白含量,提升血清中EPO和G-CSF含量,降低IL-6含量等方面的效果更显著。结论不同工艺的阿胶均可明显升高血虚证小鼠血细胞数目及血红蛋白水平,高压蒸汽提取工艺的阿胶补血作用强于传统常压提取工艺的阿胶,其机制与上调小鼠血清中EPO、IL-3、GM-CSF、G-CSF等正向调控造血因子,提高肾脏EPO含量,增加骨髓GM-CSF、G-CSF的蛋白表达,降低TNF-α、IL-6等负向调控造血因子有关。

Three previously undescribed steroidal glycoalkaloids from Solanum lyratum and their anti-tumor activities

In the present study,three previously undescribed steroidal glycoalkaloids(compounds 1–3)were isolated from Solanum lyratum.Their structures were elucidated based on comprehensive spectroscopic data.Their anti-angiogenesis and anti-metastatic activities were evaluated by MTT and wound-healing assays,respectively.Tumor-derived vascular endothelial cells(TdECs),obtained by co-culture of A549 and human umbilical vein endothelial cells(HUVECs),were treated with compounds 1–3.Results showed that compounds 1–3 significantly inhibited the migration of TdECs at 25μM despite the weak cytotoxic activities,which indicated that the compounds exerted anti-tumor activities by inhibiting metastasis,rather than directly inhibiting the proliferation of TdECs.