重组人血管内皮抑素对接种泡状棘球蚴沙鼠VEGF、CD34表达的影响

发布时间：2018-04-15 17:22

本文选题：泡状棘球蚴病 + 血管生成抑制剂　；参考：《石河子大学》2017年硕士论文

【摘要】：目的研究重组人血管内皮抑素对接种泡状棘球蚴沙鼠血管内皮生长因子(VEGF)、微血管密度(MVD)-CD34表达的影响。方法将50只健康雌性长爪沙鼠随机分为5组:模型组(A组)、生理盐水对照组(B组)、小剂量恩度注射组(C组)、中等剂量恩度注射组(D组)、大剂量恩度注射组(E组),每组10只。采用开腹肝穿刺法接种磷酸缓冲盐溶液(PBS)处理的泡球蚴原头节混悬液建立肝泡球蚴动物模型,A组造模完成后不予任何处理,C、D、E组在造模30天后分别给与5mg/kg、10mg/kg、15mg/kg的恩度溶液200μL腹腔注射,1次/天,连用14天后停7天,21天为一个周期;B组在造模30天后腹腔注射等量生理盐水,方法同恩度组;用药3个疗程后动脉取血并解剖沙鼠,留取肝泡球蚴组织,称量囊湿重,以酶联免疫吸附法(Elisa)、苏木精—伊红(HE)染色、免疫组化及蛋白印迹(Western Blot)检测各组沙鼠肝泡状棘球蚴组织中VEGF、CD34的表达。结果不同剂量恩度组的泡球蚴组织湿重均下降,中等剂量、大剂量恩度组与模型组、生理盐水对照组之间囊湿重差异有统计学意义(P0.05)。各组沙鼠血清中VEGF及CD34的表达量呈递减趋势:除模型组与生理盐水对照组、中等剂量与大剂量恩度组之间VEGF表达无显著性差异外(P0.05),其他各组之间两两比较差异均有统计学意义(P0.05);血清中CD34表达量:中等剂量恩度组、大剂量恩度组分别与模型组、生理盐水对照组之间两两比较差异有统计学意义(P0.05),其他各组之间两两比较差异无统计学意义(P0.05)。各组沙鼠泡球蚴组织中均检测到VEGF和CD34的表达,阳性细胞呈黄色或棕褐色,主要定位于肝泡球蚴组织增殖浸润区的内皮细胞的细胞浆,也可见于细胞膜;中等剂量、大剂量恩度组VEGF免疫组化评分低于模型组、生理盐水对照组,差异有统计学意义(P0.05),小剂量恩度组与模型组、生理盐水对照组无明显差异(P0.05);泡球蚴组织中的MVD值结果显示:恩度各组较模型组、生理盐水组MVD值均下降,差异有统计学意义(P0.05),恩度各组之间、模型组与生理盐水组之间差异无统计学意义(P0.05)。蛋白印迹试验同样检测到VEGF、CD34蛋白含量有所下降:除模型组与生理盐水对照组、中等剂量与大剂量恩度组之间VEGF表达量差异无统计学意义外(P0.05),其他各组之间均有显著性差异(P0.05);CD34的蛋白表达量显示,中等剂量、大剂量恩度组分别与模型组、生理盐水对照组、小剂量恩度组之间两两比较差异有统计学意义(P0.05),其他各组之间两两比较差异无统计学意义(P0.05)。结论重组人血管内皮抑素能够降低泡状棘球蚴模型沙鼠体内血管内皮生长因子的表达,降低微血管密度,在一定程度上抑制泡状棘球蚴的生长。
[Abstract]:Objective to study the effect of recombinant human vascular endostatin on the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD- CD34) in gerbils inoculated with Echinococcus alveolar Echinococcus.Methods Fifty healthy female gerbils were randomly divided into five groups: model group A, saline control group B, low dose Endrate injection group C group, middle dose Endrate injection group D group, high dose Endrate injection group E group with 10 rats in each group.Animal model of hepatic alveolar echinococcosis was established by inoculating phosphate buffer salt solution (PBS) with open liver puncture method to establish the animal model of hepatic alveolar echinococcosis A group A was treated with 5 mg / kg 10 mg / kg 10 mg / kg / kg of 15 mg / kg en solubility after 30 days of model making without any treatment.Liquid 200 渭 L was injected intraperitoneally once a day,The rats in group B were injected intraperitoneally with the same amount of normal saline 30 days after the model was made, and the arterial blood was taken and dissected after 3 courses of treatment, and the hydatid tissues of hepatic alveolus were retained to weigh the wet weight of the capsule.The expression of VEGF CD34 in rat liver alveolar echinococcus was detected by Elisa and Elisa staining, immunohistochemistry and Western blot analysis.Results the wet weight of hydatid tissues in different dose groups decreased, while in medium dose group, there was significant difference in wet weight between large dose group and model group and normal saline control group (P 0.05).The expression of VEGF and CD34 in serum of gerbils in each group showed a decreasing trend: except model group and normal saline control group,There was no significant difference in the expression of VEGF between the middle dose group and the high dose group (P 0.05), but there was significant difference between the other two groups (P 0.05). The expression of CD34 in serum: medium dose group, high dose group and model group, respectively.There was significant difference between normal saline control group (P 0.05) and other groups (P 0.05).The expression of VEGF and CD34 was detected in the tissues of gerbils in each group. The positive cells were yellow or brown, mainly located in the cytoplasm of endothelial cells in the proliferative and infiltrating areas of hepatic alveolar hydatid tissues, and also in the cell membrane.The VEGF immunohistochemical score in the high dose Endol group was lower than that in the model group and the normal saline control group, the difference was statistically significant (P 0.05), and the low dose Endol group and the model group were significantly lower than those in the control group.The results of MVD in alveolar hydatid tissue showed that the MVD value of en group was lower than that of model group, and the difference was statistically significant (P 0.05).There was no significant difference between model group and saline group (P 0.05).Western blotting test also detected a decrease in VEGF CD34 protein content: except for model group and saline control group,There was no significant difference in the expression of VEGF between the middle dose and the high dose group (P 0.05), but there was significant difference between the other groups. The protein expression of P0.05 + CD34 in the middle dose group and the large dose group were respectively compared with the model group and the normal saline control group.There was significant difference between two groups (P 0.05), but there was no significant difference between other groups (P 0.05).Conclusion Recombinant human vascular endostatin can reduce the expression of vascular endothelial growth factor (VEGF), reduce the microvessel density and inhibit the growth of hydatid alveolar echinococci to some extent.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R532.32

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