淋巴瘤荷瘤小鼠炎症与肿瘤高凝状态相关性及南蛇藤素干预研究
发布时间：2018-11-07 09:09
【摘要】：目的:课题通过建立淋巴瘤Balb/C小鼠模型,观察荷瘤模型鼠外周血炎症因子、凝血指标、血小板活化指标和免疫指标的水平变化,及瘤体组织中肿瘤相关巨噬细胞的表达,研究肿瘤免疫炎性微环境与高凝状态的关系,并初步探讨中药南蛇藤素干预淋巴瘤的治疗机制。方法:1、本课题选用具有正常免疫功能的Balb/C小鼠,建立B细胞淋巴瘤荷瘤模型。将25只6-8周龄的Balb/C小鼠随机分为对照组(n=5)、荷瘤干预组(n=10)及荷瘤未干预组(n=10)。荷瘤组皮下注射浓度为2×106/0.2 mL的38B9细胞,对照组予0.2 mL PBS缓冲液皮下注射;荷瘤第14 d时进行MRI和B超等影像学检查,确认小鼠肿瘤形成;荷瘤干预组予以南蛇藤素灌胃干预10d,观察小鼠若处在濒死状态,及时处死小鼠,取肿瘤组织行HE及免疫组化染色。2、分别于荷瘤1 d、14 d、21 d经小鼠眼球内眦静脉取血,采用ELISA法检测血浆中CRP、TNF-α、IL-6、IL-10、TF、vWF等水平。3、采用流式细胞术检测小鼠外周血调节性T细胞及B细胞数量以及P选择素、GPⅡbⅢa等血小板活化指标的表达水平。4、采用免疫组化法检测CSF-1R和CD68在荷瘤小鼠瘤组织中的表达。结果:1、荷瘤后14 d Balb/C小鼠皮下瘤块形成,成瘤率100%,病理特征为淋巴瘤细胞弥漫分布,且瘤块大量表达CD19。2、采用38B9淋巴瘤细胞成功荷瘤Balb/C小鼠,与荷瘤前相比,荷瘤组小鼠外周血CRP、TNF-α、IL-6水平均上升(P0.05),而IL-10水平降低(P0.05);荷瘤组小鼠血浆TF、vWF水平上升(P0.05);淋巴瘤荷瘤小鼠CRP、TNF-α、IL-6分别与TF、vWF水平呈正相关,IL-10与TF、vWF水平呈负相关。3、南蛇藤素干预后,小鼠肿瘤生长缓慢,平均生存期明显延长(P0.05),且瘤重和瘤体积明显小于未干预组(P0.05);干预组小鼠外周血CRP、TNF-α、IL-6等促炎因子水平下降,而IL-10等抑炎因子水平上升(P0.05);血浆TF水平下降(P0.05),vWF有下降趋势;CD41/CD62p明显降低(P0.05),而CD41/CD61无明显变化;干预组小鼠瘤组织CSF-1R、CD68表达阳性率明显下降(P0.05)。结论:1、Balb/C小鼠B细胞淋巴瘤动物模型成功建立。2、淋巴瘤中炎性因子与肿瘤高凝状态密切相关。3、南蛇藤素可通过降低炎性因子分泌、改善免疫炎性微环境、控制高凝状态等途径有效干预肿瘤进程。
[Abstract]:Objective: to observe the changes of inflammatory factors, coagulation index, platelet activation index and immune index in peripheral blood and the expression of tumor-associated macrophages in tumor-bearing Balb/C mice. To study the relationship between tumor immune and inflammatory microenvironment and hypercoagulability, and to explore the therapeutic mechanism of Chinese traditional medicine Nanteng on lymphoma. Methods: 1. Balb/C mice with normal immune function were selected to establish B cell lymphoma bearing tumor model. Twenty-five Balb/C mice aged 6-8 weeks were randomly divided into control group (n = 5), tumor-bearing intervention group (n = 10) and non-tumor bearing group (n = 10). The tumor bearing group was subcutaneously injected with 2 脳 106 / 0.2 mL 38B9 cells and the control group was subcutaneously injected with 0.2 mL PBS buffer solution. The mice in the tumor-bearing intervention group were treated by oral administration of Nanserexin for 10 days. If the mice were on the verge of death, the mice were killed in time. The tumor tissues were taken for HE and immunohistochemical staining. The blood was collected from the medial canthus vein of mice at 21 days. The levels of CRP,TNF- 伪 and IL-6,IL-10,TF,vWF in plasma were detected by ELISA method. Flow cytometry was used to detect the number of regulatory T cells and B cells in peripheral blood of mice, and the expression levels of P-selectin, GP 鈪，
本文编号：2315876
[Abstract]:Objective: to observe the changes of inflammatory factors, coagulation index, platelet activation index and immune index in peripheral blood and the expression of tumor-associated macrophages in tumor-bearing Balb/C mice. To study the relationship between tumor immune and inflammatory microenvironment and hypercoagulability, and to explore the therapeutic mechanism of Chinese traditional medicine Nanteng on lymphoma. Methods: 1. Balb/C mice with normal immune function were selected to establish B cell lymphoma bearing tumor model. Twenty-five Balb/C mice aged 6-8 weeks were randomly divided into control group (n = 5), tumor-bearing intervention group (n = 10) and non-tumor bearing group (n = 10). The tumor bearing group was subcutaneously injected with 2 脳 106 / 0.2 mL 38B9 cells and the control group was subcutaneously injected with 0.2 mL PBS buffer solution. The mice in the tumor-bearing intervention group were treated by oral administration of Nanserexin for 10 days. If the mice were on the verge of death, the mice were killed in time. The tumor tissues were taken for HE and immunohistochemical staining. The blood was collected from the medial canthus vein of mice at 21 days. The levels of CRP,TNF- 伪 and IL-6,IL-10,TF,vWF in plasma were detected by ELISA method. Flow cytometry was used to detect the number of regulatory T cells and B cells in peripheral blood of mice, and the expression levels of P-selectin, GP 鈪，
本文编号：2315876
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