發(fā)布時(shí)間：2018-11-07 09:09

【摘要】：目的:課題通過(guò)建立淋巴瘤Balb/C小鼠模型,觀察荷瘤模型鼠外周血炎癥因子、凝血指標(biāo)、血小板活化指標(biāo)和免疫指標(biāo)的水平變化,及瘤體組織中腫瘤相關(guān)巨噬細(xì)胞的表達(dá),研究腫瘤免疫炎性微環(huán)境與高凝狀態(tài)的關(guān)系,并初步探討中藥南蛇藤素干預(yù)淋巴瘤的治療機(jī)制。方法:1、本課題選用具有正常免疫功能的Balb/C小鼠,建立B細(xì)胞淋巴瘤荷瘤模型。將25只6-8周齡的Balb/C小鼠隨機(jī)分為對(duì)照組(n=5)、荷瘤干預(yù)組(n=10)及荷瘤未干預(yù)組(n=10)。荷瘤組皮下注射濃度為2×106/0.2 mL的38B9細(xì)胞,對(duì)照組予0.2 mL PBS緩沖液皮下注射;荷瘤第14 d時(shí)進(jìn)行MRI和B超等影像學(xué)檢查,確認(rèn)小鼠腫瘤形成;荷瘤干預(yù)組予以南蛇藤素灌胃干預(yù)10d,觀察小鼠若處在瀕死狀態(tài),及時(shí)處死小鼠,取腫瘤組織行HE及免疫組化染色。2、分別于荷瘤1 d、14 d、21 d經(jīng)小鼠眼球內(nèi)眥靜脈取血,采用ELISA法檢測(cè)血漿中CRP、TNF-α、IL-6、IL-10、TF、vWF等水平。3、采用流式細(xì)胞術(shù)檢測(cè)小鼠外周血調(diào)節(jié)性T細(xì)胞及B細(xì)胞數(shù)量以及P選擇素、GPⅡbⅢa等血小板活化指標(biāo)的表達(dá)水平。4、采用免疫組化法檢測(cè)CSF-1R和CD68在荷瘤小鼠瘤組織中的表達(dá)。結(jié)果:1、荷瘤后14 d Balb/C小鼠皮下瘤塊形成,成瘤率100%,病理特征為淋巴瘤細(xì)胞彌漫分布,且瘤塊大量表達(dá)CD19。2、采用38B9淋巴瘤細(xì)胞成功荷瘤Balb/C小鼠,與荷瘤前相比,荷瘤組小鼠外周血CRP、TNF-α、IL-6水平均上升(P0.05),而IL-10水平降低(P0.05);荷瘤組小鼠血漿TF、vWF水平上升(P0.05);淋巴瘤荷瘤小鼠CRP、TNF-α、IL-6分別與TF、vWF水平呈正相關(guān),IL-10與TF、vWF水平呈負(fù)相關(guān)。3、南蛇藤素干預(yù)后,小鼠腫瘤生長(zhǎng)緩慢,平均生存期明顯延長(zhǎng)(P0.05),且瘤重和瘤體積明顯小于未干預(yù)組(P0.05);干預(yù)組小鼠外周血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);血漿TF水平下降(P0.05),vWF有下降趨勢(shì);CD41/CD62p明顯降低(P0.05),而CD41/CD61無(wú)明顯變化;干預(yù)組小鼠瘤組織CSF-1R、CD68表達(dá)陽(yáng)性率明顯下降(P0.05)。結(jié)論:1、Balb/C小鼠B細(xì)胞淋巴瘤動(dòng)物模型成功建立。2、淋巴瘤中炎性因子與腫瘤高凝狀態(tài)密切相關(guān)。3、南蛇藤素可通過(guò)降低炎性因子分泌、改善免疫炎性微環(huán)境、控制高凝狀態(tài)等途徑有效干預(yù)腫瘤進(jìn)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- 偽 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，

本文編號(hào)：2315876