本文選題：多發(fā)性硬化　切入點：實驗性自身免疫性腦脊髓炎　出處：《河北醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:研究亞甲藍(methylene blue,MB)對由髓鞘少突膠質(zhì)細胞糖蛋白(myelin oligodendrocyte glycoprotein,MOG)誘導(dǎo)的實驗性自身免疫性腦脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠發(fā)病的影響,觀察小鼠脾組織中Th17、Treg細胞的比例變化,初步探討MB對EAE小鼠的治療作用以及可能免疫機制。方法:1將24只C57BL/6小鼠隨機分為EAE組、MB組和control組,每組分別8只。以MOG35-55為抗原免疫,輔以百日咳毒素、完全弗氏佐劑及結(jié)核菌素,建立小鼠EAE模型,control組不免疫。免疫當天記為第0天,自免疫第1天開始給藥,MB組小鼠給予MB 20mg/kg每日腹腔注射;control組及EAE組小鼠每日給予等體積生理鹽水腹腔注射。觀察小鼠的發(fā)病時間和每日神經(jīng)功能評分。2將18只C57BL/6小鼠隨機分為EAE組、MB組和control組,每組分別6只。以MOG35-55為抗原免疫,輔以百日咳毒素、完全弗氏佐劑及結(jié)核菌素,建立小鼠EAE模型,control組不免疫。免疫當天記為第0天,自免疫第1天開始給藥,MB組小鼠給予MB 20mg/kg每日腹腔注射;control組及EAE組小鼠每日給予等體積生理鹽水腹腔注射。免疫后25天(發(fā)病高峰期)將小鼠處死,無菌條件下取出小鼠的脾臟。流式細胞學方法測定小鼠脾組織細胞中Th17和Treg細胞的比例。結(jié)果:1各組小鼠發(fā)病情況:EAE組和MB組小鼠發(fā)病率均為100%,發(fā)病高峰期在免疫后25天左右。與EAE組相比,MB組發(fā)病高峰期的神經(jīng)功能評分明顯降低(P0.01);MB組的發(fā)病時間較EAE組晚(P0.05)。control組小鼠不發(fā)病;2各組小鼠脾組織中Treg細胞的比例變化:在發(fā)病高峰期,各組小鼠脾組織中Treg細胞比例均有明顯差別。與control組相比,EAE組小鼠脾組織中Treg細胞比例明顯降低(P0.01);與EAE組和control組相比,MB組小鼠脾組織中Treg細胞比例明顯升高(P0.01);3各組小鼠脾組織中Th17細胞的比例變化:在發(fā)病高峰期,與control組相比,EAE小鼠脾組織中Th17細胞比例明顯升高(P0.05);與EAE組對比,MB組小鼠脾組織中Th17細胞比例明顯降低(P0.05);但MB組與control組相比,小鼠脾組織中Th17細胞比例無明顯差異(P0.05)。結(jié)論:1 MB干預(yù)可以降低EAE小鼠的神經(jīng)功能缺損程度,使EAE小鼠的發(fā)病時間推遲。2 MB干預(yù)可能通過影響小鼠脾組織內(nèi)Th17/Treg細胞比例從而使EAE小鼠神經(jīng)缺損程度減輕,推遲發(fā)病時間,但其具體的機制仍待進一步研究。
[Abstract]:Aim: to study the effect of methylene blue (MBB) on the pathogenesis of experimental autoimmune encephalomyelitis induced by myelin oligodendrocyte glycoprotein (MOG) in mice, and to observe the percentage of Th17T Treg cells in spleen tissue of mice. Methods 24 C57BL / 6 mice were randomly divided into EAE group (MB group) and control group (8 mice in each group). The mice were immunized with MOG35-55 as antigen and with pertussis toxin. Complete Freund's adjuvant and tuberculin were used to establish EAE model in mice. The control group was not immunized. The day of immunization was recorded as day 0. Since the first day of immunization, mice in the MB-treated group were given intraperitoneal injection of MB 20mg / kg daily intraperitoneal injection of normal saline in the control group and the EAE group. The onset time and daily neurological function score of 18 C57BL / 6 mice were observed. Mice were randomly divided into EAE group and control group. Six mice in each group were immunized with MOG35-55 as antigen, supplemented with pertussis toxin, complete Freund's adjuvant and tuberculin. Since the first day of immunization, mice in MB-treated group were given intraperitoneal injection of MB 20mg / kg daily intraperitoneal injection of control group and EAE group, and mice in EAE group were given intraperitoneal injection of the same volume of normal saline daily. The mice were killed 25 days after immunization (peak period of onset). The percentage of Th17 and Treg cells in spleen tissue cells of mice was determined by flow cytometry. Results the incidence of Th17 and Treg cells in mice in group 1 was 100, and the peak period of disease was in immunity. Compared with the EAE group, the neurological function score of the peak period of the disease in the MB group was significantly lower than that in the EAE group. The percentage of Treg cells in the spleen tissue of the control group was significantly lower than that of the EAE group, and the percentage of Treg cells in the spleen tissue of the mice in the control group was lower than that in the control group. Compared with the control group, the percentage of Treg cells in the spleen tissue of the control group was significantly lower than that of the EAE group and the control group, and the percentage of Treg cells in the spleen tissue of the MB-MB group was significantly higher than that of the EAE group and the control group. The percentage change of Th17 cells in spleen tissue of mice in group A: during the peak period of onset, Compared with control group, the proportion of Th17 cells in spleen tissue of control mice was significantly higher than that of EAE group, and the percentage of Th17 cells in spleen tissue of EAE group was significantly lower than that of EAE group, but the ratio of Th17 cells in MB group was significantly lower than that in control group. There was no significant difference in the proportion of Th17 cells in spleen tissue of mice (P 0.05). Conclusion the intervention of 1: 1 MB can reduce the degree of nerve function defect in EAE mice. Delaying the onset time of EAE mice with the intervention of 2.2MB may reduce the degree of nerve defect and delay the onset time of EAE mice by affecting the proportion of Th17/Treg cells in the spleen tissue of EAE mice, but the specific mechanism remains to be further studied.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R744.51

【相似文獻】

相關(guān)期刊論文 前10條

1 ;反復(fù)性原發(fā)性血小板減少性紫癜病人中的肝內(nèi)脾組織[J];國外醫(yī)學.外科學分冊;1998年06期

2 邱勇鋼;徐宏偉;;肝內(nèi)異位脾組織1例[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2007年06期

3 黃U，

本文編號：1581628