本文選題：抗CD20單克隆抗體　切入點(diǎn)：白介素10　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:觀察抗CD20單克隆抗體聯(lián)合腺病毒介導(dǎo)的IL-10基因?qū)Πl(fā)病早期非肥胖性糖尿病鼠(NOD)肝臟細(xì)胞Sox9、Ngn3、Hes1基因表達(dá)的情況。方法:選擇17-19周齡新診斷1型糖尿病(T1D)NOD鼠24只,隨機(jī)分為A、B、C、D四組,分別代表抗CD20單抗單藥治療組、抗CD20+IL-10聯(lián)合治療組、Ad-IL-10單藥治療組及生理鹽水對(duì)照(NS)組。分別于發(fā)病后第1,3,6,9天尾靜脈注射抗CD20單抗250ug,抗CD20單抗250ug+Ad-IL-10 100ul,Ad-IL-10 100ul,NS 100ul,每3天1次,首劑加倍,共注射4次。期間每天同一時(shí)間監(jiān)測(cè)小鼠血糖水平,若血糖≥25mmol/L,給予20U/kg皮下注射來得時(shí);每周同一時(shí)間測(cè)量2次體重,并記錄。自最后1次給藥后,連續(xù)觀察9周,處死小鼠。摘眼球后取血,靜置離心取血清,行ELISA檢測(cè)。解剖分離肝臟,部分行Western Blot和RT-PCR檢測(cè);部分用4%甲醛固定,制作石蠟切片,行免疫組化檢測(cè)。結(jié)果:動(dòng)態(tài)血糖監(jiān)測(cè)示聯(lián)合治療可明顯降低NOD鼠血糖水平。動(dòng)態(tài)體質(zhì)量監(jiān)測(cè)示各組干預(yù)對(duì)NOD鼠體質(zhì)量影響不大,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。RT-PCR結(jié)果示聯(lián)合組小鼠肝臟Sox9、Ngn3基因表達(dá)明顯高于單藥治療組和生理鹽水組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),Hes1基因表達(dá)量在各組間差別不大,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);Western Blot示聯(lián)合治療組IL-10蛋白表達(dá)水平明顯高于其它組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),Hes1蛋白表達(dá)量在各組間差別不大,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);免疫熒光結(jié)果示胰島素表達(dá)細(xì)胞在聯(lián)合治療組明顯增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05);ELISA結(jié)果示聯(lián)合治療組較單藥治療組血清IL-17、IL-1β明顯減低,聯(lián)合治療組血清TGF-β、Insulin水平較其他三組明顯較高,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:腺病毒介導(dǎo)的IL-10基因在肝臟組織內(nèi)高表達(dá)�？笴D20單克隆抗體與腺病毒介導(dǎo)的IL-10聯(lián)合應(yīng)用可促進(jìn)NOD鼠胰島素的分泌,更好的維持血糖穩(wěn)定。聯(lián)合干預(yù)可以減低IL-17、IL-1β等炎性因子水平,促進(jìn)TGF-β合成,維持免疫穩(wěn)態(tài)。同時(shí),聯(lián)合干預(yù)可以通過激活Notch信號(hào)通路,促進(jìn)Sox9、Ngn3基因表達(dá)等方式促進(jìn)肝臟合成IPC(胰島內(nèi)分泌細(xì)胞),分泌肝源性胰島素,對(duì)殘存胰島β細(xì)胞功能具有一定的保護(hù)作用。
[Abstract]:Objective: to observe the expression of CD20 monoclonal antibody combined with adenovirus mediated IL-10 gene in liver cells of nonobese diabetic rats. Methods: Twenty-four newly diagnosed type 1 diabetic T1D nod mice were selected from 17 to 19 weeks old. They were randomly divided into four groups: group A, B, C, D, representing the treatment group of anti CD20 McAb, respectively. Anti CD20 IL-10 combined with Ad-IL-10 monotherapy group and normal saline control group were injected intravenously with anti-#en2# McAb 250ugand anti-#en3# McAb 250ug Ad-IL-10 100uln NS 100U every 3 days at the end of 9 days after the onset of the disease, and the first dose was doubled. The blood glucose level of mice was monitored at the same time every day. If the blood glucose was more than 25 mmol / L, 20 U / kg subcutaneous injection was given to the mice, and the weight was measured twice a week at the same time, and recorded. The mice were sacrificed, blood was taken from eyeball, serum was collected by static centrifugation, ELISA was detected, liver was dissected, Western Blot and RT-PCR were detected partly, paraffin sections were made by fixed with 4% formaldehyde. Results: dynamic blood glucose monitoring showed that combined treatment could significantly reduce the blood glucose level of NOD mice. Dynamic body mass monitoring showed that the intervention of each group had little effect on the body mass of NOD mice. The results of RT-PCR showed that the expression of Ngn3 gene in liver of combined group was significantly higher than that of single drug treatment group and normal saline group. The expression of IL-10 protein in the combined treatment group was significantly higher than that in the other groups, and there was no significant difference in the expression of Hes1 protein between the two groups. The results of immunofluorescence showed that the expression of insulin cells in the combined treatment group was significantly increased, and the difference was statistically significant. The results of Elisa showed that the serum levels of IL-17 and IL-1 尾 in the combined treatment group were significantly lower than those in the single drug treatment group. The serum TGF- 尾 insulin level in the combined treatment group was significantly higher than that in the other three groups. Conclusion: adenovirus-mediated IL-10 gene is highly expressed in liver tissue. The combination of anti-adenovirus monoclonal antibody and adenovirus-mediated IL-10 can promote insulin secretion in NOD mice. The combined intervention can reduce the level of inflammatory factors such as IL-17 and IL-1 尾, promote the synthesis of TGF- 尾 and maintain the immune homeostasis. At the same time, the combined intervention can activate the Notch signaling pathway. Promoting the expression of Sox9 Ngn3 gene and promoting the synthesis of IPCs (islet endocrine cells, secreting hepatogenic insulin, etc.) has a certain protective effect on the function of residual islet 尾 cells.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R587.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉小會(huì);;脂肪間充質(zhì)干細(xì)胞移植治療1型糖尿病模型大鼠的作用[J];中國(guó)組織工程研究;2016年41期

2 武繼承;郭林峰;唐道琪;田md;姚文兵;;GLP-1受體激動(dòng)劑的臨床藥效與安全性研究進(jìn)展[J];藥物生物技術(shù);2016年04期

3 馬建華;;1型糖尿病強(qiáng)化治療與胰島素泵應(yīng)用技巧[J];中國(guó)實(shí)用內(nèi)科雜志;2016年07期

4 唐珊珊;馬東慎;高華山;趙月葵;曹順秀;金亮;;干細(xì)胞在1型糖尿病治療中的應(yīng)用研究進(jìn)展[J];藥學(xué)進(jìn)展;2016年03期

5 閭倩;石勇銓;;維生素D與糖尿病[J];藥品評(píng)價(jià);2016年05期

6 李雅懿;;試論健康教育對(duì)糖尿病患者生活質(zhì)量的影響[J];世界最新醫(yī)學(xué)信息文摘;2016年19期

7 楊懿;高新;宋海峰;;GLP-1受體激動(dòng)劑的臨床研究進(jìn)展[J];中國(guó)新藥雜志;2015年19期

8 欽丹萍;周毅駿;張紹珠;曹俊敏;徐立宇;方國(guó)棟;王佳;;雷公藤多苷抗巨噬細(xì)胞炎癥及對(duì)TLR4/NF-κB調(diào)控炎癥作用的研究[J];中國(guó)中藥雜志;2015年16期

9 劉頤軒;臧莎莎;宋光耀;;二甲雙胍分子機(jī)制的研究進(jìn)展[J];中國(guó)糖尿病雜志;2015年04期

10 李偉;肖新華;;1型糖尿病患者的血糖監(jiān)測(cè)[J];糖尿病天地(臨床);2015年01期

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