Notch信號(hào)通路在臍帶間充質(zhì)干細(xì)胞治療擴(kuò)張型心肌病鼠中的作用
發(fā)布時(shí)間：2018-09-13 15:43
【摘要】：目的探究Notch通路在人臍帶間充質(zhì)干細(xì)胞治療擴(kuò)張型心肌病鼠中的作用。方法1、人臍帶間充質(zhì)干細(xì)胞對(duì)γ-分泌酶抑制劑誘導(dǎo)的擴(kuò)張型心肌病敘利亞金黃地鼠疾病模型的治療作用雄性健康敘利亞金黃地鼠(簡(jiǎn)稱地鼠)60只,隨機(jī)分為正常組(15只)、實(shí)驗(yàn)組(分為DAPT組、DAPT對(duì)照組、干細(xì)胞組,各15只)。其中實(shí)驗(yàn)組地鼠每天進(jìn)行腹腔注射γ-分泌酶抑制劑(DAPT)構(gòu)建擴(kuò)張型心肌病(DCM)模型,同時(shí)每周分別向DAPT對(duì)照組(即DMEM組)和干細(xì)胞組肌肉注射干細(xì)胞重懸培養(yǎng)液DMEM和人臍帶間充質(zhì)干細(xì)胞(h HUCMSCs),應(yīng)用超聲心動(dòng)圖和血漿BNP水平評(píng)估地鼠心功能情況,采用HE染色和Masson染色觀察心肌組織變化情況。2、檢測(cè)人臍帶間充質(zhì)干細(xì)胞治療擴(kuò)張型心肌病大鼠后心肌組織Notch信號(hào)通路的活性110只SPF級(jí)健康、雄性SD大鼠隨機(jī)分為正常組(20只)、擴(kuò)張型心肌病組(簡(jiǎn)稱DCM組,90只)。DCM組大鼠持續(xù)8周、每周1次經(jīng)腹腔注射阿霉素構(gòu)建擴(kuò)張型心肌病模型,將建模成功后的60只DCM組大鼠隨機(jī)分為DCM對(duì)照組(簡(jiǎn)稱DMEM組)、h HUCMSCs低劑量組(簡(jiǎn)稱低劑量組)、h HUCMSCs高劑量組(簡(jiǎn)稱高劑量組),分別經(jīng)肌肉注射DMEM、低劑量干細(xì)胞和高劑量干細(xì)胞治療。h HUCMSC治療2周及4周后,應(yīng)用超聲心動(dòng)圖和血漿BNP水平評(píng)估地鼠心功能情況,采用Western Blot檢測(cè)心肌組織NICD(或者N1ICD,Notch信號(hào)通路游離的胞內(nèi)段)和Notch信號(hào)通路下游因子Hes1的蛋白含量。結(jié)果1、人臍帶間充質(zhì)干細(xì)胞對(duì)γ-分泌酶抑制劑誘導(dǎo)的擴(kuò)張型心肌病敘利亞金黃地鼠的治療作用超聲心動(dòng)圖結(jié)果,注射藥物前各組差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);注射藥物后DAPT組、DMEM組、干細(xì)胞組與正常組相比差異有統(tǒng)計(jì)學(xué)意義(P0.01),DAPT組、DMEM組與干細(xì)胞組相比差異有統(tǒng)計(jì)學(xué)意義(P0.05),而DAPT組和DMEM組無(wú)顯著差異(P0.05)。血漿BNP水平,注射藥物后DAPT組、DMEM組與正常組、干細(xì)胞組相比差異有統(tǒng)計(jì)學(xué)意義(P0.01),而干細(xì)胞組較正常組無(wú)顯著差異(P0.05)。HE染色顯示,與DAPT組和DMEM組相比,正常組和干細(xì)胞組心肌細(xì)胞水腫減輕,炎性細(xì)胞浸潤(rùn)減少,心肌纖維排列較整齊;Masson染色顯示干細(xì)胞組心肌間藍(lán)染的膠原纖維沉積減少。2、人臍帶間充質(zhì)干細(xì)胞治療擴(kuò)張型心肌病鼠心肌組織中Notch信號(hào)通路活性正常組大鼠治療前、治療2周及4周后超聲心動(dòng)圖結(jié)果無(wú)顯著差異(P0.05);h HUCMSCs移植2周后DMEM組、低劑量組及高劑量組治療前后超聲心動(dòng)圖超聲結(jié)果無(wú)顯著差異(P0.05);h HUCMSCs移植4周后DMEM組、低劑量組及高劑量組治療前后LVEDD、LVESD無(wú)顯著差異,而LVEF、LVFS差異顯著,且DMEM組LVEF、LVFS低于低劑量組和高劑量組(P0.05)。Western Blot檢測(cè)結(jié)果顯示治療4周后各組之間心肌組織中NICD、Hes1蛋白含量差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論1、h UCMSCs可有效改善Notch信號(hào)通路被抑制后引起的擴(kuò)張型心肌病地鼠的心功能;2、阿霉素誘導(dǎo)的擴(kuò)張型心肌病大鼠Notch信號(hào)通路活性較正常組無(wú)顯著改變,肌肉注射h UCMSCs可明顯改善心功能,但對(duì)心肌組織Notch信號(hào)通路的活性無(wú)顯著影響。以上結(jié)果提示h UCMSCs對(duì)擴(kuò)張型心肌病的修復(fù)作用不依賴于對(duì)Notch信號(hào)通路的調(diào)節(jié)。
[Abstract]:Objective To explore the role of Notch pathway in human umbilical cord mesenchymal stem cells (HUCMSCs) in the treatment of dilated cardiomyopathy (DCM) in mice. Methods 1. Human umbilical cord mesenchymal stem cells (HUCMSCs) were used to treat DCM induced by gamma-secretase inhibitors in Syrian golden hamsters. Sixty healthy male Syrian hamsters (hamsters) were randomly divided into normal control group and control group. The hamsters in the experimental group were injected intraperitoneally with gamma-secretase inhibitor (DAPT) every day to establish dilated cardiomyopathy (DCM) model. At the same time, the hamsters in the experimental group were injected intramuscularly with DMEM and human umbilical cord mesenchymal filling (HUCM) in the DAPT control group (DMEM) and the stem cell group respectively weekly. Cardiac function of hamsters was assessed by echocardiography and plasma BNP levels. The changes of myocardial tissue were observed by HE staining and Mason staining. 2. The Notch signaling pathway activity of myocardial tissue in dilated cardiomyopathy rats treated with human umbilical cord mesenchymal stem cells (HUCMSCs) was detected in 110 SPF healthy male SD rats. DCM rats were injected intraperitoneally with adriamycin once a week for 8 weeks to establish dilated cardiomyopathy model. 60 DCM rats were randomly divided into DCM control group (DMEM group), low dose group of hUCMSCs (low dose group) and high dose group of hUCMSCs (abbreviated as high dose group). After 2 and 4 weeks of treatment with HUCMSC, echocardiography and plasma BNP levels were used to assess the cardiac function of hamsters. Western Blot was used to detect NICD (or intracellular segment of N1ICD, Notch signaling pathway) and downstream Notch signaling pathway in myocardium. Results 1. The therapeutic effect of human umbilical cord mesenchymal stem cells on dilated cardiomyopathy induced by gamma-secretase inhibitor in Syrian golden hamsters showed no significant difference between the groups before injection (P 0.05); there was statistical difference between DAPT group, DMEM group and normal group after injection of drugs. Significance (P 0.01), DAPT group, DMEM group and stem cell group were significantly different (P 0.05), but there was no significant difference between DAPT group and DMEM group (P 0.05). Compared with DAPT group and DMEM group, myocardial edema, inflammatory cell infiltration and myocardial fiber arrangement were reduced in normal group and stem cell group. Masson staining showed that the deposition of blue-stained collagen fibers was decreased in stem cell group. 2. Notch signaling pathway was activated in human umbilical cord mesenchymal stem cells treated dilated cardiomyopathy rats. There was no significant difference in echocardiographic results between two and four weeks before and after treatment in normal rats (P 0.05); there was no significant difference in echocardiographic results between two weeks after transplantation in DMEM group, low dose group and high dose group (P 0.05); there was no significant difference in LVEDD and LVESD between four weeks after transplantation in DMEM group, low dose group and high dose group. There were significant differences in LVEF and LVFS, and LVEF and LVFS in DMEM group were lower than those in low-dose group and high-dose group (P 0.05). Western Blot test results showed that there was no significant difference in the contents of NICD and Hes1 protein between the groups after 4 weeks of treatment (P 0.05). 2. Notch signaling pathway activity of adriamycin-induced dilated cardiomyopathy rats was not significantly changed compared with the normal group. Intramuscular injection of H UCMSCs could significantly improve cardiac function, but had no significant effect on Notch signaling pathway activity of myocardial tissue. Regulation of Notch signaling pathway.
【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)】：R725.4
本文編號(hào)：2241607
[Abstract]:Objective To explore the role of Notch pathway in human umbilical cord mesenchymal stem cells (HUCMSCs) in the treatment of dilated cardiomyopathy (DCM) in mice. Methods 1. Human umbilical cord mesenchymal stem cells (HUCMSCs) were used to treat DCM induced by gamma-secretase inhibitors in Syrian golden hamsters. Sixty healthy male Syrian hamsters (hamsters) were randomly divided into normal control group and control group. The hamsters in the experimental group were injected intraperitoneally with gamma-secretase inhibitor (DAPT) every day to establish dilated cardiomyopathy (DCM) model. At the same time, the hamsters in the experimental group were injected intramuscularly with DMEM and human umbilical cord mesenchymal filling (HUCM) in the DAPT control group (DMEM) and the stem cell group respectively weekly. Cardiac function of hamsters was assessed by echocardiography and plasma BNP levels. The changes of myocardial tissue were observed by HE staining and Mason staining. 2. The Notch signaling pathway activity of myocardial tissue in dilated cardiomyopathy rats treated with human umbilical cord mesenchymal stem cells (HUCMSCs) was detected in 110 SPF healthy male SD rats. DCM rats were injected intraperitoneally with adriamycin once a week for 8 weeks to establish dilated cardiomyopathy model. 60 DCM rats were randomly divided into DCM control group (DMEM group), low dose group of hUCMSCs (low dose group) and high dose group of hUCMSCs (abbreviated as high dose group). After 2 and 4 weeks of treatment with HUCMSC, echocardiography and plasma BNP levels were used to assess the cardiac function of hamsters. Western Blot was used to detect NICD (or intracellular segment of N1ICD, Notch signaling pathway) and downstream Notch signaling pathway in myocardium. Results 1. The therapeutic effect of human umbilical cord mesenchymal stem cells on dilated cardiomyopathy induced by gamma-secretase inhibitor in Syrian golden hamsters showed no significant difference between the groups before injection (P 0.05); there was statistical difference between DAPT group, DMEM group and normal group after injection of drugs. Significance (P 0.01), DAPT group, DMEM group and stem cell group were significantly different (P 0.05), but there was no significant difference between DAPT group and DMEM group (P 0.05). Compared with DAPT group and DMEM group, myocardial edema, inflammatory cell infiltration and myocardial fiber arrangement were reduced in normal group and stem cell group. Masson staining showed that the deposition of blue-stained collagen fibers was decreased in stem cell group. 2. Notch signaling pathway was activated in human umbilical cord mesenchymal stem cells treated dilated cardiomyopathy rats. There was no significant difference in echocardiographic results between two and four weeks before and after treatment in normal rats (P 0.05); there was no significant difference in echocardiographic results between two weeks after transplantation in DMEM group, low dose group and high dose group (P 0.05); there was no significant difference in LVEDD and LVESD between four weeks after transplantation in DMEM group, low dose group and high dose group. There were significant differences in LVEF and LVFS, and LVEF and LVFS in DMEM group were lower than those in low-dose group and high-dose group (P 0.05). Western Blot test results showed that there was no significant difference in the contents of NICD and Hes1 protein between the groups after 4 weeks of treatment (P 0.05). 2. Notch signaling pathway activity of adriamycin-induced dilated cardiomyopathy rats was not significantly changed compared with the normal group. Intramuscular injection of H UCMSCs could significantly improve cardiac function, but had no significant effect on Notch signaling pathway activity of myocardial tissue. Regulation of Notch signaling pathway.
【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)】：R725.4
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