Notch信號通路在臍帶間充質干細胞治療擴張型心肌病鼠中的作用
發(fā)布時間：2018-09-13 15:43
【摘要】：目的探究Notch通路在人臍帶間充質干細胞治療擴張型心肌病鼠中的作用。方法1、人臍帶間充質干細胞對γ-分泌酶抑制劑誘導的擴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(簡稱地鼠)60只,隨機分為正常組(15只)、實驗組(分為DAPT組、DAPT對照組、干細胞組,各15只)。其中實驗組地鼠每天進行腹腔注射γ-分泌酶抑制劑(DAPT)構建擴張型心肌病(DCM)模型,同時每周分別向DAPT對照組(即DMEM組)和干細胞組肌肉注射干細胞重懸培養(yǎng)液DMEM和人臍帶間充質干細胞(h HUCMSCs),應用超聲心動圖和血漿BNP水平評估地鼠心功能情況,采用HE染色和Masson染色觀察心肌組織變化情況。2、檢測人臍帶間充質干細胞治療擴張型心肌病大鼠后心肌組織Notch信號通路的活性110只SPF級健康、雄性SD大鼠隨機分為正常組(20只)、擴張型心肌病組(簡稱DCM組,90只)。DCM組大鼠持續(xù)8周、每周1次經(jīng)腹腔注射阿霉素構建擴張型心肌病模型,將建模成功后的60只DCM組大鼠隨機分為DCM對照組(簡稱DMEM組)、h HUCMSCs低劑量組(簡稱低劑量組)、h HUCMSCs高劑量組(簡稱高劑量組),分別經(jīng)肌肉注射DMEM、低劑量干細胞和高劑量干細胞治療。h HUCMSC治療2周及4周后,應用超聲心動圖和血漿BNP水平評估地鼠心功能情況,采用Western Blot檢測心肌組織NICD(或者N1ICD,Notch信號通路游離的胞內段)和Notch信號通路下游因子Hes1的蛋白含量。結果1、人臍帶間充質干細胞對γ-分泌酶抑制劑誘導的擴張型心肌病敘利亞金黃地鼠的治療作用超聲心動圖結果,注射藥物前各組差異無統(tǒng)計學意義(P0.05);注射藥物后DAPT組、DMEM組、干細胞組與正常組相比差異有統(tǒng)計學意義(P0.01),DAPT組、DMEM組與干細胞組相比差異有統(tǒng)計學意義(P0.05),而DAPT組和DMEM組無顯著差異(P0.05)。血漿BNP水平,注射藥物后DAPT組、DMEM組與正常組、干細胞組相比差異有統(tǒng)計學意義(P0.01),而干細胞組較正常組無顯著差異(P0.05)。HE染色顯示,與DAPT組和DMEM組相比,正常組和干細胞組心肌細胞水腫減輕,炎性細胞浸潤減少,心肌纖維排列較整齊;Masson染色顯示干細胞組心肌間藍染的膠原纖維沉積減少。2、人臍帶間充質干細胞治療擴張型心肌病鼠心肌組織中Notch信號通路活性正常組大鼠治療前、治療2周及4周后超聲心動圖結果無顯著差異(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圖超聲結果無顯著差異(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檢測結果顯示治療4周后各組之間心肌組織中NICD、Hes1蛋白含量差異無統(tǒng)計學意義(P0.05)。結論1、h UCMSCs可有效改善Notch信號通路被抑制后引起的擴張型心肌病地鼠的心功能;2、阿霉素誘導的擴張型心肌病大鼠Notch信號通路活性較正常組無顯著改變,肌肉注射h UCMSCs可明顯改善心功能,但對心肌組織Notch信號通路的活性無顯著影響。以上結果提示h UCMSCs對擴張型心肌病的修復作用不依賴于對Notch信號通路的調節(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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R725.4
本文編號：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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R725.4
【參考文獻】
中國期刊全文數(shù)據(jù)庫 前7條
1 許耘紅;何穗昒;李麗青;;心臟發(fā)育相關基因NKX2.5和GATA4突變與先天性心臟病的相關性[J];中國臨床藥理學雜志;2016年22期
2 王本臻;毛成剛;李榮;寧險峰;李自普;;人臍帶間充質干細胞肌肉注射對多柔比星誘導擴張型心肌病大鼠心功能及心肌超微結構的影響[J];中華實用兒科臨床雜志;2016年01期
3 陳嘉楠;呂玉蘭;何炳興;;血漿BNP水平應用在慢性心力衰竭的診斷、病情及預后評估中的效果分析[J];吉林醫(yī)學;2015年13期
4 張文祥;王思平;錫洪敏;李自普;;Wistar大鼠肌肉注射異種臍帶間充質干細胞的安全性[J];中國組織工程研究;2014年41期
5 王琪;張文祥;王思平;李自普;;肌肉注射人臍帶間充質干細胞健康大鼠心肌組織ki-67、phh3及cTnT的表達[J];中國組織工程研究;2014年41期
6 馬錫慧;馮凱;石炳毅;;人臍帶間充質干細胞生物學特性及其研究進展[J];中國組織工程研究與臨床康復;2011年32期
7 曹青;王飛;林繼先;陳書艷;;骨髓間充質干細胞對CD117~+心臟干細胞分化過程中轉化生長因子βⅢ型受體表達的影響[J];中國組織工程研究與臨床康復;2009年36期
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