中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
27期
5336-5340
,共5页
曾祥一%王伟%孙亮%张力%曾令达
曾祥一%王偉%孫亮%張力%曾令達
증상일%왕위%손량%장력%증령체
肌源性干细胞%神经元%睫状神经营养因子%丹参注射液%核因子kB
肌源性榦細胞%神經元%睫狀神經營養因子%丹參註射液%覈因子kB
기원성간세포%신경원%첩상신경영양인자%단삼주사액%핵인자kB
背景:在干细胞异体移植时神经细胞的低存活率成为异体细胞移植失败的主要诱因.核因子kB是细胞信号传导的主要转录因子之一,参与细胞的增殖和分化过程.目的:观察睫状神经营养因子和丹参注射液联合诱导肌源性干细胞向神经元样细胞的分化情况,以及分化过程中核因子kB的表达.设计、时间及地点:细胞学基因水平实验,于2008-03/05在辽宁医学院附属第二医院口腔科学实验室完成.材料:新生7 d龄SD乳鼠10只,由辽宁医学院实验动物中心提供.睫状神经营养因子为Sigma公司产品,丹参注射液购自正大青春宝药业有限公司.方法:体外分离培养鼠肌源性干细胞,差速贴壁法和酶消化法纯化后接种于6孔板内.诱导组用含睫状神经营养因子的DMEM预诱导24 h,更换培养液,洗涤3次,再加入含丹参注射液的无血清DMEM诱导5 h;对照组使用无血清DMEM培养基处理.主要观察指标:RT-PCR及Western blotting法检测神经丝蛋白、核因子kB抑制蛋白的表达.结果:诱导前肌源性干细胞未见神经丝蛋白表达,诱导后的神经元样细胞神经丝蛋白呈阳性表达.凝胶电泳及Westem blot检测结果显示,与诱导前比较,诱导后对照组核因子kB抑制蛋白的表达无明显变化,诱导组核因子kB抑制蛋白的表达明显下降.结论:睫状神经营养因子和丹参注射液可联合诱导肌源性干细胞分化为神经元样细胞,分化过程中核因子kB的活化被抑制.
揹景:在榦細胞異體移植時神經細胞的低存活率成為異體細胞移植失敗的主要誘因.覈因子kB是細胞信號傳導的主要轉錄因子之一,參與細胞的增殖和分化過程.目的:觀察睫狀神經營養因子和丹參註射液聯閤誘導肌源性榦細胞嚮神經元樣細胞的分化情況,以及分化過程中覈因子kB的錶達.設計、時間及地點:細胞學基因水平實驗,于2008-03/05在遼寧醫學院附屬第二醫院口腔科學實驗室完成.材料:新生7 d齡SD乳鼠10隻,由遼寧醫學院實驗動物中心提供.睫狀神經營養因子為Sigma公司產品,丹參註射液購自正大青春寶藥業有限公司.方法:體外分離培養鼠肌源性榦細胞,差速貼壁法和酶消化法純化後接種于6孔闆內.誘導組用含睫狀神經營養因子的DMEM預誘導24 h,更換培養液,洗滌3次,再加入含丹參註射液的無血清DMEM誘導5 h;對照組使用無血清DMEM培養基處理.主要觀察指標:RT-PCR及Western blotting法檢測神經絲蛋白、覈因子kB抑製蛋白的錶達.結果:誘導前肌源性榦細胞未見神經絲蛋白錶達,誘導後的神經元樣細胞神經絲蛋白呈暘性錶達.凝膠電泳及Westem blot檢測結果顯示,與誘導前比較,誘導後對照組覈因子kB抑製蛋白的錶達無明顯變化,誘導組覈因子kB抑製蛋白的錶達明顯下降.結論:睫狀神經營養因子和丹參註射液可聯閤誘導肌源性榦細胞分化為神經元樣細胞,分化過程中覈因子kB的活化被抑製.
배경:재간세포이체이식시신경세포적저존활솔성위이체세포이식실패적주요유인.핵인자kB시세포신호전도적주요전록인자지일,삼여세포적증식화분화과정.목적:관찰첩상신경영양인자화단삼주사액연합유도기원성간세포향신경원양세포적분화정황,이급분화과정중핵인자kB적표체.설계、시간급지점:세포학기인수평실험,우2008-03/05재료녕의학원부속제이의원구강과학실험실완성.재료:신생7 d령SD유서10지,유료녕의학원실험동물중심제공.첩상신경영양인자위Sigma공사산품,단삼주사액구자정대청춘보약업유한공사.방법:체외분리배양서기원성간세포,차속첩벽법화매소화법순화후접충우6공판내.유도조용함첩상신경영양인자적DMEM예유도24 h,경환배양액,세조3차,재가입함단삼주사액적무혈청DMEM유도5 h;대조조사용무혈청DMEM배양기처리.주요관찰지표:RT-PCR급Western blotting법검측신경사단백、핵인자kB억제단백적표체.결과:유도전기원성간세포미견신경사단백표체,유도후적신경원양세포신경사단백정양성표체.응효전영급Westem blot검측결과현시,여유도전비교,유도후대조조핵인자kB억제단백적표체무명현변화,유도조핵인자kB억제단백적표체명현하강.결론:첩상신경영양인자화단삼주사액가연합유도기원성간세포분화위신경원양세포,분화과정중핵인자kB적활화피억제.
BACKGROUND: The low survival rate of neuron cells is one of the main mechanisms of stem cell allograft, which might lead to the failure of allograft. Nuclear factor-KB (NF-kB) is one of main transcription factors for cell signaling transduction and participates in call proliferation and differentiation.OBJECTIVE: To study the differentiation of muscle-derived stem cells (MDSCs) into neuron-like cells induced by ciliary neurotrophic factor (CNTF) and Compound Salvia Miltiorrhiza Injection in vitro, and the expression of NF- kB.DESIGN, TIME AND SETTING: The experiment at cell molecular level was performed at the Oral Science Laboratory of the Second Affiliated Hospital of Liaoning Medical College from March to May 2008.MATERIALS: A total of 10 Sprague Dawley neonatal rats aged 7 days were supplied by Experimental Animal Center, Liaoning Medical University. CNTF (Sigma, USA) and Compound Salvia Miltiorrhiza Injection (Chiatai Qingchunbao Pharmacantical Co.,Ltd., China) were used in this study.METHODS: Rat MDSCs were harvested in vitro, pudfied by differential adherence and enzyme digestion, and incubated in 6-well plate. Samples in the induction group were incubated in DMEM containing CNTF for 24 hours. The medium was changed.Subsequently, samples were dnsed three times, and then incubated in serum-free DMEM supplemented with Compound Salvia Miltiorrhiza Injection for 5 hours. Samples in the control group were treated with serum-free DMEM.MAIN OUTCOME MEASURES: Neurofilament protein and NF-KB inhibitor protein expression were detected using reverse transcription-polymerase chain reaction (RT-PCR) and Westem blotting.RESULTS: No neurofilament protein expression was found in MDSCs before induction, and neurofilament protein-positive MDSCs were detected following induction. Results of gel electrophoresis and Westam blot showed that no significant differences in NF-kB inhibitor protein expression were determined in the control group, and NF-kB inhibitor protein expression was significantly decreased in the induction group after induction.CONCLUSION: CNTF and Compound Salvia Miltiorrhiza Injection could inhibit the activation of NF-kB and induce the differentiation of MDSCs into neuron-like cells.Zeng XY, Wang W, Sun L, Zhang L, Zeng LD.Differentiation of muscle-derived stem cells into neuron-like cells induced by ciliary neurotrophic factor and Compound Salvia Miltiorrhiza Injection in vitro.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu.2009;13(27): 5336-5340. [http://www.crter.cn http://en.zglckf.com]
