| Abstract: | 利用培養中的纖維母細胞，添加母乳的實驗中發現纖維網狀蛋白Fibronectin生合成有上升的現象，而母乳經人體的消化系統吸收代謝，首先便是接觸內皮細胞，在前年本實驗室的研究以顯示，添加母乳於培養的內皮細胞後，的確也可促進Fibronectin的生合成增加，而本實驗想進一步探討母乳中有何種物質可誘發此一現象的進行，而其可能的路徑又為何
實驗利用培養的內皮細胞，分別添加母乳(1%、5%、10%)、anti-TGF-β抗體、母乳中IgA初純化物(1、5、10μg/ml)及EGF(1、5、10ng/ml)於培養液中，培養0.5、1.0、2.0、24、48小時後，觀察細胞數目的變化，Fibronectin的濃度改變與TGF-β表現情形。
結果顯示 : 添加母乳組，在Fibronectin濃度及TGF-β表現上均比添加配方乳汁組高，而且隨著添加的濃度與時間增加而增加 ; 在添加anti-TGF-β抗體方面組，測出的Fibronectin濃度及TGF-β都與對照組相近，但似乎只在短時間有效果，尤其是同時添加母乳時更為明顯 ; 另外在添加IgA與EGF組方面，已添加IgA 10 μm/ml、EGF 10 ng/ml 表現較同組好，在反應時間上EGF在短時間效果佳，IgA的表現是隨著時間的增加而增加 ; 本實驗雖做了24、48、小時時間點的實驗，期望觀察細胞數目是否也會因母乳或其他物質的添加而改變，但可能是養分不足及配方乳汁無法滅菌而污染的因素，導致無法比較或統計這部分的數據，其他實驗方面在細胞數目及蛋白質含量上均無統計上的差異。
綜合以上實驗結果得知，母乳確實比配方乳汁會刺激內皮細胞之細胞外間質Fibronectin的生合成增加，姆乳中的主要生長因子EGF，及主要免疫物質IgA，均可刺激Fibronectin的生合成及TGF-β表現增加，或許是反應機制不同在反應時間上也不同，因而在添加anti-TGF-β抗體後，Fibronectin的生合成及TGF-β表現都有下降的現象，顯示內皮細胞被誘發Fibronectin的表現，似乎部分可由TGF-β的調節而來，至於詳細的機制與IgA和EGF表現差異上，則有待後續的實驗來證實。
In the present study, by adding human milk to culture the fibroblast, the fibronectin synthesis was found to increase. The human milk, after metabolism by human's digestion system will reach the endothelial cells first. The previous studied conducted in our labroatory have show that the endothelial cells culture by addition of milk will promote the synthesis of fibronectin. The present study aims to explore what components of induce thus phenomena what the mecchanism is
In the study, to culture endothelial cells the human milk (1、5、10%), formula (1,5,10%), anti-TGF-βantibody, IgA(1,5,10μg/ml), and EGF(1、5、10ng/ml) were added separately to the medium. After 0.5,1.0,2.0,24,48 hours, the change of the numbers of cells, concentration of fibronectin, and expression of TGF- β were determined. The result of the group of add human milk showed the concentration of fibronectin and expression of TGF-β will more than the group of added formula milk, and this was time-dependent and dose-depedent.The survery of the concentration of fibronectin and expression of TGF-β in the groups added anti-TGF-β antibody showed the similar result to that of the controls. however it seemed to work only in short time. Especially when human milk was added at the same time , the phenomena was more obvious. In the groups. AS for the factors ,EGF worked most effciently in short time and IgA was time-dependent. The present study also absented how the numbers of cells would change on the addition of human milk or other composition for 24 and 48 hours. However , due to the malnutrition and contamination of formula, the result failed to provide data for the statistics analysis and comparison . There was no significance different between the number of cells and concertration of protein in our observed.
The study illustrated the human milk stimulate synthesis and increase of endothelial cells of extracellular matrix-fibronectin better than formula. The main growth factor,EGF,and important immune complex ,IgA,of human milk could both increase the fibronectin synthsis and expression of TGF-β.However the difference mechanism caused difference in the response. The addition of the anti-TGF-β would decrease the synthesis of fibronectin and expression of TGF-β. In mean that the increase of fibronectin would be partially regulate by TGF-β. Details of the mechanism and the difference IgA and EGF expression need further research to explore. |
