本文選題：轉(zhuǎn)化生長(zhǎng)因子α　切入點(diǎn)：LoVo細(xì)胞　出處：《東北農(nóng)業(yè)大學(xué)》2008年碩士論文

【摘要】： 目的:轉(zhuǎn)化生長(zhǎng)因子α(TGF-α)是由50個(gè)氨基酸殘基構(gòu)成的一類(lèi)小分子多肽,其廣泛存在于人體多種器官和體液中,研究表明TGF-α對(duì)人體胃腸道功能起著重要的調(diào)節(jié)作用,能夠刺激人體胃腸道細(xì)胞的生長(zhǎng)和分化,對(duì)胃腸道粘膜損傷具有修復(fù)和保護(hù)的功能,并可促進(jìn)新生兒胃腸道的生長(zhǎng)發(fā)育。 本課題旨在研究人初乳、常乳及嬰兒配方乳源性及重組人轉(zhuǎn)化生長(zhǎng)因子α(TGF-α)對(duì)人結(jié)腸上皮細(xì)胞LoVo促增殖作用,以及不同濃度外源性TGF-α對(duì)LoVo細(xì)胞RNA和蛋白質(zhì)含量及細(xì)胞遷移能力的影響。 方法:以人結(jié)腸細(xì)胞LoVo為研究模型,采用四甲基偶氮唑鹽比色法(MTT法)檢測(cè)人初乳、常乳、嬰兒配方乳粉和重組人TGF-α(濃度分別為0.1、1、10和100ng/mL)作用人結(jié)腸上皮細(xì)胞模型24h后細(xì)胞增殖率的變化,以及各成分添加TGF-α抗體作用24h后人結(jié)腸上皮細(xì)胞模型增殖率的變化。 分別添加重組人TGF-α0.1、1(接近人乳T(mén)GF-α含量最低值)、10(接近人乳T(mén)GF-α含量最高值)、100ng/mL后,觀察LoVo細(xì)胞增殖能力、細(xì)胞總RNA、總蛋白質(zhì)含量以及細(xì)胞遷移能力的變化。 結(jié)果:各種成分乳以及不同濃度重組人轉(zhuǎn)化生長(zhǎng)因子α均能夠顯著提高人結(jié)腸上皮細(xì)胞增殖率。并且隨著TGF-α濃度的升高細(xì)胞增殖程度也相應(yīng)得到提高。而TGF-α抗體的中和作用使得重組人TGF-α對(duì)腸上皮細(xì)胞促增殖作用在四個(gè)濃度組中分別降低了11±3.2%(P＜0.05)、12±0.3%(P＜0.05)、3±2.1%(P＞0.05)和4±3.3%(P＞0.05),同時(shí)也使得人初乳、常乳以及嬰兒配方乳粉對(duì)腸上皮細(xì)胞促增殖作用分別降低了56±13%(P＜0.01)、35±3%(P＜0.05)和9±3%(P＞0.05)。 添加TGF-α24小時(shí)后,細(xì)胞較對(duì)照組有明顯的增殖現(xiàn)象,且細(xì)胞增殖率隨細(xì)胞濃度增加而呈上升趨勢(shì),并在10ng/mL時(shí)達(dá)到最大(P＜0.01)。添加TGF-α48小時(shí)后,細(xì)胞增殖率依然隨TGF-α濃度增大而依次升高,且在100ng/mL時(shí)達(dá)到最大(P＜0.01)。而48h與24h相比細(xì)胞增殖率有所提高,說(shuō)明增殖率與TGF-α成一定的時(shí)間依賴(lài)關(guān)系。 細(xì)胞總RNA含量隨TGF-α濃度增加而上升為控制組的1.67倍(P＜0.05)至4.20倍(P＜0.01)。而100ng/mL組細(xì)胞總蛋白質(zhì)含量明顯高于控制組和其它各劑量組。為對(duì)照組的1.28倍(P＜0.05)。另外,添加重組人TGF-α與對(duì)照組相比可明顯提高LoVo細(xì)胞遷移能力。 結(jié)論:TGF-α抗體對(duì)含有人初乳、常乳和基于牛乳成分的嬰兒配方乳粉以及重組人TGF-α培養(yǎng)基的中和作用使得它們對(duì)人結(jié)腸上皮細(xì)胞的促增殖作用顯著降低,證明不同源性的TGF-α都能夠?qū)θ梭w外腸道上皮細(xì)胞起到明顯的促增殖作用。 外源性TGF-α同樣能夠促進(jìn)人結(jié)腸上皮細(xì)胞RNA含量的增加和蛋白質(zhì)的合成。并且提高LoVo細(xì)胞的遷移能力。
[Abstract]:Objective: transforming growth factor 偽 (TGF- 偽) is a kind of small polypeptide consisting of 50 amino acid residues, which is widely found in many organs and body fluids. Studies have shown that TGF- 偽 plays an important role in regulating gastrointestinal function.It can stimulate the growth and differentiation of human gastrointestinal cells, repair and protect gastrointestinal mucosal injury, and promote the growth and development of neonatal gastrointestinal tract.The aim of this study was to investigate the effects of human colostrum, normal milk and infant formula milk and recombinant human transforming growth factor 偽 (TGF- 偽) on the proliferation of human colon epithelial cells (LoVo).The effects of different concentrations of exogenous TGF- 偽 on the content of RNA and protein and the migration ability of LoVo cells were studied.Methods: human colostrum and normal milk were detected by tetramethylazolium colorimetric assay using human colon cell LoVo as the study model.Effects of infant formula milk powder and recombinant human TGF- 偽 (0.1 mg / mL) on the proliferation rate of human colonic epithelial cell model after 24h, and the changes of proliferation rate of human colonic epithelial cell model after 24 h exposure to TGF- 偽 antibody.After adding recombinant human TGF- 偽 0.1 ~ (-1) (close to the lowest value of human milk TGF- 偽) (close to 100 ng / mL) of human milk TGF- 偽 content, the proliferation ability, total RNAs, total protein content and cell migration ability of LoVo cells were observed.Results: all kinds of milk and different concentrations of recombinant human transforming growth factor 偽 could significantly increase the proliferation rate of human colon epithelial cells.With the increase of TGF- 偽 concentration, the cell proliferation was also increased.The effects of normal milk and infant formula milk powder on the proliferation of intestinal epithelial cells were decreased by 56 鹵13 (P < 0.01) and 35 鹵3 (P < 0.05) and 9 鹵3 (P > 0.05), respectively.When TGF- 偽 was added for 24 hours, the proliferation of the cells was more obvious than that of the control group, and the cell proliferation rate increased with the increase of the cell concentration, and reached the maximum at the time of 10ng/mL (P < 0.01).After 48 hours of TGF- 偽 addition, the cell proliferation rate increased with the increase of TGF- 偽 concentration, and reached the maximum at 100ng/mL (P < 0.01).The cell proliferation rate increased at 48 h and 24 h, indicating that the proliferation rate was in a time-dependent relationship with TGF- 偽.With the increase of TGF- 偽 concentration, the total RNA content increased to 1.67 times of control group (P < 0.05) to 4.20 times (P < 0.01).The total protein content in 100ng/mL group was significantly higher than that in control group and other dose groups.It was 1.28 times of the control group (P < 0.05).In addition, addition of recombinant human TGF- 偽 significantly increased the migration ability of LoVo cells compared with the control group.Conclusion the neutralization of the antibody against human colostrum, normal milk, infant formula milk based on bovine milk and recombinant human TGF- 偽 medium can significantly reduce the proliferation of human colonic epithelial cells.The results showed that TGF- 偽 with different homology could significantly promote the proliferation of human intestinal epithelial cells.Exogenous TGF- 偽 also promoted the increase of RNA content and protein synthesis in human colon epithelial cells.And improve the migration ability of LoVo cells.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類(lèi)號(hào)】：R346

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