转化生长因子α对LoVo细胞增殖及其它生物学功能影响的研究

发布时间：2018-04-05 16:07

本文选题：转化生长因子α　切入点：LoVo细胞　出处：《东北农业大学》2008年硕士论文

【摘要】： 目的:转化生长因子α(TGF-α)是由50个氨基酸残基构成的一类小分子多肽,其广泛存在于人体多种器官和体液中,研究表明TGF-α对人体胃肠道功能起着重要的调节作用,能够刺激人体胃肠道细胞的生长和分化,对胃肠道粘膜损伤具有修复和保护的功能,并可促进新生儿胃肠道的生长发育。本课题旨在研究人初乳、常乳及婴儿配方乳源性及重组人转化生长因子α(TGF-α)对人结肠上皮细胞LoVo促增殖作用,以及不同浓度外源性TGF-α对LoVo细胞RNA和蛋白质含量及细胞迁移能力的影响。方法:以人结肠细胞LoVo为研究模型,采用四甲基偶氮唑盐比色法(MTT法)检测人初乳、常乳、婴儿配方乳粉和重组人TGF-α(浓度分别为0.1、1、10和100ng/mL)作用人结肠上皮细胞模型24h后细胞增殖率的变化,以及各成分添加TGF-α抗体作用24h后人结肠上皮细胞模型增殖率的变化。分别添加重组人TGF-α0.1、1(接近人乳TGF-α含量最低值)、10(接近人乳TGF-α含量最高值)、100ng/mL后,观察LoVo细胞增殖能力、细胞总RNA、总蛋白质含量以及细胞迁移能力的变化。结果:各种成分乳以及不同浓度重组人转化生长因子α均能够显著提高人结肠上皮细胞增殖率。并且随着TGF-α浓度的升高细胞增殖程度也相应得到提高。而TGF-α抗体的中和作用使得重组人TGF-α对肠上皮细胞促增殖作用在四个浓度组中分别降低了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),同时也使得人初乳、常乳以及婴儿配方乳粉对肠上皮细胞促增殖作用分别降低了56±13%(P＜0.01)、35±3%(P＜0.05)和9±3%(P＞0.05)。添加TGF-α24小时后,细胞较对照组有明显的增殖现象,且细胞增殖率随细胞浓度增加而呈上升趋势,并在10ng/mL时达到最大(P＜0.01)。添加TGF-α48小时后,细胞增殖率依然随TGF-α浓度增大而依次升高,且在100ng/mL时达到最大(P＜0.01)。而48h与24h相比细胞增殖率有所提高,说明增殖率与TGF-α成一定的时间依赖关系。细胞总RNA含量随TGF-α浓度增加而上升为控制组的1.67倍(P＜0.05)至4.20倍(P＜0.01)。而100ng/mL组细胞总蛋白质含量明显高于控制组和其它各剂量组。为对照组的1.28倍(P＜0.05)。另外,添加重组人TGF-α与对照组相比可明显提高LoVo细胞迁移能力。结论:TGF-α抗体对含有人初乳、常乳和基于牛乳成分的婴儿配方乳粉以及重组人TGF-α培养基的中和作用使得它们对人结肠上皮细胞的促增殖作用显著降低,证明不同源性的TGF-α都能够对人体外肠道上皮细胞起到明显的促增殖作用。外源性TGF-α同样能够促进人结肠上皮细胞RNA含量的增加和蛋白质的合成。并且提高LoVo细胞的迁移能力。
[Abstract]:Objective: transforming growth factor 伪 (TGF- 伪) 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- 伪 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 伪 (TGF- 伪) on the proliferation of human colon epithelial cells (LoVo).The effects of different concentrations of exogenous TGF- 伪 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- 伪 (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- 伪 antibody.After adding recombinant human TGF- 伪 0.1 ~ (-1) (close to the lowest value of human milk TGF- 伪) (close to 100 ng / mL) of human milk TGF- 伪 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 伪 could significantly increase the proliferation rate of human colon epithelial cells.With the increase of TGF- 伪 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 卤13 (P < 0.01) and 35 卤3 (P < 0.05) and 9 卤3 (P > 0.05), respectively.When TGF- 伪 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- 伪 addition, the cell proliferation rate increased with the increase of TGF- 伪 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- 伪.With the increase of TGF- 伪 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- 伪 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- 伪 medium can significantly reduce the proliferation of human colonic epithelial cells.The results showed that TGF- 伪 with different homology could significantly promote the proliferation of human intestinal epithelial cells.Exogenous TGF- 伪 also promoted the increase of RNA content and protein synthesis in human colon epithelial cells.And improve the migration ability of LoVo cells.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R346

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