本文關(guān)鍵詞： 植酸 結(jié)直腸癌 機(jī)械屏障　出處：《青島大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的:通過(guò)建立二甲肼(DMH)誘導(dǎo)的結(jié)直腸癌大鼠模型,觀察不同劑量植酸干預(yù)對(duì)結(jié)直腸癌大鼠腸粘膜機(jī)械屏障粘液層MUC2和TFF3蛋白及三種緊密連接蛋白的影響,探討植酸對(duì)結(jié)直腸癌大鼠模型腸粘膜機(jī)械屏障的作用機(jī)制。方法:60只Wistar雄性大鼠按體重隨機(jī)區(qū)組分為正常對(duì)照組(NC)、模型組(MG)、植酸低劑量組(PL)、植酸中劑量組(PM)和植酸高劑量組(PH)。各植酸作用組每天分別給予0.25g/kg、0.5g/kg和1g/kg的植酸鈉灌胃,NC和MG組灌胃等體積生理鹽水。實(shí)驗(yàn)第5周起,除NC組外,其余各組大鼠頸背部注射劑量為30mg/kg的DMH持續(xù)18周,期間NC組注射等體積生理鹽水。繼續(xù)植酸鈉灌胃至實(shí)驗(yàn)39周,處死大鼠。取NC組結(jié)腸組織及其他組結(jié)直腸腫瘤,免疫組化的方法檢測(cè)其zo-1和occludin蛋白的表達(dá)情況;Western Blot方法檢測(cè)MUC2、TFF3和claudin-1蛋白的表達(dá)情況;實(shí)時(shí)定量PCR方法檢測(cè)MUC2、TFF3和claudin-1 m RNA的表達(dá)情況;酶聯(lián)免疫吸附法(ELISA)檢測(cè)血清中D-乳酸(D-LA)和內(nèi)毒素(LPS)的表達(dá)情況。結(jié)果:實(shí)驗(yàn)結(jié)束時(shí),模型組和植酸各劑量組均發(fā)生結(jié)直腸腫瘤,其中,模型組與植酸低劑量組腫瘤發(fā)生率均為100%,植酸中、高劑量組腫瘤發(fā)生率下降,分別為88.9%和66.7%;與對(duì)照組比較,模型組zo-1和occludin蛋白的表達(dá)明顯降低(P0.05),植酸干預(yù)后,結(jié)直腸腫瘤中zo-1和occludin蛋白的表達(dá)顯著升高(P0.05);DMH誘導(dǎo)使得MUC2和TFF3蛋白表達(dá)明顯降低(P0.05),claudin-1蛋白表達(dá)明顯升高(P0.05),經(jīng)植酸作用,各植酸組MUC2蛋白表達(dá)均明顯升高(P0.05),植酸中、高劑量組TFF3的蛋白表達(dá)明顯升高(P0.05),且植酸中、高劑量組claudin-1蛋白的表達(dá)顯著降低(P0.05);MUC2和claudin-1 m RNA表達(dá)水平的變化趨勢(shì)與其對(duì)應(yīng)的蛋白變化一致,而各植酸作用組TFF3 m RNA的表達(dá)水平與模型組相比均明顯升高(P0.05)。與對(duì)照組相比,模型組大鼠血清中D-LA和LPS的表達(dá)明顯升高(P0.05),各植酸作用組顯著降低D-LA和LPS的表達(dá)(P0.05)。結(jié)論:(1).植酸可以改善DMH誘導(dǎo)的腸粘膜損傷,保護(hù)腸粘膜機(jī)械屏障,減少DMH誘導(dǎo)的大鼠結(jié)直腸腫瘤的發(fā)生率。(2)植酸通過(guò)上調(diào)腸粘膜粘液層中MUC2和TFF3蛋白及m RNA的表達(dá)水平來(lái)改善粘液層的屏障功能;植酸可以上調(diào)腸粘膜緊密連接蛋白zo-1、occludin的水平,并下調(diào)claudin-1的水平,進(jìn)而調(diào)節(jié)腸上皮細(xì)胞的連接功能;植酸下調(diào)了血清中D-LA和LPS的水平,減少機(jī)體遭受侵襲及移位的可能。
[Abstract]:Aim: to establish a rat model of colorectal cancer induced by dimethylhydrazide (DMH), and to observe the effects of different doses of phytic acid on the mucus layer MUC2, TFF3 and three kinds of tight junction proteins in the mucous layer of intestinal mucosal barrier in rats with colorectal cancer. To investigate the mechanism of phytic acid on intestinal mucosal mechanical barrier in colorectal cancer rat model. Methods 60 Wistar male rats were randomly divided into normal control group, model group, low dose group, phytic acid medium dose group and middle dose group. Each phytic acid group was given 0.25 g / kg of 0.5 g / kg and 1 g / kg of sodium phytate by intragastric administration of normal saline of the same volume as NC group and MG group respectively from the 5th week of the experiment. Except NC group, the other groups were injected with 30 mg / kg DMH for 18 weeks, during which the NC group was injected with normal saline of the same volume. The rats were sacrificed after oral administration of sodium phytate for 39 weeks. The colon tissue of NC group and other groups of colorectal tumor were taken out. The expression of zo-1 and occludin protein was detected by immunohistochemical method, the expression of MUC2tFF3 and claudin-1 protein was detected by Western Blot, the expression of MUC2tF3 and claudin-1 m RNA was detected by real-time quantitative PCR method. Results: at the end of the experiment, colorectal neoplasms were found in the model group and each dose group of phytic acid, in which, the expression of D- lactic acid D-LAA and endotoxin LPS) in serum was detected by ELISA.Results: at the end of the experiment, colorectal neoplasms were found in the model group and the phytic acid group. The incidence of tumor in the model group and the low dose group was 100. In the phytic acid group, the tumor incidence rate in the high dose group was 88.9% and 66.7, respectively. Compared with the control group, the expression of zo-1 and occludin protein in the model group was significantly lower than that in the control group, and after the intervention of phytic acid, the expression of zo-1 and occludin protein in the model group was significantly lower than that in the control group. The expression of zo-1 and occludin in colorectal neoplasms was significantly increased. The expression of MUC2 and TFF3 protein decreased significantly in colorectal neoplasms. After phytic acid treatment, the expression of MUC2 protein in each phytic acid group was significantly increased, and in phytic acid, the expression of MUC2 protein was significantly increased in phytic acid. The protein expression of TFF3 increased significantly in high dose group, and in phytic acid, the expression of claudin-1 protein in high dose group decreased significantly the changes of MUC2 and claudin-1 m RNA levels in high dose group were consistent with the corresponding protein changes. The expression of TFF3 m RNA in each phytic acid group was significantly higher than that in the model group. The expression of D-LA and LPS in serum of the model group was significantly increased, and the expression of D-LA and LPS was significantly decreased in each phytic acid group. Conclusion the expression of D-LA and LPS was significantly decreased in the model group. Phytic acid can improve the intestinal mucosal injury induced by DMH and protect the intestinal mucosal mechanical barrier. To reduce the incidence of colorectal neoplasms induced by DMH) phytic acid can improve the barrier function of mucous layer by up-regulating the expression of MUC2 and TFF3 protein and m RNA in mucous layer of intestine, and phytic acid can up-regulate the level of zo-1occludin in intestinal mucosal mucous layer. Phytic acid down-regulated the levels of D-LA and LPS in serum and reduced the possibility of invasion and translocation.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R735.3;R-332

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 Amanda M Pattison;Dante J Merlino;Erik S Blomain;Scott A Waldman;;Guanylyl cyclase C signaling axis and colon cancer prevention[J];World Journal of Gastroenterology;2016年36期

2 Zheng Zhang;Zhihua Liu;;Paneth cells: the hub for sensing and regulating intestinal flora[J];Science China(Life Sciences);2016年05期

3 葉毅君;陸虞蓀;周雪;陳卉怡;薛世勇;蒯樂(lè);;針刺對(duì)5-氟尿嘧啶所致腸黏膜屏障破壞大鼠的保護(hù)作用[J];針刺研究;2016年02期

4 Hai-Xia Shang;An-Qi Wang;Chun-Hui Bao;Huan-Gan Wu;Wei-Feng Chen;Lu-Yi Wu;Rong Ji;Ji-Meng Zhao;Yin Shi;;Moxibustion combined with acupuncture increases tight junction protein expression in Crohn's disease patients[J];World Journal of Gastroenterology;2015年16期

5 Lena Antoni;Sabine Nuding;Jan Wehkamp;Eduard F Stange;;Intestinal barrier in inflammatory bowel disease[J];World Journal of Gastroenterology;2014年05期

6 王金鳳;陳慶森;閆亞麗;;酪蛋白糖巨肽對(duì)大鼠腸黏膜屏障的保護(hù)作用[J];食品科學(xué);2013年07期

7 ;Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques[J];World Journal of Gastroenterology;2004年20期

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本文編號(hào)：1537768