缺血再灌注小腸中DNA甲基化對細胞凋亡和增殖的調控△_《中國普外基礎與臨床雜志》

作者：

 安樹才 ^1,2 , 王慶法 ¹ , 王樹卿 ¹

1.佳木斯大學附屬第一醫院普外一科（黑龍江佳木斯 154002）;;
2.青島大學附屬醫院甲狀腺外科（山東青島 266003）;

關鍵詞：

DNA甲基化凋亡增殖缺血再灌注小腸

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目的　探討缺血再灌注小腸中DNA甲基化對小腸細胞凋亡和增殖是否具有調控作用。
方法　將35只健康雄性Wistar大鼠隨機分為正常組（n=5）、假手術組（n=5）及缺血再灌注（0、3、6、12、24h，n=5）組。利用脫氧核糖核苷酸末端轉移酶介導的缺口末端標記測定法、透射電鏡和免疫組織化學方法分別檢測細胞凋亡和細胞增殖的變化，最后通過DNA甲基化位點組織末端酶鏈接檢測法檢測DNA甲基化。
結果　①與正常組和假手術組比較，缺血再灌注組缺血再灌注后3、6及12h時在小腸絨毛上皮、黏膜固有層及隱窩上皮中凋亡細胞均明顯增加（P＜0.01），且透射電鏡檢測證實，黏膜固有層的凋亡細胞主要是淋巴細胞和少量吞噬細胞。②與正常組和假手術組比較，缺血再灌注后3、6、12及24h時在小腸絨毛上皮中細胞增殖明顯（P＜0.01），缺血再灌注后6h和12h時在小腸黏膜固有層和隱窩上皮中細胞增殖明顯（P＜0.01）。③正常組和假手術組DNA甲基化在小腸絨毛上皮和隱窩上皮部分有弱表達；在缺血再灌注組中，小腸隱窩上皮部分DNA甲基化在近小腸干細胞部位表達最強，從隱窩上皮到絨毛上皮是以從強到弱的趨勢發生變化的。
結論　本研究的初步研究結果提示，缺血再灌注小腸中DNA甲基化可能對小腸細胞凋亡和增殖具有調控作用。

引用本文： 安樹才,王慶法,王樹卿. 缺血再灌注小腸中DNA甲基化對細胞凋亡和增殖的調控△. 中國普外基礎與臨床雜志, 2013, 20(11): 1234-1239. doi: 復制

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1. Hassoun HT， Zou L， Moore FA， et al. Alpha-melanocyte-stimulating hormone protects against mesenteric ischemia-reperfusion injury[J]. Am J Physiol Gastrointest Liver Physiol， 2002， 282(6)：G1059-G1068.
2. An S， Hishikawa Y， Koji T. Induction of cell death in rat small intestine by ischemia reperfusion：differential roles of Fas/Fas ligand and Bcl-2/Bax systems depending upon cell types[J]. Histochem Cell Biol， 2005， 123(3)：249-261.
3. Jackson-Grusby L， Beard C， Possemato R， et al. Loss of genomicmethylation causes p53-dependent apoptosis and epigenetic deregulation[J]. Nat Genet， 2001， 27(1)：31-39.
4. Herman JG， Baylin SB. Gene silencing in cancer in association with promoter hypermethylation[J]. N Engl J Med， 2003， 349(21)：2042-2054.
5. Rao X， Zhong J， Zhang S， et al. Loss of methyl-CpG-binding domain protein 2 enhances endothelial angiogenesis and protects mice against hind-limb ischemic injury[J]. Circulation， 2011， 123(25)：2964-2974.
6. Westberry JM， Wilson ME. Regulation of estrogen receptor alpha gene expression in the mouse prefrontal cortex during early postnatal development[J]. Neurogenetics， 2012， 13(2)：159-167.
7. Skowronski K， Dubey S， Rodenhiser D， et al. Ischemia dysregul-ates DNA methyltransferases and p16INK4a methylation in humancolorectal cancer cells[J]. Epigenetics， 2010， 5(6)：547-556.
8. Hashimoto S， Koji T， Niu J， et al. Differential staining of DNA strand breaks in dying cells by non-radioactive in situ nick transla-tion[J]. Arch Histol Cytol， 1995， 58(2)：161-170.
9. An S， Hishikawa Y， Liu J， et al．Lung injury after ischemia-reperfusion of small intestine in rats involves apoptosis of type Ⅱ alveolar epithelial cells mediated by TNF-alpha and activation of Bid pathway[J]. Apoptosis， 2007， 12(11)：1989-2001.
10. Koji T， Kondo S， Hishikawa Y， et al. In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites：a new principle of analysis of DNA methylation[J]. Histochem Cell Biol， 2008， 130(5)：917-925.
11. Otterson MF， Nie L， Schmidt JL， et al. EUK-207 protects humanintestinal microvascular endothelial cells (HIMEC) against irradi-ation-induced apoptosis through the Bcl2 pathway[J]. Life Sci， 2012， 91(15-16)：771-782.
12. Reardon C， Wang A， McKay DM. Transient local depletion of Foxp3+ regulatory T cells during recovery from colitis via Fas/Fas ligand-induced death[J]. J Immunol， 2008， 180(12)：8316-8326.
13. Neal MD， Sodhi CP， Jia H， et al. Toll-like receptor 4 is expressedon intestinal stem cells and regulates their proliferation and apoptosis via the p53 up-regulated modulator of apoptosis[J]. J Biol Chem， 2012， 287(44)：37296-37308.
14. Riehl TE， Foster L， Stenson WF. Hyaluronic acid is radioprote-ctive in the intestine through a TLR4 and COX-2-mediated mech-anism[J]. Am J Physiol Gastrointest Liver Physiol， 2012， 302(3)：G309-G316.
15. Booth C， Potten CS. Gut instincts：thoughts on intestinal epithelialstem cells[J]. J Clin Invest， 2000， 105(11)：1493-1499.
16. Dekaney CM， Fong JJ， Rigby RJ， et al. Expansion of intestinalstem cells associated with long-term adaptation following ileocecalresection in mice[J]. Am J Physiol Gastrointest Liver Physiol， 2007， 293(5)：1013-1022.
17. Shibata D. Mistakes make history：visualizing intestinal tectonics[J]. Gastroenterology， 2008， 134(2)：628-631.
18. McDonald SA， Greaves LC， Gutierrez-Gonzalez L， et al. Mech-anisms of field cancerization in the human stomach：the expansionand spread of mutated gastric stem cells[J]. Gastroenterology， 2008， 134(2)：500-510.
19. Barros SP， Offenbacher S. Epigenetics：connecting environment and genotype to phenotype and disease[J]. J Dent Res， 2009， 88(5)：400-408.
20. Dumitrescu RG. DNA methylation and histone modifications in breast cancer[J]. Methods Mol Biol， 2012， 863：35-45. doi：10.1007/978-1-61779-612-8_3.
21. Maegawa S， Hinkal G， Kim HS， et al. Widespread and tissuespecific age-related DNA methylation changes in mice[J]. Gen-ome Res， 2010， 20(3)：332-340.
22. Kocemba K， Groen R， van Andel H， et al. Transcriptional silen-cing of the Wnt-Antagonist DKK1 by promoter methylation isassociated with enhanced Wnt signaling in advanced multiple mye-loma[J]. PLoS One， 2012， 7：e30359.
23. Hur K， Niwa T， Toyoda T， et al. Insufficient role of cell proliferation in aberrant DNA methylation induction and involvement of specific types of inflammation[J]. Carcinogenesis， 2011， 32(1)：35-41.
24. Eslaminejad MB， Fani N， Shahhoseini M. Epigenetic regulation of osteogenic and chondrogenic differentiation of mesenchymal stem cells in culture[J]. Cell， 2013， 15(1)：1-10．.
25. 李望，錢國偉，徐為，等. DNA甲基化對Ki-67啟動子轉錄活性的影響[J]. 中華實驗外科雜志， 2011， 28(6)：901-903.

《中國普外基礎與臨床雜志》

缺血再灌注小腸中DNA甲基化對細胞凋亡和增殖的調控△

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《中國普外基礎與臨床雜志》

缺血再灌注小腸中DNA甲基化對細胞凋亡和增殖的調控△

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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