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

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(diào)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(zhuǎn)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(jié)果 ?、倥c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(fā)生變化的。
結(jié)論 　本研究的初步研究結(jié)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(diào)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(diào)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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 15. Booth C， Potten CS. Gut instincts：thoughts on intestinal epithelialstem cells[J]. J Clin Invest， 2000， 105(11)：1493-1499.
  16. 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. 17. Shibata D. Mistakes make history：visualizing intestinal tectonics[J]. Gastroenterology， 2008， 134(2)：628-631.
  18. 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. 19. Barros SP， Offenbacher S. Epigenetics：connecting environment and genotype to phenotype and disease[J]. J Dent Res， 2009， 88(5)：400-408.
  20. 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. 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. 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. 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. 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. 25. 李望， 錢國偉， 徐為， 等. DNA甲基化對Ki-67啟動子轉(zhuǎn)錄活性的影響[J]. 中華實驗外科雜志， 2011， 28(6)：901-903.