熱損傷角質形成細胞培養上清對真皮成纖維細胞Ⅰ、Ⅲ型膠原及基質金屬蛋白酶1表達的影響_《中國修復重建外科雜志》

作者：

楊薛康 , 白曉智 , 張月 , 李小強 , 石繼紅 , 湯朝武 , 胡大海

第四軍醫大學西京醫院全軍燒傷中心（西安，710032） △為共同第一作者;

關鍵詞：

角質形成細胞真皮成纖維細胞熱損傷 Ⅰ型膠原 Ⅲ型膠原基質金屬蛋白酶1

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目的探討角質形成細胞（keratinocytes，KC）熱損傷后對真皮成纖維細胞（fibroblasts，Fb）Ⅰ、Ⅲ型膠原及基質金屬蛋白酶1（matrix metalloproteinase 1，MMP-1）表達的影響。方法分離培養人正常Fb及KC，分別建立KC、Fb熱損傷模型；收集正常及熱損傷12 h后細胞培養上清，并配制成濃度為50%的細胞條件培養液。根據培養液不同將第3～5代Fb分為3組，分別采用含50%熱損傷KC培養上清（A組）、含50%正常KC培養上清（B組）的條件培養液及單純DMEM（C組）培養，24 h后收集3組細胞；另于培養0、1、2、6、12、24、48 h分別收集A組細胞。采用含50%熱損傷Fb培養上清的條件培養液培養Fb，于0、1、2、6、12、24、48 h收集細胞。采用實時熒光定量PCR檢測各時間點KC熱損傷條件培養上清對 Fb Ⅰ、Ⅲ型膠原及MMP-1 mRNA表達影響，以及Fb熱損傷條件培養上清對Fb MMP-1 mRNA表達影響。結果KC熱損傷條件培養上清培養24 h，A組Ⅰ、Ⅲ型膠原及MMP-1 mRNA相對表達量均顯著高于B、C組，差異有統計學意義（P lt; 0.05）。培養2、6、12、24、48 h，A組Ⅰ、Ⅲ型膠原及MMP-1 mRNA相對表達量均高于0 h（P lt; 0.05），1 h與0 h差異無統計學意義（P gt; 0.05）；隨培養時間延長相對表達量逐漸增高，2 h后各時間點間比較差異均有統計學意義（P lt; 0.05）。Fb熱損傷條件培養上清培養1、2、6、12、24、48 h，MMP-1 mRNA相對表達量與0 h比較，差異均有統計學意義（P lt; 0.05）；培養2 h后相對表達量逐漸降低，各時間點間差異均有統計學意義（P lt; 0.05）。結論熱損傷后KC培養上清對Fb Ⅰ、Ⅲ型膠原及MMP-1的表達具有調控作用。

引用本文： 楊薛康,白曉智,張月,李小強,石繼紅,湯朝武,胡大海. 熱損傷角質形成細胞培養上清對真皮成纖維細胞Ⅰ、Ⅲ型膠原及基質金屬蛋白酶1表達的影響. 中國修復重建外科雜志, 2012, 26(12): 1497-1500. doi: 復制

1.	沈丹蓓, 夏隆慶. 角質形成細胞與皮膚創面愈合瘢痕形成. 中華整形外科雜志, 2005, 21(1): 69-71.
2.	白曉智, 呂根法, 謝松濤, 等. 離體角質形成細胞熱損傷模型建立及細胞凋亡觀察. 中華燒傷雜志, 2009, 25(3): 189-192.
3.	Xia W, Phan TT, Lim IJ, et al. Complex epithelial-mesenchymal interactions modulate transforming growth factor-beta expression in keloid-derived cells. Wound Repair Regen, 2004, 12(5): 546-556.
4.	El Ghalbzouri A, Hensbergen P, Gibbs S, et al. Fibroblasts facilitate re-epithelialization in wounded human skin equivalents. Lab Invest, 2004, 84(1): 102-112.
5.	Werner S, Krieg T, Smola H. Keratinocyte-fibroblast interactions in wound healing. J Invest Dermatol, 2007, 127(5): 998-1008.
6.	van der Veer WM, Bloemen MC, Ulrich MM, et al. Potential cellular and molecular causes of hypertrophic scar formation. Burns, 2009, 35(1): 15-29.
7.	Wang X, Liu Y, Deng Z, et al. Inhibition of dermal fibrosis in self-assembled skin equivalents by undifferentiated keratinocytes. J Dermatol Sci, 2009, 53(2): 103-111.
8.	Bellemare J, Roberge CJ, Bergeron D, et al. Epidermis promotes dermal fibrosis: role in the pathogenesis of hypertrophic scars. J Pathol, 2005, 206(1): 1-8.
9.	Harrison CA, Gossiel F, Bullock AJ, et al. Investigation of keratinocyte regulation of collagen I synthesis by dermal fibroblasts in a simple in vitro model. Br J Dermatol, 2006, 154(3): 401-410.
10.	Tandara AA, Kloeters O, Mogford JE, et al. Hydrated keratinocytes reduce collagen synthesis by fibroblasts via paracrine mechanisms. Wound Repair Regen, 2007, 15(4): 497-504.
11.	馬忠鋒, 柴家科. 表皮細胞的培養移植的現狀與展望. 中華燒傷雜志, 2005, 21(2): 158-160.
12.	Vashishta A, Saraswat Ohri S, Vetvickova J, et al. Procathepsin D secreted by HaCaT keratinocyte cells—A novel regulator of keratinocyte growth. Eur J Cell Biol, 2007, 86(6): 303-313.
13.	白曉智, 胡大海, 石繼紅, 等. 熱損傷角質形成細胞培養上清對真皮成纖維細胞增殖與膠原mRNA表達的影響. 中華實驗外科雜志, 2011, 28(3): 380-382.
14.	白曉智, 胡大海, 張萬福, 等. 熱損傷角質形成細胞培養上清液對成纖維細胞生物學行為的影響. 中華燒傷雜志, 2010, 26(2): 133-137.
15.	Moulin V, Larochelle S, Langlois C, et al. Normal skin wound and hypertrophic scar myofibroblasts have differential responses to apoptotic inductors. J Cell Physiol, 2004, 198(3): 350-358.
16.	陳蘇麗, 劉流. 上皮細胞-干細胞在創傷愈合中的作用. 中國組織工程研究與臨床康復, 2010, 14(6): 1092-1096.
17.	Kisseleva T, Brenner DA. Mechanisms of fibrogenesis. Exp Biol Med, 2008, 233(2): 109-122.
18.	Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology, 2008, 134(6): 1655-1669．.
19.	Risinger GM Jr, Hunt TS, Updike DL, et al. Matrix metalloproteinase-2 expression by vascular smooth muscle cells is mediated by both stimulatory and inhibitory signals in response to growth factors. J Biol Chem, 2006, 281(36): 25915-25925.
20.	Ravanti L, Kähäri VM. Matrix metalloproteinases in wound repair (review). Int J Mol Med, 2000, 6(4): 391-407.
21.	Longaker MT, Whitby DJ, Adzick NS, et al. Studies in fetal wound healing, VI. Second and early third trimester fetal wounds demonstrate rapid collagen deposition without scar formation. J Pediatr Surg, 1990, 25(1): 63-69.

1. 沈丹蓓, 夏隆慶. 角質形成細胞與皮膚創面愈合瘢痕形成. 中華整形外科雜志, 2005, 21(1): 69-71.
2. 白曉智, 呂根法, 謝松濤, 等. 離體角質形成細胞熱損傷模型建立及細胞凋亡觀察. 中華燒傷雜志, 2009, 25(3): 189-192.
3. Xia W, Phan TT, Lim IJ, et al. Complex epithelial-mesenchymal interactions modulate transforming growth factor-beta expression in keloid-derived cells. Wound Repair Regen, 2004, 12(5): 546-556.
4. El Ghalbzouri A, Hensbergen P, Gibbs S, et al. Fibroblasts facilitate re-epithelialization in wounded human skin equivalents. Lab Invest, 2004, 84(1): 102-112.
5. Werner S, Krieg T, Smola H. Keratinocyte-fibroblast interactions in wound healing. J Invest Dermatol, 2007, 127(5): 998-1008.
6. van der Veer WM, Bloemen MC, Ulrich MM, et al. Potential cellular and molecular causes of hypertrophic scar formation. Burns, 2009, 35(1): 15-29.
7. Wang X, Liu Y, Deng Z, et al. Inhibition of dermal fibrosis in self-assembled skin equivalents by undifferentiated keratinocytes. J Dermatol Sci, 2009, 53(2): 103-111.
8. Bellemare J, Roberge CJ, Bergeron D, et al. Epidermis promotes dermal fibrosis: role in the pathogenesis of hypertrophic scars. J Pathol, 2005, 206(1): 1-8.
9. Harrison CA, Gossiel F, Bullock AJ, et al. Investigation of keratinocyte regulation of collagen I synthesis by dermal fibroblasts in a simple in vitro model. Br J Dermatol, 2006, 154(3): 401-410.
10. Tandara AA, Kloeters O, Mogford JE, et al. Hydrated keratinocytes reduce collagen synthesis by fibroblasts via paracrine mechanisms. Wound Repair Regen, 2007, 15(4): 497-504.
11. 馬忠鋒, 柴家科. 表皮細胞的培養移植的現狀與展望. 中華燒傷雜志, 2005, 21(2): 158-160.
12. Vashishta A, Saraswat Ohri S, Vetvickova J, et al. Procathepsin D secreted by HaCaT keratinocyte cells—A novel regulator of keratinocyte growth. Eur J Cell Biol, 2007, 86(6): 303-313.
13. 白曉智, 胡大海, 石繼紅, 等. 熱損傷角質形成細胞培養上清對真皮成纖維細胞增殖與膠原mRNA表達的影響. 中華實驗外科雜志, 2011, 28(3): 380-382.
14. 白曉智, 胡大海, 張萬福, 等. 熱損傷角質形成細胞培養上清液對成纖維細胞生物學行為的影響. 中華燒傷雜志, 2010, 26(2): 133-137.
15. Moulin V, Larochelle S, Langlois C, et al. Normal skin wound and hypertrophic scar myofibroblasts have differential responses to apoptotic inductors. J Cell Physiol, 2004, 198(3): 350-358.
16. 陳蘇麗, 劉流. 上皮細胞-干細胞在創傷愈合中的作用. 中國組織工程研究與臨床康復, 2010, 14(6): 1092-1096.
17. Kisseleva T, Brenner DA. Mechanisms of fibrogenesis. Exp Biol Med, 2008, 233(2): 109-122.
18. Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology, 2008, 134(6): 1655-1669．.
19. Risinger GM Jr, Hunt TS, Updike DL, et al. Matrix metalloproteinase-2 expression by vascular smooth muscle cells is mediated by both stimulatory and inhibitory signals in response to growth factors. J Biol Chem, 2006, 281(36): 25915-25925.
20. Ravanti L, Kähäri VM. Matrix metalloproteinases in wound repair (review). Int J Mol Med, 2000, 6(4): 391-407.
21. Longaker MT, Whitby DJ, Adzick NS, et al. Studies in fetal wound healing, VI. Second and early third trimester fetal wounds demonstrate rapid collagen deposition without scar formation. J Pediatr Surg, 1990, 25(1): 63-69.

《中國修復重建外科雜志》

熱損傷角質形成細胞培養上清對真皮成纖維細胞Ⅰ、Ⅲ型膠原及基質金屬蛋白酶1表達的影響

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

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《中國修復重建外科雜志》

熱損傷角質形成細胞培養上清對真皮成纖維細胞Ⅰ、Ⅲ型膠原及基質金屬蛋白酶1表達的影響

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

摘要全文圖表視頻參考文獻施引文獻補充材料