目的探討角質(zhì)形成細(xì)胞（keratinocytes，KC）熱損傷后對(duì)真皮成纖維細(xì)胞（fibroblasts，F(xiàn)b）Ⅰ、Ⅲ型膠原及基質(zhì)金屬蛋白酶1（matrix metalloproteinase 1，MMP-1）表達(dá)的影響。 方法分離培養(yǎng)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(xì)胞培養(yǎng)上清，并配制成濃度為50%的細(xì)胞條件培養(yǎng)液。根據(jù)培養(yǎng)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(yǎng)上清（A組）、含50%正常KC培養(yǎng)上清（B組）的條件培養(yǎng)液及單純DMEM（C組）培養(yǎng)，24 h后收集3組細(xì)胞；另于培養(yǎng)0、1、2、6、12、24、48 h分別收集A組細(xì)胞。采用含50%熱損傷Fb培養(yǎng)上清的條件培養(yǎng)液培養(yǎng)Fb，于0、1、2、6、12、24、48 h收集細(xì)胞。采用實(shí)時(shí)熒光定量PCR檢測(cè)各時(shí)間點(diǎn)KC熱損傷條件培養(yǎng)上清對(duì) Fb Ⅰ、Ⅲ型膠原及MMP-1 mRNA表達(dá)影響，以及Fb熱損傷條件培養(yǎng)上清對(duì)Fb MMP-1 mRNA表達(dá)影響。 結(jié)果KC熱損傷條件培養(yǎng)上清培養(yǎng)24 h，A組Ⅰ、Ⅲ型膠原及MMP-1 mRNA相對(duì)表達(dá)量均顯著高于B、C組，差異有統(tǒng)計(jì)學(xué)意義（P lt; 0.05）。培養(yǎng)2、6、12、24、48 h，A組Ⅰ、Ⅲ型膠原及MMP-1 mRNA相對(duì)表達(dá)量均高于0 h（P lt; 0.05），1 h與0 h差異無統(tǒng)計(jì)學(xué)意義（P gt; 0.05）；隨培養(yǎng)時(shí)間延長相對(duì)表達(dá)量逐漸增高，2 h后各時(shí)間點(diǎn)間比較差異均有統(tǒng)計(jì)學(xué)意義（P lt; 0.05）。Fb熱損傷條件培養(yǎng)上清培養(yǎng)1、2、6、12、24、48 h，MMP-1 mRNA相對(duì)表達(dá)量與0 h比較，差異均有統(tǒng)計(jì)學(xué)意義（P lt; 0.05）；培養(yǎng)2 h后相對(duì)表達(dá)量逐漸降低，各時(shí)間點(diǎn)間差異均有統(tǒng)計(jì)學(xué)意義（P lt; 0.05）。 結(jié)論熱損傷后KC培養(yǎng)上清對(duì)Fb Ⅰ、Ⅲ型膠原及MMP-1的表達(dá)具有調(diào)控作用。
引用本文： 楊薛康,白曉智,張?jiān)?李小強(qiáng),石繼紅,湯朝武,胡大海. 熱損傷角質(zhì)形成細(xì)胞培養(yǎng)上清對(duì)真皮成纖維細(xì)胞Ⅰ、Ⅲ型膠原及基質(zhì)金屬蛋白酶1表達(dá)的影響. 中國修復(fù)重建外科雜志, 2012, 26(12): 1497-1500. doi: 復(fù)制
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| 2. | 白曉智, 呂根法, 謝松濤, 等. 離體角質(zhì)形成細(xì)胞熱損傷模型建立及細(xì)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(xì)胞的培養(yǎng)移植的現(xiàn)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(zhì)形成細(xì)胞培養(yǎng)上清對(duì)真皮成纖維細(xì)胞增殖與膠原mRNA表達(dá)的影響. 中華實(shí)驗(yàn)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(zhì)形成細(xì)胞培養(yǎng)上清液對(duì)成纖維細(xì)胞生物學(xué)行為的影響. 中華燒傷雜志, 2010, 26(2): 133-137. |
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| 16. | 陳蘇麗, 劉流. 上皮細(xì)胞-干細(xì)胞在創(chuàng)傷愈合中的作用. 中國組織工程研究與臨床康復(fù), 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(zhì)形成細(xì)胞與皮膚創(chuàng)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(zhì)形成細(xì)胞熱損傷模型建立及細(xì)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(xì)胞的培養(yǎng)移植的現(xiàn)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(zhì)形成細(xì)胞培養(yǎng)上清對(duì)真皮成纖維細(xì)胞增殖與膠原mRNA表達(dá)的影響. 中華實(shí)驗(yàn)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(zhì)形成細(xì)胞培養(yǎng)上清液對(duì)成纖維細(xì)胞生物學(xué)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(xì)胞-干細(xì)胞在創(chuàng)傷愈合中的作用. 中國組織工程研究與臨床康復(fù), 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.