中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
2期
251-258
,共8页
组织构建%组织工程%角膜基质细胞%组织因子途径抑制物2%基质金属蛋白酶%细胞外基质%角膜新生血管%国家自然科学基金
組織構建%組織工程%角膜基質細胞%組織因子途徑抑製物2%基質金屬蛋白酶%細胞外基質%角膜新生血管%國傢自然科學基金
조직구건%조직공정%각막기질세포%조직인자도경억제물2%기질금속단백매%세포외기질%각막신생혈관%국가자연과학기금
corneal stroma%metalloproteinase%extracellular matrix%cornea%culture medium
背景：有研究表明，基质金属蛋白酶所参与的细胞外基质降解在角膜新生血管形成过程中起关键作用，组织因子途径抑制物2是新近发现的一种新型的丝氨酸蛋白酶抑制物，能有效抑制基质金属蛋白酶的活性。<br> 目的：观察组织因子途径抑制物2对体外角膜基质细胞表达基质金属蛋白酶活性的关系。<br> 方法：在体外对兔角膜基质细胞进行原代及传代培养，用脂质体介导的人类组织因子途径抑制物2真核表达载体转染兔角膜基质细胞，G418筛选阳性细胞。<br> 结果与结论：RT-PCR，Western blot及明胶酶谱法分析结果显示，转染后角膜基质细胞组织因子途径抑制物2 mRNA和蛋白质的表达均上调(P<0.05)，而基质金属蛋白酶1，2的活性下降(P<0.05)。结果提示，组织因子途径抑制物2可明显抑制角膜基质细胞中基质金属蛋白酶1，2的活性。
揹景：有研究錶明，基質金屬蛋白酶所參與的細胞外基質降解在角膜新生血管形成過程中起關鍵作用，組織因子途徑抑製物2是新近髮現的一種新型的絲氨痠蛋白酶抑製物，能有效抑製基質金屬蛋白酶的活性。<br> 目的：觀察組織因子途徑抑製物2對體外角膜基質細胞錶達基質金屬蛋白酶活性的關繫。<br> 方法：在體外對兔角膜基質細胞進行原代及傳代培養，用脂質體介導的人類組織因子途徑抑製物2真覈錶達載體轉染兔角膜基質細胞，G418篩選暘性細胞。<br> 結果與結論：RT-PCR，Western blot及明膠酶譜法分析結果顯示，轉染後角膜基質細胞組織因子途徑抑製物2 mRNA和蛋白質的錶達均上調(P<0.05)，而基質金屬蛋白酶1，2的活性下降(P<0.05)。結果提示，組織因子途徑抑製物2可明顯抑製角膜基質細胞中基質金屬蛋白酶1，2的活性。
배경：유연구표명，기질금속단백매소삼여적세포외기질강해재각막신생혈관형성과정중기관건작용，조직인자도경억제물2시신근발현적일충신형적사안산단백매억제물，능유효억제기질금속단백매적활성。<br> 목적：관찰조직인자도경억제물2대체외각막기질세포표체기질금속단백매활성적관계。<br> 방법：재체외대토각막기질세포진행원대급전대배양，용지질체개도적인류조직인자도경억제물2진핵표체재체전염토각막기질세포，G418사선양성세포。<br> 결과여결론：RT-PCR，Western blot급명효매보법분석결과현시，전염후각막기질세포조직인자도경억제물2 mRNA화단백질적표체균상조(P<0.05)，이기질금속단백매1，2적활성하강(P<0.05)。결과제시，조직인자도경억제물2가명현억제각막기질세포중기질금속단백매1，2적활성。
BACKGROUND:The degradation of extracellular matrix, which is mediated by matrix metal oproteinases, plays a crucial role in the corneal neovascularization. Tissue factor pathway inhibitor 2, a new type serine proteinase inhibitor, can effectively inhibit the activity of matrix metal oproteinases. <br> OBJECTIVE:To elucidate the effect of tissue factor pathway inhibitor 2 on the expressions of matrix metal oproteinases in keratocytes in vitro. <br> METHODS:Rabbit keratocytes were primarily cultured and subcultured in vitro. Plasmid vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes with Lipofectamine 2000. The positive cells were selected using G418. <br> RESULTS AND CONCLUSION:The results of reverse transcription-polymerase chain reaction, western blot analysis and gelatinase zymography analysis showed that, expression of mRNA and protein of tissue factor pathway inhibitor 2 was upregulated in the transfected keratocytes (P<0.05), while activity of matrix metal oproteinase 1 and 2 was significantly decreased (P<0.05). Experimental findings indicate that that tissue factor pathway inhibitor 2 strongly inhibits the activity of matrix metal oproteinase 1 and 2 in keratocytes.
