中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2015年
2期
89-94
,共6页
扁平苔藓,口腔%缺氧诱导因子1,α亚基%基质金属蛋白酶9%角质形成细胞
扁平苔蘚,口腔%缺氧誘導因子1,α亞基%基質金屬蛋白酶9%角質形成細胞
편평태선,구강%결양유도인자1,α아기%기질금속단백매9%각질형성세포
Lichen planus,oral%Hypoxia-inducible factor 1,alpha subunit%Matrix metalloproteinase 9%Keratinocytes
目的 探讨缺氧对人口腔扁平苔藓(oral lichen planus,OLP)角质形成细胞增殖及缺氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1 α)、血管内皮生长因子(vascular endothelial growth factor,VEGF)及基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)表达的影响,以期为OLP发病机制的研究提供新思路.方法 体外分离培养OLP角质形成细胞;使用密闭培养箱模拟缺氧环境,分为常氧组和缺氧组,两组细胞分别培养12、24、36及48 h,采用细胞计数试剂盒8的方法监测缺氧各时点细胞增殖状况,并确定进一步实验干预时间点,每组实验均重复3次;SYBR Green Ⅰ荧光染料实时定量PCR法和蛋白质印迹法分别检测各组细胞HIF-1α、VEGF、MMP-9 mRNA和蛋白表达水平,并采用相关性分析研究VEGF、MMP-9与HIF-1α的相关性.结果 24、36、48 h缺氧组OLP角质形成细胞增殖力(A值)(分别为0.340±0.002、0.415±0.006、0.546±0.006)均显著低于同时间点常氧组(分别为0.431 ±0.001、0.620±0.004、1.022±0.005),P＜0.01; 24、36、48 h缺氧组VEGF(2.087±0.291、3.189±0.573、5.402±0.563)及MMP-9(2.936±0.500、4.083±0.300、6.374±0.858)的mRNA表达量均显著高于常氧组(P＜0.05),HIF-1α (0.414±0.093、0.751±0.056、0.875±0.040)、VEGF (0.393±0.046、0.557±0.078、0.767±0.045)及MMP-9 (0.250±0.053、0.384±0.038、0.611 ±0.092)的蛋白表达量均显著高于常氧组(P＜0.05).HIF-1α与VEGF(r =0.905,P=0.000)及MMP-9(r =0.881,P=0.000)的蛋白表达水平均呈显著正相关.结论 缺氧在一定时间内可抑制OLP角质形成细胞增殖,上调细胞中HIF-1α蛋白、VEGF及MMP-9 mRNA和蛋白的表达;缺氧环境下HIF-1α可能通过调控下游靶基因参与OLP的病情进展.
目的 探討缺氧對人口腔扁平苔蘚(oral lichen planus,OLP)角質形成細胞增殖及缺氧誘導因子1α(hypoxia-inducible factor-1α,HIF-1 α)、血管內皮生長因子(vascular endothelial growth factor,VEGF)及基質金屬蛋白酶9(matrix metalloproteinase-9,MMP-9)錶達的影響,以期為OLP髮病機製的研究提供新思路.方法 體外分離培養OLP角質形成細胞;使用密閉培養箱模擬缺氧環境,分為常氧組和缺氧組,兩組細胞分彆培養12、24、36及48 h,採用細胞計數試劑盒8的方法鑑測缺氧各時點細胞增殖狀況,併確定進一步實驗榦預時間點,每組實驗均重複3次;SYBR Green Ⅰ熒光染料實時定量PCR法和蛋白質印跡法分彆檢測各組細胞HIF-1α、VEGF、MMP-9 mRNA和蛋白錶達水平,併採用相關性分析研究VEGF、MMP-9與HIF-1α的相關性.結果 24、36、48 h缺氧組OLP角質形成細胞增殖力(A值)(分彆為0.340±0.002、0.415±0.006、0.546±0.006)均顯著低于同時間點常氧組(分彆為0.431 ±0.001、0.620±0.004、1.022±0.005),P＜0.01; 24、36、48 h缺氧組VEGF(2.087±0.291、3.189±0.573、5.402±0.563)及MMP-9(2.936±0.500、4.083±0.300、6.374±0.858)的mRNA錶達量均顯著高于常氧組(P＜0.05),HIF-1α (0.414±0.093、0.751±0.056、0.875±0.040)、VEGF (0.393±0.046、0.557±0.078、0.767±0.045)及MMP-9 (0.250±0.053、0.384±0.038、0.611 ±0.092)的蛋白錶達量均顯著高于常氧組(P＜0.05).HIF-1α與VEGF(r =0.905,P=0.000)及MMP-9(r =0.881,P=0.000)的蛋白錶達水平均呈顯著正相關.結論 缺氧在一定時間內可抑製OLP角質形成細胞增殖,上調細胞中HIF-1α蛋白、VEGF及MMP-9 mRNA和蛋白的錶達;缺氧環境下HIF-1α可能通過調控下遊靶基因參與OLP的病情進展.
목적 탐토결양대인구강편평태선(oral lichen planus,OLP)각질형성세포증식급결양유도인자1α(hypoxia-inducible factor-1α,HIF-1 α)、혈관내피생장인자(vascular endothelial growth factor,VEGF)급기질금속단백매9(matrix metalloproteinase-9,MMP-9)표체적영향,이기위OLP발병궤제적연구제공신사로.방법 체외분리배양OLP각질형성세포;사용밀폐배양상모의결양배경,분위상양조화결양조,량조세포분별배양12、24、36급48 h,채용세포계수시제합8적방법감측결양각시점세포증식상황,병학정진일보실험간예시간점,매조실험균중복3차;SYBR Green Ⅰ형광염료실시정량PCR법화단백질인적법분별검측각조세포HIF-1α、VEGF、MMP-9 mRNA화단백표체수평,병채용상관성분석연구VEGF、MMP-9여HIF-1α적상관성.결과 24、36、48 h결양조OLP각질형성세포증식력(A치)(분별위0.340±0.002、0.415±0.006、0.546±0.006)균현저저우동시간점상양조(분별위0.431 ±0.001、0.620±0.004、1.022±0.005),P＜0.01; 24、36、48 h결양조VEGF(2.087±0.291、3.189±0.573、5.402±0.563)급MMP-9(2.936±0.500、4.083±0.300、6.374±0.858)적mRNA표체량균현저고우상양조(P＜0.05),HIF-1α (0.414±0.093、0.751±0.056、0.875±0.040)、VEGF (0.393±0.046、0.557±0.078、0.767±0.045)급MMP-9 (0.250±0.053、0.384±0.038、0.611 ±0.092)적단백표체량균현저고우상양조(P＜0.05).HIF-1α여VEGF(r =0.905,P=0.000)급MMP-9(r =0.881,P=0.000)적단백표체수평균정현저정상관.결론 결양재일정시간내가억제OLP각질형성세포증식,상조세포중HIF-1α단백、VEGF급MMP-9 mRNA화단백적표체;결양배경하HIF-1α가능통과조공하유파기인삼여OLP적병정진전.
Objective To investigate the effect of hypoxia on the proliferation and expressions of hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9(MMP-9) in keratinocytes obtained from oral lichen planus(OLP) lesions.Methods Hypoxia environment was induced by a airtight incubator.Five groups were included,normoxia control group,hypoxia control group(12,24,36,48 h).The effect of different treatment time of hypoxia on cellular proliferation was determined with cell counting kit-8(CCK-8).The mRNA and protein expressions of HIF-lα,VEGF and MMP-9 were analyzed respectively by quantitative real-time polymerase chain reaction with SYBR Green Ⅰ and Western blotting.Results The growth activity of keratinocytes obtained from OLP lesions in the hypoxia groups(0.340±0.002,0.415±0.006,0.546±0.006) was reduced than that in control (0.431±0.001,0.620±0.004,1.022±0.005)(P＜0.01).The mRNA levels of VEGF(2.087±0.291,3.189±0.573,5.402±0.563) and MMP-9(2.936±0.500,4.083±0.300,6.374±0.858) were elevated by hypoxia(P＜0.05).The protein levels of HIF-lα(0.414±0.093,0.751 ±0.056,0.875±0.040),VEGF(0.393 ±0.046,0.557±0.078,0.767±0.045) and MMP-9(0.250±0.053,0.384±0.038,0.611±0.092) were all remarkably elevated by hypoxia (P＜O.05).However,hypoxia had no effect on HIF-1α mRNA expression.The mRNA expression of HIF-1α after hypoxia exposure for 36 h(1.412±0.094) and 48 h(1.417±0.446) was higher than that of control group,however,there was no significant difference.A positive correlation was noted between HIF-1α and VEGF in protein level(r=0.905,P=O.O00),and the same correlation found between HIF-1α and MMP-9(r=0.881,P=0.000).Conclusions Hypoxia conditions may inhibit the proliferation of keratinocytes obtained from OLP lesions.Hypoxia conditions can promote the protein expressions of HIF-1α and both the mRNA and protein expression of VEGF and MMP-9 in keratinocytes obtained from OLP lesions exposed to hypoxia.The relative high expression of HIF-lα may be involved in multiple aspects of OLP progression through the regulation of its downstream target genes.
