CAJ | 학술논문

目的观察花生四烯酸对人增生性瘢痕(HS)成纤维细胞体外增殖的影响.方法体外培养人HS纤维细胞,取第4代细胞用于实验.分别用0、5、10、20、40、80 μmol/L花生四烯酸作用于HS成纤维细胞24、48、72 h后,采用细胞计数试剂盒检测HS成纤维细胞增殖活力.制作HS成纤维细胞爬片,免疫细胞化学法检测上述不同浓度花生四烯酸作用24 h后细胞核中Ki-67抗原的表达情况.结果 HS成纤维细胞在未加花生四烯酸干预时,生长旺盛,形态规则,胞质丰富,胞核呈椭圆或圆形,细胞生长走向基本一致;花生四烯酸浓度为5、10 μmol/L时,细胞生长形态与未加花生四烯酸干预时未见明显区别;花生四烯酸浓度为20、40、80μmol/L时,HS成纤维细胞与未加花生四烯酸干预时比较,数目减少,细胞形状改变,部分细胞突出回缩,呈圆钝形,胞核浓缩,培养液内漂浮细胞较多.花生四烯酸作用于HS成纤维细胞24 h后,5、10、20、40、80 μmol/L组增殖抑制率分别为(1.2±1.5)％、(2.9±2.2)％、(8.8±2.3)％、(22.0±2.3)％、(37.9±2.1)％,48 h后分别为(4.3±1.4)％、(8.7±1.8)％、(18.3±1.5)％、(27.6±0.8)％、(42.7±2.2)％,72 h后分别为(4.7±1.7)％、(9.6±2.0)％、(21.6±2.1)％、(30.3±2.1)％、(47.0±2.6)％,在20 ～ 80μmol/L花生四烯酸浓度范围内,HS成纤维细胞生长抑制率随着花生四烯酸作用浓度和时间的增加而增加(P＜0.05).花生四烯酸浓度为5、10 μmol/L时,Ki-67抗原表达阳性率与未加花生四烯酸时比较,差异无统计学意义(P＞0.05);花生四烯酸浓度为20、40、80 μmol/L时,Ki-67抗原表达阳性率均明显低于未加花生四烯酸时[(47.1±1.6)％、(32.1±4.4)％、(18.6±2.3)％比(58.7±4.6)％](P＜0.05).结论 20 ～ 80 μmol/L的花生四烯酸能抑制人HS成纤维细胞的增殖,可能是其抑制瘢痕增生作用机制之一.
목적 관찰화생사희산대인증생성반흔(HS)성섬유세포체외증식적영향.방법 체외배양인HS섬유세포,취제4대세포용우실험.분별용0、5、10、20、40、80 μmol/L화생사희산작용우HS성섬유세포24、48、72 h후,채용세포계수시제합검측HS성섬유세포증식활력.제작HS성섬유세포파편,면역세포화학법검측상술불동농도화생사희산작용24 h후세포핵중Ki-67항원적표체정황.결과 HS성섬유세포재미가화생사희산간예시,생장왕성,형태규칙,포질봉부,포핵정타원혹원형,세포생장주향기본일치;화생사희산농도위5、10 μmol/L시,세포생장형태여미가화생사희산간예시미견명현구별;화생사희산농도위20、40、80μmol/L시,HS성섬유세포여미가화생사희산간예시비교,수목감소,세포형상개변,부분세포돌출회축,정원둔형,포핵농축,배양액내표부세포교다.화생사희산작용우HS성섬유세포24 h후,5、10、20、40、80 μmol/L조증식억제솔분별위(1.2±1.5)％、(2.9±2.2)％、(8.8±2.3)％、(22.0±2.3)％、(37.9±2.1)％,48 h후분별위(4.3±1.4)％、(8.7±1.8)％、(18.3±1.5)％、(27.6±0.8)％、(42.7±2.2)％,72 h후분별위(4.7±1.7)％、(9.6±2.0)％、(21.6±2.1)％、(30.3±2.1)％、(47.0±2.6)％,재20 ～ 80μmol/L화생사희산농도범위내,HS성섬유세포생장억제솔수착화생사희산작용농도화시간적증가이증가(P＜0.05).화생사희산농도위5、10 μmol/L시,Ki-67항원표체양성솔여미가화생사희산시비교,차이무통계학의의(P＞0.05);화생사희산농도위20、40、80 μmol/L시,Ki-67항원표체양성솔균명현저우미가화생사희산시[(47.1±1.6)％、(32.1±4.4)％、(18.6±2.3)％비(58.7±4.6)％](P＜0.05).결론 20 ～ 80 μmol/L적화생사희산능억제인HS성섬유세포적증식,가능시기억제반흔증생작용궤제지일.
Objective To observe the effects of arachidonic acid on proliferation of human hypertrophic scar fibroblasts in vitro.Methods The human hypertrophic scar fibroblasts were cultured in vitro and the fourth generation cells were used in the experiment.The cultured fibroblasts were treated with arachidonic acid (0, 5,10, 20, 40, 80 μmol/L) for 24, 48 and 72 h, then the proliferation of cells was measured by cell counting kit,the expression of Ki-67 protein was detected by immunocytochemical method.Results The normal hypertrophic scar fibroblasts grew well with regular morphology, the cytoplasm was rich, the nucleus was oval or round, the cell shape was basically the same;after treated with arachidonic acid of 5 μmol/L and 10 μmol/L, the cell morphology was not significantly changed;after treated with arachidonic acid of 20, 40 and 80 μmol/L, the cell shape was irregular, the cell number was reduced, the cell size was not uniform, the synapses of partial cells were retracted and become blunt round, the nucleus was condensed and many floating cells were observed.The inhibitory rate of proliferation was (1.2±1.5)％, (2.9 ±2.2)％, (8.8 ±2.3)％, (22.0±2.3)％, (37.9 ±2.1)％ after treated with 5, 10, 20, 40, 80 μmol/L of arachidonic acid for 24 h, (4.3 ± 1.4) ％, (8.7 ± 1.8) ％ , (18.3 ±1.5)％, (27.6±0.8)％, (42.7 ±2.2)％ for48 h, (4.7 ± 1.7)％, (9.6 ±2.0)％, (21.6 ±2.1)％,(30.3 ± 2.1) ％ , (47.0 ± 2.6) ％ for 72 h.In 20-80 μmol/L arachidonic acid concentration range, the inhibitory rate was increased with concentration of arachidonic acid and treatment time (P ＜0.05).The positive expression rate of Ki-67 antigen were not significantly changed after treated with 5 and 10 μmol/L of arachidonic acid (P ＞0.05) , and was significantly reduced after treated with 20, 40 and 80 μmol/L of arachidonic acid [(47.1 ±1.6)％, (32.1±4.4)％, (18.6±2.3)％ vs (58.7±4.6)％] (P＜0.05).Conclusion 20-80 μmol/L of arachidonic acid can inhibit the proliferation of human hypertrophic scar fibroblasts, which may be one of the mechanisms of inhibiting hypertrophic scar.