CAJ | 학술논문

目的探讨不同剂量UVA1对全层皮肤缺损诱导的兔耳增生性瘢痕模型的影响的可能机制.方法 24只新西兰白兔双耳腹面手术切除2 cm×5 cm全层皮肤至筋膜建立兔耳增生性瘢痕模型后,随机分成4组,每组6只,将每只兔左耳分别于伤后即刻(U0)、1个月(U1)、2个月(U2)、3个月(U3)开始强功率UVA1照射,右耳为非照射组;各照射组分为两个剂量组[60 J/cm2(M),110 J/cm2(H)],分别在照射前后进行MMP-1、TIMP-1、TGF-β1、PCNA和α-SMA的免疫组织化学染色和透射电镜检查.结果与照射前比较,中高剂量组照射后瘢痕处MMP-1表达:U1组分别为10.43±1.61、11.16±1.57;U2组分别为8.63±2.61、7.33±1.58;U3组分别为5.74±1.43、3.11±0.27;均显著增加(P＜0.05).TGF-β1表达:U1组分别为12.51±4.13、12.02±5.02;U2组分别为18.74±6.42、19.69±4.52;U3组分别为20.51±1.78、29.45±6.55.PCNA表达:U1组分别为2.67±0.44、2.04±0.65;U2组分别为4.50±0.97、5.82±0.68;U3组分别为7.45±1.47、8.16±1.07;均显著降低(P＜0.05).只有高剂量组显著降低TIMP-1表达,U1组为12.74±4.58,U2组为15.17±3.26,U3组为20.72±3.31(P＜0.05).只有U1H、U1M、U2H组α-SMA表达(1.33±0.34、2.04±0.20、3.60±1.75)显著降低(P＜0.05).U0组:与对照组比较,高剂量组MMP-1表达(2.25±0.38)显著降低(P＜0.05),而TGF-β1表达(23.90±2.92)显著增加(P＜0.05).中、高剂量组PCNA(7.42±0.65、7.59±0.31)、TIMP-1(29.82±1.94、33.51±1.19)及α-SMA表达(6.31±0.61、2.97±0.56)均显著增加(P＜0.05).透射电镜结果显示:强功率UVA1照射后胶原纤维直径变细;成纤维细胞胞质变少,细胞器欠发达,多数为静止的纤维细胞.结论 UVA1对瘢痕的作用可能与其抑制TGF-β1、TIMP-1、PCNA和α-SMA的表达,同时促进MMP-1的表达,从而促进基质蛋白降解及抑制成纤维细胞和肌成纤维细胞的增值活性有关.如果UVA1过早干预,则会出现相反的结果.
목적 탐토불동제량UVA1대전층피부결손유도적토이증생성반흔모형적영향적가능궤제.방법 24지신서란백토쌍이복면수술절제2 cm×5 cm전층피부지근막건립토이증생성반흔모형후,수궤분성4조,매조6지,장매지토좌이분별우상후즉각(U0)、1개월(U1)、2개월(U2)、3개월(U3)개시강공솔UVA1조사,우이위비조사조;각조사조분위량개제량조[60 J/cm2(M),110 J/cm2(H)],분별재조사전후진행MMP-1、TIMP-1、TGF-β1、PCNA화α-SMA적면역조직화학염색화투사전경검사.결과 여조사전비교,중고제량조조사후반흔처MMP-1표체:U1조분별위10.43±1.61、11.16±1.57;U2조분별위8.63±2.61、7.33±1.58;U3조분별위5.74±1.43、3.11±0.27;균현저증가(P＜0.05).TGF-β1표체:U1조분별위12.51±4.13、12.02±5.02;U2조분별위18.74±6.42、19.69±4.52;U3조분별위20.51±1.78、29.45±6.55.PCNA표체:U1조분별위2.67±0.44、2.04±0.65;U2조분별위4.50±0.97、5.82±0.68;U3조분별위7.45±1.47、8.16±1.07;균현저강저(P＜0.05).지유고제량조현저강저TIMP-1표체,U1조위12.74±4.58,U2조위15.17±3.26,U3조위20.72±3.31(P＜0.05).지유U1H、U1M、U2H조α-SMA표체(1.33±0.34、2.04±0.20、3.60±1.75)현저강저(P＜0.05).U0조:여대조조비교,고제량조MMP-1표체(2.25±0.38)현저강저(P＜0.05),이TGF-β1표체(23.90±2.92)현저증가(P＜0.05).중、고제량조PCNA(7.42±0.65、7.59±0.31)、TIMP-1(29.82±1.94、33.51±1.19)급α-SMA표체(6.31±0.61、2.97±0.56)균현저증가(P＜0.05).투사전경결과현시:강공솔UVA1조사후효원섬유직경변세;성섬유세포포질변소,세포기흠발체,다수위정지적섬유세포.결론 UVA1대반흔적작용가능여기억제TGF-β1、TIMP-1、PCNA화α-SMA적표체,동시촉진MMP-1적표체,종이촉진기질단백강해급억제성섬유세포화기성섬유세포적증치활성유관.여과UVA1과조간예,칙회출현상반적결과.
Objective To study the possible mechanisms of influence of different doses of UVA1 on the development of hypertrophic scar in rabbit ears induced by excision of full-thickness skin. Methods A hypertrophic scar model was established by excision of full-thickness skin on the ventral surface of rabbit ears.A total of 24 New Zealand white rabbits were randomly and equally divided into 4 groups to receive UVA1 radiation on the left ear immediately (U0 group), 1 month (U1 group), 2 months (U2 group) and 3 months (U3 group) after the excision, respectively, and each group were classified into two subgroups to be irradiated with UVA1 of 60 (middle) and 110 (high) J/cm2, respectively, for 30 sessions. The right ears served as the control without irradiation. Skin samples were obtained from the ears of rabbits before the first and after the last irradiation, transmission electron microscopy (TEM) was used to observe the ultra-structure and morphology of collagen fiber and fibroblasts, and immunohistochemical staining was performed to measure the expressions of matrix metalloproteinases (MMP)-1, tissue inhibitor of metalloproteinase (TIMP)-1, transforming growth factor (TGF)-β1, proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) in skin samples. Results Compared with the unirradiated skin, irradiated skin showed higher expression levels of MMP-1 (P ＜ 0.05), which were 10.43 ± 1.61 and 11.16 ± 1.57 in middle- and high-U1 group, 8.63 ± 2.61 and 7.33 ± 1.58 in middle- and high-U2 gorup, 5.74 ± 1.43 and 3.11 ± 0.27 in middle- and high-U3 group respectively. The expression level of TGF-β1 in irradiated skin was 12.51 ± 4.13 and 12.02 ± 5.02 in middle- and high-U1 group, respectively, 18.74 ± 6.42 and 19.69 ± 4.52 in middle- and high-U2 group, respectively, 20.51 ± 1.78 and 29.45 ± 6.55 in middle- and high-U3 group, respectively. A significant decrease was observed in the expression of PCNA in irradiated skin in middle- and high-U1 group (2.67 ± 0.44 and 2.04 ± 0.65), middle- and high-U2 group (4.50 ± 0.97 and 5.82 ± 0.68), middle- and high-U3 group (7.45 ± 1.47 and 8.16 ±1.07) in comparison with unirradiated skin (all P＜ 0.05). There was a lower expression of TIMP-1 in irradiated skin of high-U1, -U2, and -U3 group (12.74 ± 4.58, 15.17 ± 3.26, 20.72 ± 3.31, all P＜ 0.05) as well as α-SMA in that of high-U1, middle-U1 and high-U2 group (1.33 ± 0.34, 2.04 ± 0.20, 3.60 ± 1.75, all P＜ 0.05) compared with the unirradiated skin. Further more, a significant increment was observed in the expressions of TGF-β1 (23.90 ± 2.92, P ＜ 0.05) in irradiated skin of high-U0 group, PCNA(7.42 ± 0.65 and 7.59 ± 0.31 ),TIMP-1 (29.82 t 1.94 and 33.51 ± 1.19) and α-SMA (6.31 ± 0.61 and 2.97 ± 0.56) in irradiated skin of middle- and high-U0 group, but a decline in the expression of MMP-1 (.25 ± 0.38, P＜ 0.05) in irradiated skin of high-U0 group in comparison with the unirradiated skin. TEM showed that the collagen fiber diameter turned small, and fibroblasts, most of which were quiescent, showed a reduction in cytoplasm volume with the presence of immature organelles, after high-dose UVA1 irradiation. Conclusions The therapeutical effect of UVA1 on scar may be realized by accelerating the degradation of matrix proteins and decelerating the proliferation of fibroblasts and myofibroblasts via downregulating the expressions of TGF-β1, TIMP-1 and α-SMA and upregulating the expression of MMP-1. However, the results would be opposite if the interference with UVA1 irradiation is given at the early stage of wound healing.