中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2012年
4期
357-360
,共4页
白晓智%胡大海%王耘川%刘佳琦%石继红%汤朝武
白曉智%鬍大海%王耘川%劉佳琦%石繼紅%湯朝武
백효지%호대해%왕운천%류가기%석계홍%탕조무
白芷%成纤维细胞%伤口愈合%胶原Ⅰ型%胶原Ⅲ型
白芷%成纖維細胞%傷口愈閤%膠原Ⅰ型%膠原Ⅲ型
백지%성섬유세포%상구유합%효원Ⅰ형%효원Ⅲ형
Angelica dahurica%Fibroblasts%Wound healing%Collagen type Ⅰ%Collagen type Ⅲ
目的 体外观察白芷活性提取物对人皮肤成纤维细胞(Fb)生物学特性的影响,探讨其促进创面愈合的可能机制.方法 分离培养人正常Fb,按照随机数字表法分白芷活性提取物不同浓度组,以体积分数0.25％的新生牛血清DMEM为对照组.采用噻唑蓝比色法(MTT)检测不同浓度白芷活性提取物对体外培养的Fb增殖作用的影响.通过流式细胞仪、real-time PCR分别检测白芷活性提取物最适浓度培养条件下Fb的细胞周期变化以及Ⅰ、Ⅲ型胶原mRNA的表达情况.结果 白芷活性提取物在5×10 -4～5×10- 2g/L浓度范围内可显著促进Fb增殖,呈剂量依赖性,不同浓度白芷活性提物培养液组测得吸光度(A)值分别为0 346±0.055、0.387±0.043和0.310±0.026,与对照组(0.273 ±0.042)比较,差异有统计学意义(t=3.36、5.79和2.49,P＜0.05),其中在5×10-3 g/L浓度时测得A值最高,促增殖作用最为显著,为白芷活性提取物促进Fb增殖的最适合浓度.此浓度条件下白芷活性提取物可明显促进Fb通过G1/S及S/G2期限制点,S期(26.6±2.6)及G2/M期(7.4±1.3)细胞与对照组相比明显增多,G0/G1期细胞(66.3±4.5)与对照组相比明显减少(t=11.2、5.69、2.44,P ＜0.05).5×10 -3g/L白芷活性提取物组与对照组比较Ⅰ、Ⅲ型胶原mRNA表达明显上调(分别为1.61 ±0.26比1.00±0.16和3.36 ±0.40比1.00 ±0.14),差异有统计学意义(t=6.69、7.64,P＜0.01).结论 白芷活性提取物能显著促进Fb增殖,加快Fb细胞周期进程,同时可上调Ⅰ、Ⅲ型胶原mRNA的表达.可能与白芷促进创面愈合的机制有关.
目的 體外觀察白芷活性提取物對人皮膚成纖維細胞(Fb)生物學特性的影響,探討其促進創麵愈閤的可能機製.方法 分離培養人正常Fb,按照隨機數字錶法分白芷活性提取物不同濃度組,以體積分數0.25％的新生牛血清DMEM為對照組.採用噻唑藍比色法(MTT)檢測不同濃度白芷活性提取物對體外培養的Fb增殖作用的影響.通過流式細胞儀、real-time PCR分彆檢測白芷活性提取物最適濃度培養條件下Fb的細胞週期變化以及Ⅰ、Ⅲ型膠原mRNA的錶達情況.結果 白芷活性提取物在5×10 -4～5×10- 2g/L濃度範圍內可顯著促進Fb增殖,呈劑量依賴性,不同濃度白芷活性提物培養液組測得吸光度(A)值分彆為0 346±0.055、0.387±0.043和0.310±0.026,與對照組(0.273 ±0.042)比較,差異有統計學意義(t=3.36、5.79和2.49,P＜0.05),其中在5×10-3 g/L濃度時測得A值最高,促增殖作用最為顯著,為白芷活性提取物促進Fb增殖的最適閤濃度.此濃度條件下白芷活性提取物可明顯促進Fb通過G1/S及S/G2期限製點,S期(26.6±2.6)及G2/M期(7.4±1.3)細胞與對照組相比明顯增多,G0/G1期細胞(66.3±4.5)與對照組相比明顯減少(t=11.2、5.69、2.44,P ＜0.05).5×10 -3g/L白芷活性提取物組與對照組比較Ⅰ、Ⅲ型膠原mRNA錶達明顯上調(分彆為1.61 ±0.26比1.00±0.16和3.36 ±0.40比1.00 ±0.14),差異有統計學意義(t=6.69、7.64,P＜0.01).結論 白芷活性提取物能顯著促進Fb增殖,加快Fb細胞週期進程,同時可上調Ⅰ、Ⅲ型膠原mRNA的錶達.可能與白芷促進創麵愈閤的機製有關.
목적 체외관찰백지활성제취물대인피부성섬유세포(Fb)생물학특성적영향,탐토기촉진창면유합적가능궤제.방법 분리배양인정상Fb,안조수궤수자표법분백지활성제취물불동농도조,이체적분수0.25％적신생우혈청DMEM위대조조.채용새서람비색법(MTT)검측불동농도백지활성제취물대체외배양적Fb증식작용적영향.통과류식세포의、real-time PCR분별검측백지활성제취물최괄농도배양조건하Fb적세포주기변화이급Ⅰ、Ⅲ형효원mRNA적표체정황.결과 백지활성제취물재5×10 -4～5×10- 2g/L농도범위내가현저촉진Fb증식,정제량의뢰성,불동농도백지활성제물배양액조측득흡광도(A)치분별위0 346±0.055、0.387±0.043화0.310±0.026,여대조조(0.273 ±0.042)비교,차이유통계학의의(t=3.36、5.79화2.49,P＜0.05),기중재5×10-3 g/L농도시측득A치최고,촉증식작용최위현저,위백지활성제취물촉진Fb증식적최괄합농도.차농도조건하백지활성제취물가명현촉진Fb통과G1/S급S/G2기한제점,S기(26.6±2.6)급G2/M기(7.4±1.3)세포여대조조상비명현증다,G0/G1기세포(66.3±4.5)여대조조상비명현감소(t=11.2、5.69、2.44,P ＜0.05).5×10 -3g/L백지활성제취물조여대조조비교Ⅰ、Ⅲ형효원mRNA표체명현상조(분별위1.61 ±0.26비1.00±0.16화3.36 ±0.40비1.00 ±0.14),차이유통계학의의(t=6.69、7.64,P＜0.01).결론 백지활성제취물능현저촉진Fb증식,가쾌Fb세포주기진정,동시가상조Ⅰ、Ⅲ형효원mRNA적표체.가능여백지촉진창면유합적궤제유관.
Objective To observe the effects of Angelica dahurica extracts on the biological characteristics of human dermal fibroblasts in vitro and to preliminary explore its possible therapeutic mechanism for wound healing.Methods The optimal concentration of Angelica dahurica extracts was identified by analysing of proliferation activity of human normal fibroblasts (Fb) that treated with different concentration of Angelica dahurica extracts through thiazole blue (MTT) colorimetric assay.Cell cycle,collagen Ⅰ and collagen Ⅲ mRNA levels of the optimal Angelica dahurica extracts treated Fb were detected by flow cytometry ( FCM ) and real-time PCR techniques.Results At concentrations of 5 × 10 -4 to 5 ×10- 2 g/L,the Angelica dahurica extracts significantly enhanced the proliteration of Fb.The most significant concentration was 5 × 10-3 g/L (t =5.79,P ＜0.01 ),at which an increased percentage of G1 to S and S to G2 phase cells ( t =11.2,5.69,2.44,P ＜ 0.05 ) as well as an increased level of collagen Ⅰ ( 1.61 ±0.26 vs.1.00 ±0.16) and collagen Ⅲ mRNA (3.36 ±0.40 vs.1.00 ±0.14) were obtained compared to the control group (t =6.69,7.64,P ＜0.01 ).Conclusions Angelica dahurica extracts can notably promote the proliferation of Fb and accelerating the cell cycle of Fb as well as up-regulating thc cxpression of collagen Ⅰ and collagen Ⅲ,which may enhance the process of wound healing.
