压应力对增生性瘢痕成纤维细胞增殖与凋亡的影响  

EFFECT OF COMPRESSIVE STRESS ON PROLIFERATION AND APOPTOSIS OF HUMAN HYPERPLASTIC SCAR FIBROBLASTS IN VITRO

作　　者：肖洪[1] 李建福[1] 

机构地区：[1]第三军医大学西南医院整形科,重庆400038

出　　处：《中国修复重建外科杂志》2007年第12期1330-1334,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30570708);国家重点基础研究发展计划(973)资助项目(C12005gb5226037)~~

摘　　要：目的探讨压应力对体外培养人增生性瘢痕成纤维细胞增殖和凋亡的影响。方法应用组织块培养法获取人增生性瘢痕成纤维细胞。以简易气控加压细胞培养仪给细胞施5、10、15、25、50、100、150mmHg(1mmHg=0.133kPa)压力(n=6),持续4d,作为实验组;不施压设为对照组(n=6)。分别以MTT法测定细胞吸光度(A)值并计算生长抑制率(inhibitionratio,IR),流式细胞仪测定细胞生长周期及Annexin-V-PI标记法测定细胞凋亡情况。结果5、10、15、25、50、100、150mmHg压力组及对照组,细胞A值分别为0.228±0.004、0.226±0.003、0.213±0.005、0.180±0.005、0.172±0.007、0.165±0.004、0.164±0.004、0.230±0.005,IR分别为0.8%、2.0%、7.3%、21.7%、25.2%、28.2%、28.2%和0。DNAG1期细胞百分比分别为71.80%±0.44%、72.32%±0.40%、74.56%±1.01%、82.82%±2.76%、86.77%±2.06%、88.23%±1.27%、89.11%±1.74%、71.46%±0.49%,早期细胞凋亡率分别为4.22%±0.49%、5.12%±0.74%、8.58%±0.79%、19.28%±1.40%、25.60%±1.21%、35.80%±2.39%、36.18%±2.38%、4.00%±0.36%。其中5、10mmHg压力组上述各指标与对照组比较,差异无统计学意义(P>0.05);15、25、50、100、150mmHg压力组各指标与对照组比较,差异均有统计学意义(P<0.05);10、15、25、50mmHg压力组细胞A值和G1期细胞百分比组间比较,差异均有统计学意义(P<0.01),50、100、150mmHg压力组各组间比较,差异无统计学意义(P>0.05);10、15、25、50、100mmHg压力组早期细胞调亡率组间比较差异均有统计学意义(P<0.01),100、150mmHg压力组组间比较差异无统计学意义(P>0.05)。结论适当的持续压应力对增生性瘢痕成纤维细胞具有诱导凋亡、抑制增殖的复合效应,是临床上压迫疗法治疗增生性瘢痕的重要机制之一。Objective To investigate an effect of compressive stress on proliferation and apoptosis of human hyperplastic scar fibroblasts（HSFb） in vitro. Methods HSFb were obtained from a 20-year-old female patient who developed a hyperplastic scar 3 months after operation for a large-area burn. HSFb were isolated, and were cultured in vitro with the simplified air-pressure controlled cell-culture instrument, and then they were randomly divided into the following 8 groups., the control group （no stress） and the 7 continuous compressive stress groups, which respectively underwent the 5, 10, 15, 25, 50, 100 and 150 mmHg（1 mmHg=0.133 kPa）pressure treatment for 4 days. The absorbance （A） of the cell and the inhibition ratio （IR） of the cell proliferation were determined by the MTT assay, the cell growth cycle was determined by the flow cytometer, and the cell apoptosis was observed by the Annexin V binding with PI labeling method. Results In the 5, 10, 15, 25, 50, 100 and 150 mmHg pressure groups and the control group, the A values of the cells were 0.228±0.004, 0.226±0.003, 0.213±0.005, 0. 180±0.005, 0. 172±0.007, 0. 165± 0. 004, 0. 164 ± 0. 004 and 0. 230±0. 005, respectively; the IRs of the cell proliferation were 0.8% ,2.0%,7.3%,21.7%,25.2%, 28.2% and 0, respectively; the ratios of the cells in G1 were 71.80% ± 0.44%, 72 0.40%, 74.56%±1.01%, 82.82%±2.76%, 86.77%±2.06%, 88.23%±1.27%, 89.11%±1.74% and 71 0.49%, respectively; the cell apoptosis ratios were 4.22%±0.49%, 5.12%±0.74 %, 8.58%±0.79 %, 19 2.0%, 32%± 46%±28%±1.40%, 25.60%±1.21%, 35.80%± 2.39%, 36.18%±2.38% and 4.00%±0.36%, respectively. In the 5 and 10 mmHg groups there were no statistically significant differences in all the above parameters when compared with those in the control group （P〉0.05）; however, in the 15, 25, 50, 100 and 150 mmHg groups there were statistically significant differences in the above parameters when compared with those in the control group （P〈0.05）.Furthermore. in the 10, 15. 2,5 and 50

关 键 词：机械压应力 增生性瘢痕 成纤维细胞 增殖 凋亡 体外 

分 类 号：R622[医药卫生—整形外科]