摘要
背景:研究表明,转化生长因子β2在青光眼滤过性手术后滤过泡瘢痕形成过程中起着重要的作用。目的:实验拟观察抗转化生长因子β2抗体对体外培养的滤过泡成纤维细胞增殖及p21、cyclinD1表达的影响。设计、时间及地点:开放性实验,于2007-06在广东医学院附属医院整形科完成。材料:取自青光眼小梁切除术后失败滤过泡进行手术修复时切除下来的瘢痕组织。患者均为男性,年龄23～35岁。8例患者术前、术中均未接受抗瘢痕药物治疗。方法:将瘢痕组织进行原代细胞培养,获得成纤维细胞。将浓度为0(对照),0.01,0.1,1,10mg/L抗转化生长因子β2抗体作用于成纤维细胞24h,半数抑制浓度(IC50)抗转化生长因子β2抗体分别处理0(对照),24,48,72h。主要观察指标:四甲基偶氮唑盐法及免疫组织化学法观察抗转化生长因子β2抗体对成纤维细胞的增殖及p21、cyclinD1表达的作用。结果:①随着抗转化生长因子β2抗体作用浓度的增加及作用时间的延长,抗体对成纤维细胞具有明显抑制作用。②随着抗转化生长因子β2抗体作用浓度、时间增加cyclinD1阳性表达减少,弱阳性表达增加;而p21强阳性表达明显增多,且在细胞核的表达也增多(P<0.01);在不同的作用浓度及时间,p21与cyclinD1表达呈显著负相关(r=-0.91,r=-0.85,P<0.05),两者与细胞的增殖抑制显著相关(r=0.76,P<0.05)。结论:抗转化生长因子β2抗体可能通过诱导p21蛋白合成,下调cyclinD1的表达从而抑制成纤维细胞的增殖。
BACKGROUND： Researches indicate that transforming growth factor-β2 plays an important role in the cicatrisation of filtering bleb after glaucoma filtering surgery.
OBJECTIVE： To observe the influence of anti-transforming growth factor-β2 （TGF-β2） on the proliferation of filtering bleb fibroblasts cultured in vitro and its expression of p21 and cyclinD1.
DESIGN, TIME AND SETTING： The open experiment was carried out in June 2007 at Department of Plastic Surgery, the Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China.
MATERIALS： Scar tissues removed from failed filtering blebs after glaucoma trabeculectomy. All patients were male and their age ranged from 23 to 35 years; eight patients did not receive anti-scar drug treatment before and during the operation.
METHODS： Primary cell culture was done using the scar tissue to get fibroblasts. Fibroblasts were incubated with 0 （control）, 0.01, 0.1, 1, 10 mg/L of anti-TGF-β2 antibody for 24 hours; inhibitory concentration 50 of anti-TGF-β2 antibody was used to treat the fibroblasts for 0 （control）, 24, 48, 72 hours, respectively.
MAIN OUTCOME MEASURES： The influence of anti-TGF-β2 antibody on the proliferation of fibroblasts and its expression of p21 and cyclinD1 observed with MTT and immunohistochemistry.
RESULTS： With the increase of the anti-TGF-β2 antibody concentration and the prolongation of the action time, the inhibitory effect of antibody on the fibroblasts became more and more obvious; the cyclinD1 positive expression decreased gradually while the weak positive expression increased; on the other hand, the strong positive expression of p21 raised noticeably and the expression in cell nucleus also increased （P 〈 0.01）. The expression of p21 showed a marked negative correlation with the expression of cylinD1 under different concentrations of anti-TGF-β2 antibody and at different time points （r=-0.91, r=-0.85, P 〈 0.05）, p21 and cylinD1 both were correlated noticeably with the proliferation inhibition of cells （r=-0.76, P 〈 0.05）.
CONCLUSION： The anti-TGF-β2 antibody can inhibit the proliferation of fibroblasts by increasing the synthesis of p21 protein and regulating the expression of cyclinD1.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第28期5401-5405,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
湛江市科技攻关项目(2006C01019)~~
