BACKGROUND: In mammals, there are three types of isomers of transforming growth factor-β(TGF-β), i. e. TGF-β1, β2 and β3. All of them have specific and different biological effects. TGF-β1 is a well recognized epulotic factor, while TGF-β3 can reduce the production of TGF-β1 and β2 and thus decease the synthesis of extracellular matrix(ECM). Therefore, TGF-β3 may be the natural factor that resists scar formation in human body.OBJECTIVE: To investigate the biological behavior of TGF-β3 by studying the influence of exogenous TGF-β3 on the growth and proliferation of hyperplastic scar fibroblast(HSFB) as well as its effects on type I and type Ⅲcollagen and the expression of TGF-β1 protein, so as to provide a theoretical basis for clinical treatment,DESIGN: A randomized controlled experimental study was conducted.SETTING and PARTICIPANTS: The study was done in the Department of Burns and Plastic Surgery in the Second Affiliated Hospital of Shantou University Medical College. The samples used were fresh aseptic hyperplastic scar tissues obtained from 6 patients treated at the department and outpatient service center, including 3 males and 3 females (aged 5 -45 years old).INTERVENTIONS: In vitro culture of HSFB was performed in the 6 patients who were divided into 4 groups: The experimental group was divided into 3 groups, in which 5 mg/L, 10 mg/L and 50 mg/L TGF-β3 were used respectively; 1.5 mL FBM was used in control group. Influence of TGF-β3on the proliferation activity of HSFB was measured by MTT colorimetric method aud the synthesis of collagen was measured by 3H-proline incorporative method. Type Ⅰ and type Ⅲ collagen and the expression of TGF-β1protein were detected by immunohistochemical method.MAIN OUTCOME MEASURES: Influence of TGF-β3 on the in vitro proliferation of HSFB, type Ⅰ and type Ⅲ collagen of HSFB and the expression of TGF-β1 protein; collagen synthesis of HSFB resulted from TGF-β3 effect.RESULTS: TGF-β3 could inhibit the proliferation of HSFB. The cell density of the experimental groups was (6.34±0.51), (6.01±0.38),(5.24 ±0. 65) x 105/flask respectively after 120 hours, which was significantly lower than that of control group whose cell density was (10. 69 ±0. 76) × 105/flask ( F = 102. 432 - 163. 024, P ＜ 0.01). TGF-β3 could inhibit the synthesis of collagen. The content of proline was (164.01 ± 70.27)and (151.02 ±49.85)/minute in two experimental groups(10 mg/L,50 mg/L), which was significantly lower than that of control group whose content was(9231.56±67.83)/minute(q=32.124, 31.021, P ＜0.01 ) . TGF-β3 could decrease the expression of TGF-β protein and type Ⅰcollagen and had minor irnfluence on type Ⅲ collagen.%背景:哺乳类动物有3种TGF-β异构体即TGF-β1,β2,β3,这3种异构体各自具有独特而不同的生物学作用,TGF-β1是明显的促瘢痕形成因子,而TGF-β3可减少TGF-β1,β2的产生,从而减少细胞外基质(ECM)合成,因此,TGF-β3有可能是人体内天然的抗瘢痕形成因子.目的:通过外源性TGF-β3对增生性瘢痕成纤维细胞(HSFB)生长增殖及Ⅰ,Ⅲ型胶原及TGF-β1蛋白表达的影响,了解其生物学行为,为临床治疗提供理论依据.设计:随机对照实验研究.地点和对象:汕头大学医学院第二附属医院整形烧伤外科完成,对象为新鲜无菌增生性瘢痕组织,来源于本科收治及门诊手术的增生性瘢痕患者6例,男3例,女3例;年龄5～45岁.干预:6例增生性瘢痕成纤维细胞体外培养,分为4组,实验组又分为5,10,50 mg/L组分别加TGF-β3 5,10,及50 mg/L,对照组加入FBM1.5 mL,采用MTT比色法测定TGF-β3对HSFB增殖活性的影响,3H-脯氨酸掺人法测定其胶原合成,免疫组化检测Ⅰ,Ⅲ型胶原及TGF-β1蛋白表达情况.主要观察指标:TGF-β3对HSFB体外增殖,HSFB Ⅰ,Ⅲ型胶原及TGF-β1蛋白表达的影响,TGF-β3作用后HSFB胶原合成情况.结果:转化生长因子β3可抑制HSFB细胞增殖,作用120 h后实验组细胞密度分别为(6.34±0 51),(6.01±0.38),(5.24±0.65)×105/瓶,明显低于对照组(10.69±0.76)x105/瓶(F=102.432～163 024,P＜0.01);转化生长因子β3可抑制胶原蛋白合成,实验组(10,50mg/L组)脯氨酸含量分别为(164.01±70.27),(151 02±49 85)min-1明显低于对照组9231.56±67.83)min-1(q=32.124,31.021,P＜0.01);下调TGF-β1蛋白及Ⅰ型胶原蛋白表达;而对Ⅲ型胶原影响较小.结论:TGF-β3可抑制HSFB细胞增殖,下调TGF-β1蛋白表达,减少胶原合成,显示其具有抗组织纤维化的作用,具有临床应用价值.
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