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.
揹景:哺乳類動物有3種TGF-β異構體即TGF-β1,β2,β3,這3種異構體各自具有獨特而不同的生物學作用,TGF-β1是明顯的促瘢痕形成因子,而TGF-β3可減少TGF-β1,β2的產生,從而減少細胞外基質(ECM)閤成,因此,TGF-β3有可能是人體內天然的抗瘢痕形成因子.目的:通過外源性TGF-β3對增生性瘢痕成纖維細胞(HSFB)生長增殖及Ⅰ,Ⅲ型膠原及TGF-β1蛋白錶達的影響,瞭解其生物學行為,為臨床治療提供理論依據.設計:隨機對照實驗研究.地點和對象:汕頭大學醫學院第二附屬醫院整形燒傷外科完成,對象為新鮮無菌增生性瘢痕組織,來源于本科收治及門診手術的增生性瘢痕患者6例,男3例,女3例;年齡5～45歲.榦預:6例增生性瘢痕成纖維細胞體外培養,分為4組,實驗組又分為5,10,50 mg/L組分彆加TGF-β3 5,10,及50 mg/L,對照組加入FBM1.5 mL,採用MTT比色法測定TGF-β3對HSFB增殖活性的影響,3H-脯氨痠摻人法測定其膠原閤成,免疫組化檢測Ⅰ,Ⅲ型膠原及TGF-β1蛋白錶達情況.主要觀察指標:TGF-β3對HSFB體外增殖,HSFB Ⅰ,Ⅲ型膠原及TGF-β1蛋白錶達的影響,TGF-β3作用後HSFB膠原閤成情況.結果:轉化生長因子β3可抑製HSFB細胞增殖,作用120 h後實驗組細胞密度分彆為(6.34±0 51),(6.01±0.38),(5.24±0.65)×105/瓶,明顯低于對照組(10.69±0.76)x105/瓶(F=102.432～163 024,P＜0.01);轉化生長因子β3可抑製膠原蛋白閤成,實驗組(10,50mg/L組)脯氨痠含量分彆為(164.01±70.27),(151 02±49 85)min-1明顯低于對照組9231.56±67.83)min-1(q=32.124,31.021,P＜0.01);下調TGF-β1蛋白及Ⅰ型膠原蛋白錶達;而對Ⅲ型膠原影響較小.結論:TGF-β3可抑製HSFB細胞增殖,下調TGF-β1蛋白錶達,減少膠原閤成,顯示其具有抗組織纖維化的作用,具有臨床應用價值.
배경:포유류동물유3충TGF-β이구체즉TGF-β1,β2,β3,저3충이구체각자구유독특이불동적생물학작용,TGF-β1시명현적촉반흔형성인자,이TGF-β3가감소TGF-β1,β2적산생,종이감소세포외기질(ECM)합성,인차,TGF-β3유가능시인체내천연적항반흔형성인자.목적:통과외원성TGF-β3대증생성반흔성섬유세포(HSFB)생장증식급Ⅰ,Ⅲ형효원급TGF-β1단백표체적영향,료해기생물학행위,위림상치료제공이론의거.설계:수궤대조실험연구.지점화대상:산두대학의학원제이부속의원정형소상외과완성,대상위신선무균증생성반흔조직,래원우본과수치급문진수술적증생성반흔환자6례,남3례,녀3례;년령5～45세.간예:6례증생성반흔성섬유세포체외배양,분위4조,실험조우분위5,10,50 mg/L조분별가TGF-β3 5,10,급50 mg/L,대조조가입FBM1.5 mL,채용MTT비색법측정TGF-β3대HSFB증식활성적영향,3H-포안산참인법측정기효원합성,면역조화검측Ⅰ,Ⅲ형효원급TGF-β1단백표체정황.주요관찰지표:TGF-β3대HSFB체외증식,HSFB Ⅰ,Ⅲ형효원급TGF-β1단백표체적영향,TGF-β3작용후HSFB효원합성정황.결과:전화생장인자β3가억제HSFB세포증식,작용120 h후실험조세포밀도분별위(6.34±0 51),(6.01±0.38),(5.24±0.65)×105/병,명현저우대조조(10.69±0.76)x105/병(F=102.432～163 024,P＜0.01);전화생장인자β3가억제효원단백합성,실험조(10,50mg/L조)포안산함량분별위(164.01±70.27),(151 02±49 85)min-1명현저우대조조9231.56±67.83)min-1(q=32.124,31.021,P＜0.01);하조TGF-β1단백급Ⅰ형효원단백표체;이대Ⅲ형효원영향교소.결론:TGF-β3가억제HSFB세포증식,하조TGF-β1단백표체,감소효원합성,현시기구유항조직섬유화적작용,구유림상응용개치.
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.
