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背景:肌腱组织细胞具有分泌胶原的功能,在伤口愈合及粘连中有重要的作用。知之较少的是乳酸对肌腱细胞的生物学作用。目的:探讨兔屈趾肌腱腱鞘、腱外膜和腱内膜细胞增殖、胶原产生和乳酸对细胞的增殖、胶原产生和对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8分泌的影响。设计、时间和地点:随机对照动物实验,于2005-09/2006-07在青岛大学医学院附属医院动物实验中心完成。材料:选用6只成年清洁级新西兰大白兔,雌雄不拘。方法:①进行兔腱鞘成纤维细胞、腱外膜和腱内膜成纤维样细胞的分离和培养,亚甲蓝染色观察细胞形态并鉴定3种细胞类型。②以加入25mmol/L乳酸培养细胞,测量细胞的数量。③比较加入25,50,100和200mmol/L乳酸后细胞胶原产生量和对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8分泌量,并与不加乳酸培养为对照比较。主要观察指标:使用免疫组化检测Ⅰ、Ⅱ、Ⅲ型胶原的产生,通过酶联免疫吸附试验定量检测不同剂量乳酸对细胞Ⅰ型胶原产生的影响及对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8产生的影响。结果:①25mmol/L乳酸使培养的细胞数量降低,3种细胞组间比较差异无显著性意义(P>0.05)。②乳酸可以使实验组Ⅰ、Ⅱ和Ⅲ型胶原组织产生明显多于对照组,差异有显著性意义(P<0.05)。③当乳酸浓度增加至50mmol/L时,实验组3种细胞的Ⅰ型胶原量增加达最高峰,与乳酸浓度为0mmol/L时比较,差异有非常显著性意义(t=4.58,3.97,3.16,P<0.01);当乳酸浓度增加到100mmol/L和200mmol/L时,Ⅰ型胶原产生量明显减少。④实验组乳酸浓度为25mmol/L时对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8的分泌量增加,白细胞介素8的分泌量减少,与对照组比较差异均有显著性意义(P<0.05)。结论:乳酸能增加腱鞘成纤维细胞、腱外膜细胞和腱内膜细胞的胶原产生量,增加的程度与乳酸的浓度有关,以50mmol/L时最佳,而这种刺激作用可能与增加转化生长因子β1,基础纤维母细胞生长因子和减少白细胞介素8的分泌量有关。
Background: Tendon cells have the function of secreting collagen and play an important role in wound healing and adhesion. Little is known about the biological effects of lactate on tendon cells. OBJECTIVE: To investigate the proliferation, collagen production and cell proliferation, collagen production and the effects of lactic acid on transforming growth factor β1, basal fibroblast growth factor, interleukin-8 secreted by rabbit tendon sheath, tendon adventitia and tendon endometrium influences. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Animal Experimental Center, Affiliated Hospital of Qingdao University Medical College from September 2005 to July 2006. Material: 6 adult clean grade New Zealand white rabbits, both male and female. Methods: ①The tendon sheath fibroblasts, tendon adventitia and tendon endomembrane fibroblast-like cells were isolated and cultured. The cell morphology was observed by methylene blue staining and three cell types were identified. ② Add 25mmol / L lactic acid cultured cells, the number of cells measured. ③ Comparison of the amount of collagen produced by 25, 50, 100 and 200 mmol / L lactic acid and the secretion of transforming growth factor β1, basic fibroblast growth factor and interleukin 8 were compared with those without lactic acid. MAIN OUTCOME MEASURES: The generation of collagen type Ⅰ, Ⅱ and Ⅲ was detected by immunohistochemistry. The effects of different doses of lactic acid on the type Ⅰ collagen production and the effects on the growth of transforming growth factor β1 and basic fibroblasts were quantitatively detected by enzyme-linked immunosorbent assay Factors, the impact of interleukin-8 production. Results: ①25mmol / L lactic acid decreased the number of cultured cells, the difference between the three cell groups was not significant (P> 0.05). ② Lactic acid could significantly increase the expression of collagen type Ⅰ, Ⅱ and Ⅲ in the experimental group compared with the control group (P <0.05). ③ When the concentration of lactic acid increased to 50mmol / L, the amount of type I collagen in the three experimental groups increased at the peak, which was significantly different from that of the concentration of lactic acid at 0mmol / L (t = 4.58,3.97,3.16 , P <0.01). When the concentration of lactic acid was increased to 100 mmol / L and 200 mmol / L, the production of type I collagen was significantly reduced. ④ When the concentration of lactic acid in experimental group was 25mmol / L, the secretion of transforming growth factor-β1, basic fibroblast growth factor and interleukin-8 increased and the secretion of interleukin-8 decreased, which was significantly different from the control group Significance (P <0.05). CONCLUSION: Lactic acid can increase collagen production in tendon sheath fibroblasts, tendon adventitial cells and tendon endothelium cells, and the degree of increase is related to the concentration of lactic acid. The optimal concentration of lactic acid is 50 mmol / L, which may be associated with increased conversion Growth factor β1, basal fibroblast growth factor and reduce the secretion of interleukin 8 related.