论文部分内容阅读
目的研究低氧对系统性硬化病(SSc)患者和正常人皮肤成纤维细胞胶原蛋白合成及胶原基因表达的影响。方法分别以5例SSc患者和5例正常人皮肤成纤维细胞系作为实验对象。将成纤维细胞分别放入20%正常氧和2%低氧分压环境中培养,用分光光度仪测定细胞培养基上清液中羟脯氨酸含量,并折算成胶原蛋白含量。用Trizol提取细胞总RNA,用RT-PCR方法测定其中Ⅰ型和Ⅲ型前胶原mRNA的表达。结果培养至第6天,低氧分压条件下,SSc患者皮肤成纤维细胞培养基中胶原蛋白含量比正常氧分压条件下显著增加(t=3.97,P=0.0041);正常人皮肤成纤维细胞培养基中胶原蛋白含量此时也显著增加(t=2.63,P=0.0302)。培养至第3天,低氧分压条件下,SSc成纤维细胞Ⅰ型和Ⅲ型前胶原mRNA表达量比正常氧分压条件下均有显著增加(Ⅰ型:t=5.81,P=0.0004;Ⅲ型:t=6.44,P=0.0002);正常人皮肤成纤维细胞此时的Ⅰ型和Ⅲ型前胶原mRNA表达量与SSc患者相似,低氧分压条件下比正常氧分压条件下均有显著增加(Ⅰ型:t=4.40,P=0.0023;Ⅲ型:t=2.24,P=0.0453)。结论低氧状态下,SSc患者和正常人皮肤成纤维细胞胶原蛋白合成均有增加,Ⅰ型和Ⅲ型前胶原mRNA表达也均增加.低氧可能是皮肤硬化的一个相关因素。
Objective To investigate the effects of hypoxia on collagen synthesis and collagen gene expression in human dermal fibroblasts from patients with systemic sclerosis (SSc). Methods 5 cases of SSc patients and 5 normal human dermal fibroblasts were used as experimental subjects. Fibroblasts were cultured in the environment of 20% normal oxygen and 2% hypoxic partial pressure respectively. The content of hydroxyproline in the cell culture supernatant was measured by spectrophotometer and converted into collagen content. Total RNA was extracted with Trizol and the expression of procollagen type ¢ ñ and ¢ ó procollagen mRNA was determined by RT-PCR. Results On the 6th day, the collagen content in the skin fibroblasts of SSc patients increased significantly under the condition of hypoxic partial pressure (t = 3.97, P = 0.0041) compared with the normal oxygen partial pressure. Normal human skin fibroblasts Collagen content in the cell culture medium also increased significantly at this time (t = 2.63, P = 0.0302). On day 3, the mRNA expression of type Ⅰ and type Ⅲ procollagen of SSc fibroblasts increased significantly under hypoxic partial pressure (type Ⅰ: t = 5.81, P = 0.0004; Ⅲ type: t = 6.44, P = 0.0002). The mRNA expression of type Ⅰ and type Ⅲ procollagen in normal human dermal fibroblasts was similar to those in patients with SSc at hypoxic partial pressure There was a significant increase (type I: t = 4.40, P = 0.0023; type III: t = 2.24, P = 0.0453). Conclusion Under hypoxia conditions, the collagen synthesis of the skin fibroblasts in both normal and SSc patients increased, and the expressions of type Ⅰ and type Ⅲ procollagen mRNAs also increased.Hypoxemia may be a related factor of skin sclerosis.