转化生长因子β_1对肾间质成纤维细胞胶原表达的影响

An experimental study of the effect of TGF β_1 on the collagen expression by renal interstitial fibroblasts

目的 探讨肾间质纤维化的机制。方法 体外培养大鼠肾间质成纤维细胞，采用Ｌ３Ｈ脯氨酸掺入法测定胶原合成及ＲＴＰＣＲ 技术观察转化生长因子β（ＴＧＦβ１） 对体外培养的大鼠肾间质成纤维细胞Ⅰ和Ⅲ型胶原ｍＲＮＡ 表达的影响。结果 ＴＧＦβ１ 可促进肾间质成纤维细胞的３Ｈ脯氨酸掺入，且随着ＴＧＦβ１ 剂量的增加，３Ｈ脯氨酸掺入量增加，胶原合成增多。ＴＧＦβ１ 可促进Ⅰ和Ⅲ型胶原ｍＲＮＡ 的表达，且在ＴＧＦβ１ 浓度２ ～１０ ｎｇ／ｍｌ 时ｍＲＮＡ 的表达量与浓度呈现正相关；在６ ～４８小时内与时间呈现正相关。结论 ＴＧＦβ１ 可能通过促进肾间质成纤维细胞胶原的合成及ｍＲＮＡ 的表达而参与肾间质纤维化。

Objective To investigate the effect of TGF β 1 on the proliferation, collagen synthesis and collagen mRNA type Ⅰand type Ⅲ expression of renal interstitial fibroblasts in vitro. Methods Fibroblasts were isolated from rat renal medulla. Collagen synthesis and expression of collagen type Ⅰand type Ⅲ mRNA of renal interstitial fibroblasts were investigated by L 3 H proline incorporation and RT PCR, respectively. Results TGF β 1 had a dose dependent stimulating effect on the collagen synthesis of fibroblasts with or without the presence of serum (r= 0 9822, P<0 05 and r = 0 9597, P<0 05, respectively). TGF β 1 could stimulate the proliferation and collagen synthesis of renal interstitial fibroblasts. By the RT PCR method,TGF β 1 could increase the expression of collagen type Ⅰ and type Ⅲ mRNA of fibroblasts with dose dependent (from 2 to 10 ng/ml) and time dependent effects (from 6 to 48 hours with the concentration of TGF β 1 of 5 ng/ml).Conclusion TGF β 1 at a low concentration may be involved in renal interstitial fibrosis by promoting the collagen synthesis and expression of fibroblasts.