乳酸对肌腱细胞转化生长因子-β表达的调节作用

Effects of lactate on TGF-β expression of flexor tendon cells

目的探讨乳酸对兔屈趾肌腱腱鞘、腱外膜和腱内膜细胞转化生长因子(TGF)-β及其受体产生的影响。方法从兔屈趾肌腱分离腱鞘、腱外膜和腱内膜细胞并分别进行培养,在使用25mmol/L的乳酸培养后,酶联免疫吸附试验定量检测TGF-β及其受体的表达,同时应用原位杂交和免疫组织化学技术测量TGF-β1的表达。结果乳酸能显著增加3种细胞所有TGF-β及其受体的表达(P<0.05),其中腱鞘细胞的TGF-β1和TGF-β2增加值最大,腱外膜细胞的TGF-β1和TGF-β2的受体增加值最大,腱内膜细胞则为TGF-β3增加值最大;乳酸还显著增加3种细胞的TGF-β1表达。结论乳酸能显著增加肌腱细胞的TGF-β、TGF-β受体和TGF-β1 mRNA的表达,因此为肌腱愈合过程中调节TGF-β水平提供新的途径。

Objective To study the effects of lactate on TGF-β peptide and receptor production by flexor tendon cells. Methods Tendon sheath fibroblasts, epitennon tenocytes, and endotendanon tenocytes were isolated from rabbit flexor tendon and cultured separately. Cell cultures were supplemented with lactate and the expression of three TGF-β peptide isoforms （ β1, β2 and β3 ） and three receptor isoforms （ R1, R2 and R3 ） was quantified by enzyme-linked immunosorbent assay. The expression of TGF-β1 was also detected by using in situ hybridization and immunohistochemistry. Results Supplementation of the cell culture medium with lactate significantly increased the expression of all TGF-β peptide and receptor isoforms in all three cell lines （ P 〈 0.05 ）. Tendon sheath fibroblasts exhibited the greatest increases in β1 and β2 peptide isoform expression,whereas epitenon tenocytes exhibited the greatest increases in receptor isoform R1 and R2 expression, endotenon tenocytes demonstrated the greatest increase in β3 peptide expression. All three tendon cell types demonstrated significant increases in TGF-β1 mRNA expression when exposed to lactate （ P 〈 0.05 ）. Conclusion Lactate significantly increased the expression of TGF-β peptide,TGF-β receptor and TGF-β mRNA. Modulation of lactate levels may provide a means of modulating the effects of TGF-β on adhesion formation in flexor tendon wound healing.