Effects of proliferation of rat hepatic stellate cells and collagen-Ⅰ production by antisense oligodeoxynucleotides against platelet-derived growth factor B-chain / 中国病理生理杂志

ABSTRACT

AIM: To investigate the effect of antisense oligodeoxynucleotides against platelet-derived growth factor B-chain(PDGF-B) in the proliferation of cultured rat hepatic stellate cells and collagen-Ⅰ production.METHODS: The phosphorothioate antisense oligodeoxynucleotides and missense oligodeoxynucleotides were synthesized.The rat hepatic stellate cells(HSCs) were treated with these oligodeoxynucleotides at different concentrations,respectively.The inhibition of the proliferating of cultured rat hepatic stellate cells was assayed by MTT method.The expressions of PDGF-B and collagenⅠ mRNA were detected by means of reverse transcription-polymerase chain reaction(RT-PCR).The expression of endogenous PDGF-B was determined by flow cytometry(FCM).Collagen-Ⅰin the supernatant was determined by ELISA.RESULTS: The results showed that the ASODN against the PDGF-B at a final concentration of 10 ?mol/L inhibited the HSCs proliferation,the expressions of PDGF-B and collagen-ⅠmRNA.FCM and ELISA demonstrated that the expression of endogenous PDGF-B and the production of collagen-Ⅰin HSCs were significantly lower than those in controls,whereas the controls(missense oligodeoxynucleotides) had no effect at the same concentration.CONCLUSION:PDGF-B ASODN inhibits the proliferation of cultured rat hepatic stellate cells,the endogenous expression of PDGF-B and the production of collagen.PDGF-B ASON might be useful for gene therapy in liver fibrosis.