Effects of interfering histone demethylase KDM3A expression on hepatic stellate cell proliferation and collagen synthesis
Objective To investigate the impact of interfering with the expression of histone demethy-lase KDM3A on the proliferation and collagen synthesis of hepatic stellate cells(HSCs).Methods Primary rat HSCs were treated with acetaldehyde,and the expression of KDM3A mRNA was detected using real-time quantitative PCR(RT-PCR).Subsequently,HSCs were transfected with lentivirus to interfere with KDM3A expression for 72 h.After acetaldehyde stimulation,the HSCs were collected,and cell proliferation activity was assessed using the thiazolyl blue assay.The content of type Ⅲ and type Ⅰ collagen proteins was determined u-sing enzyme-linked immunosorbent assay,while the expression levels of transforming growth factor-beta 2(TGF-β2)were measured by RT-PCR and Western blotting.Results Compared with the control group,the relative expression levels of KDM3A mRNA in the acetaldehyde-treated group increased approximately 4-fold(1.00±0.10 vs.4.00±0.20),and TGF-β2 mRNA expression increased approximately 3-fold(1.00±0.40 vs.3.00±0.50),with both differences being statistically significant(t=43.201,44.032,P＜0.001).Following interference with KDM3A,the proliferation activity of transfected cells decreased by approximately 43.3％compared to the control group[transfected group optical density(0.38±0.06)lower than control group(0.67±0.08)].Compared to the control group,the content of type 111 collagen decreased by approximately 20.00％(1.00±0.10 vs.0.80±0.04),and type Ⅰ collagen content decreased by approximately 30.00％(1.00±0.10 vs.0.70±0.04),the protein and mRNA levels of TGF-β2 also significantly decreased(1.00±0.10 vs.0.70±0.10)in the transfected group,with all differences being statistically significant(t=12.272,12.621,21.369,P＜0.001).Conclusion Interfering with histone demethylase KDM3A can partially inhibit the proliferation and collagen synthesis of hepatic stellate cells,possibly by suppressing TGF-β2 expression.
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