Abstract: Objective　To compare the effects of levo⁃praziquantel (L⁃PZQ) and dextro⁃praziquantel (D⁃PZQ) on the proliferation and activation of the human hepatic stellate cell line LX⁃2 in vitro. Methods　LX⁃2 cells were stimulated with transforming growth factor⁃β (TGF⁃β). LX⁃2 cell proliferation was measured using the CCK⁃8 assay after 24 h stimulation with 0 to 50 μg/mL concentrations of praziquantel, and the gene and protein expression of typeⅠcollagen (collagenⅠ), type Ⅲ collagen (collagen Ⅲ) and α⁃smooth muscle actin (α⁃SMA) was quantified in LX⁃2 cells using quantitative real⁃time PCR (qPCR) and Western blotting assays 24 h and 48 h following stimulation with 15 μg/mL praziquantel to detect LX⁃2 cell activation. Results　There were significant differences in the survival rate of LX⁃2 cells between L⁃PZQ and D⁃PZQ treatments at all concentrations (F = 6.119 and 79.180, both P values < 0.05). Either L⁃PZQ or D⁃PZQ at a concentration of < 30 μg/mL showed no remarkable effects on the LX⁃2 cell proliferation (both P values > 0.05), and L⁃PZQ at a concentration of > 50 μg/mL and D⁃PZQ at a concentration of > 40 μg/mL inhibited the LX⁃2 cell proliferation (both P values < 0.05), while D⁃PZQ at concentrations of 40 μg/mL and 50 μg/mL showed greater inhibition on LX⁃2 cell proliferation than L⁃PZQ (t = 3.419 and 8.776, both P values < 0.05). There were significant differences in the collagenⅠ, collagen Ⅲ and α⁃SMA expression in LX⁃2 cells at both transcriptional (F = 21.55, 79.99 and 46.70, all P values < 0.05) and translational levels (F = 20.12, 30.29 and 32.93, all P values < 0.05) among the blank control group, TGF⁃β stimulation group, L⁃PZQ treatment group and D⁃PZQ treatment group. L⁃PZQ treatment resulted in remarkable inhibition on collagen Ⅲ and α⁃SMA gene expression in LX⁃2 cells (both P values < 0.05); however, the treatment showed no remarkable inhibition collagenⅠ gene expression or collagenⅠ, collagen Ⅲ or α⁃SMA protein expression in LX⁃2 cells (all P values > 0.05). In addition, D⁃PZQ treatment resulted in significant inhibition on collagenⅠ, collagen Ⅲ and α⁃SMA expression in LX⁃2 cells at both translational and transcriptional levels (all P values < 0.05), and D⁃PZQ showed higher inhibition on collagenⅠ, collagen Ⅲ and α⁃SMA gene expression in LX⁃2 cells than L⁃PZQ (all P values < 0.05). Conclusions　Both L⁃PZQ and D⁃PZQ inhibit the proliferation and activation of LX⁃2 cells, and D⁃PZQ shows a higher inhibitory activity than L⁃PZQ.

Key words: Praziquantel, Isomer, Hepatic stellate cell, Proliferation, Activation

摘要： 目的　观察左旋吡喹酮（L⁃PZQ）和右旋吡喹酮（D⁃PZQ）体外对人肝星状LX⁃2细胞系增殖及活化的作用差异。方法　利用转化生长因子⁃β（TGF⁃β）刺激激活LX⁃2细胞；采用CCK⁃8法检测以0 ~ 50 μg/mL梯度浓度吡喹酮刺激24 h后的LX⁃2细胞增殖情况；采用实时荧光定量PCR（qPCR）和免疫印迹试验检测15 mg/mL吡喹酮刺激LX⁃2细胞24 h 后的Ⅰ型胶原、Ⅲ型胶原和α⁃平滑肌肌动蛋白（α⁃SMA）基因表达水平和48 h后蛋白表达水平，以分析LX⁃2细胞活化情况。结果　L⁃PZQ和D⁃PZQ两药物各浓度组LX⁃2细胞存活率差异均有统计学意义（F = 6.119、79.180，P均< 0.05）；药物浓度< 30 μg/mL时，L⁃PZQ和D⁃PZQ对激活型LX⁃2增殖能力均无显著影响（P均> 0.05）；L⁃PZQ浓度> 50 μg/mL和D⁃PZQ浓度> 40 μg/mL时，均对激活型LX⁃2增殖能力产生抑制作用（P均< 0.05）；D⁃PZQ浓度为40 μg/mL和50 μg/mL时，对激活型LX⁃2增殖的抑制作用均强于L⁃PZQ（t = 3.419、8.776，P均< 0.05）。空白组、TGF⁃β诱导组、L⁃PZQ组和D⁃PZQ组LX⁃2细胞中Ⅰ型胶原、Ⅲ型胶原和α⁃SMA基因（F = 21.55、79.99、46.70，P 均< 0.05）和蛋白表达水平（F = 20.12、30.29、32.93，P 均< 0.05）差异均有统计学意义。L⁃PZQ对激活型LX⁃2细胞中Ⅲ型胶原和α⁃SMA基因表达水平具有显著抑制作用（P均< 0.05），但对Ⅰ型胶原基因表达水平及Ⅰ型胶原、Ⅲ型胶原和α⁃SMA蛋白表达水平均无显著影响（P均> 0.05）；D⁃PZQ对激活型LX⁃2细胞中Ⅰ型胶原、Ⅲ型胶原和α⁃SMA基因和蛋白表达水平均有显著抑制作用（P均< 0.05）；D⁃PZQ对激活型LX⁃2细胞中Ⅰ型胶原、Ⅲ型胶原和α⁃SMA基因表达抑制作用强于L⁃PZQ（P均< 0.05）。结论　L⁃PZQ和D⁃PZQ对LX⁃2细胞增殖及活化均有一定抑制作用，且D⁃PZQ的抑制作用强于L⁃PZQ。

关键词: 吡喹酮, 异构体, 肝星状细胞, 增殖, 活化