Ze Xingyu, Jia Jidong, Li Xinmin, You Hong, Zhao Xinyan, Zhang Dong, Wang Baoen
Liver Disease Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, P.R. China.
Beijing Institute of Integrated Traditional and Western Medicine, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, P.R. China.
Mol Med Rep. 2016 Feb;13(2):1501-8. doi: 10.3892/mmr.2015.4696. Epub 2015 Dec 18.
Tanshinone IIA (TSA) is a widely used traditional Chinese medicine, which has been demonstrated to protect damaged liver cells and is currently administered in the treatment of liver fibrosis. Liver precursor cells, also termed oval cells, are key in the repair of liver tissues following injury. However, whether TSA improves the function of liver cells and protects the liver from injury by enhancing the growth and proliferation of hepatic oval cells remains to be elucidated. In the present study, low to moderate concentrations of TSA were observed to stimulate proliferation, did not induce apoptosis in WB-F344 rat hepatic oval cells and the increased expression levels of β-catenin. WB-F344 cells were treated with various concentrations of TSA (0-80 µg/ml) for 24, 48, 72 and 96 h. Cell proliferation was measured using a Cell Counting kit-8 (CCK-8) assay, a 5-ethynyl-2'-deoxyuridine assay and a carboxyfluorescein diacetate succinimidyl ester (CFSE) assay. The CCK-8 assay demonstrated that treatment of WB-F344 cells with 20-40 µg/ml TSA for up to 72 h significantly increased proliferation. Similar results were observed in the subsequent EdU and CFSE assays. Furthermore, a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay demonstrated that 20-40 µg/ml TSA treatment for up to 96 h did not induce apoptosis of the WB-F344 cells. Notably, the results of western blot, immunofluorescence and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that treatment of the WB-F344 cells with 20-40 µg/ml TSA for up to 72 h significantly increased the expression levels of β-catenin. These data indicated that TSA at concentrations between 20 and 40 µg/ml may induce WB-F344 cell proliferation by activating the canonical Wnt signaling pathway. The results of the present study suggest that TSA may be a useful natural agent to enhance repair and regeneration of the injured liver, and improve liver regeneration following orthotopic liver transplantation.
丹参酮IIA（TSA）是一种广泛应用的传统中药，已被证明可保护受损肝细胞，目前用于治疗肝纤维化。肝前体细胞，也称为卵圆细胞，是肝组织损伤后修复的关键。然而，TSA是否通过增强肝卵圆细胞的生长和增殖来改善肝细胞功能并保护肝脏免受损伤仍有待阐明。在本研究中，观察到低至中等浓度的TSA可刺激增殖，不会诱导WB-F344大鼠肝卵圆细胞凋亡，且β-连环蛋白表达水平增加。用不同浓度的TSA（0-80μg/ml）处理WB-F344细胞24、48、72和96小时。使用细胞计数试剂盒-8（CCK-8）检测、5-乙炔基-2'-脱氧尿苷检测和羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）检测来测量细胞增殖。CCK-8检测表明，用20-40μg/ml TSA处理WB-F344细胞长达72小时可显著增加增殖。在随后的EdU和CFSE检测中观察到类似结果。此外，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记检测表明，用20-40μg/ml TSA处理长达96小时不会诱导WB-F344细胞凋亡。值得注意的是，蛋白质免疫印迹、免疫荧光和逆转录-定量聚合酶链反应分析结果表明，用20-40μg/ml TSA处理WB-F344细胞长达72小时可显著增加β-连环蛋白的表达水平。这些数据表明，浓度在20至40μg/ml之间的TSA可能通过激活经典Wnt信号通路诱导WB-F344细胞增殖。本研究结果表明，TSA可能是一种有用的天然药物，可增强受损肝脏的修复和再生，并改善原位肝移植后的肝脏再生。
