Lee Tzung-Yan, Chang Hen-Hong, Wang Guei-Jane, Chiu Jen-Hwey, Yang Ying-Ying, Lin Han-Chieh
Graduate Institute of Traditional Chinese Medicine, Chang Gung University, Taoyuan, Taiwan.
J Pharm Pharmacol. 2006 May;58(5):659-65. doi: 10.1211/jpp.58.5.0011.
Apoptosis is one of the events that are involved in liver fibrogenesis. Thus, factors that affect apoptosis may be used to modulate liver fibrosis. We have recently reported that Salvia miltiorrhiza plays a protective role in carbon tetrachloride (CCl4)-induced hepatic fibrosis. In this study, we aimed to evaluate whether S. miltiorrhiza modulated CCl4-induced hepatic apoptosis in rats. Male Wistar rats were given orally either vehicle or water-extract of S. miltiorrhiza (50 mg kg(-1) twice daily) for nine weeks beginning from the start of CCl4 administration. A group of normal rats was included for comparison. Hepatocyte DNA fragmentation and cytosolic caspase-3 and caspase-8 activity were determined in the experimental animals. Hepatic cytosolic Bax, Bcl-2, cytochrome c, and calpain-mu expressions were measured by Western blot analysis. Hepatic mitochondrial glutathione levels were assessed by colorimetric assay. Compared with normal rats, rats receiving CCl4 alone showed profound DNA fragmentation associated with an increased cytosolic fraction of cytochrome c and calpain-mu protein expressions and a decreased mitochondrial glutathione level. In contrast, a decreased laddering of DNA fragmentation was noted in rats receiving CCl4 plus S. miltiorrhiza extract. The mitochondrial glutathione level was significantly increased in rats receiving CCl4 plus S. miltiorrhiza extract compared with those receiving CCl4 alone. Additionally, cytosolic caspase-3 activity and cytosolic fractions of Bax, Bcl-2, cytochrome c, and calpain-mu protein expressions were decreased in rats receiving CCl4 plus S. miltiorrhiza extract compared with those receiving CCl4 alone. The cytosolic caspase-8 activity in rats receiving CCl4 alone was no different from those receiving CCl4 plus S. miltiorrhiza extract. These results indicated that chronic administration of S. miltiorrhiza ameliorated CCl4-mediatd hepatic apoptosis in rats. This effect may be related to the antioxidant properties of S. miltiorrhiza.
细胞凋亡是参与肝纤维化形成的事件之一。因此，影响细胞凋亡的因素可用于调节肝纤维化。我们最近报道，丹参对四氯化碳（CCl4）诱导的肝纤维化具有保护作用。在本研究中，我们旨在评估丹参是否能调节CCl4诱导的大鼠肝细胞凋亡。从给予CCl4开始，雄性Wistar大鼠连续9周口服溶剂或丹参水提取物（50 mg·kg-1，每日两次）。纳入一组正常大鼠作为对照。测定实验动物的肝细胞DNA片段化以及胞质半胱天冬酶-3和半胱天冬酶-8活性。通过蛋白质印迹分析测量肝胞质中Bax、Bcl-2、细胞色素c和钙蛋白酶-μ的表达。通过比色法评估肝线粒体谷胱甘肽水平。与正常大鼠相比，单独接受CCl4的大鼠表现出明显的DNA片段化，伴有细胞色素c和钙蛋白酶-μ蛋白表达的胞质部分增加以及线粒体谷胱甘肽水平降低。相反，接受CCl4加丹参提取物的大鼠DNA片段化的梯状条带减少。与单独接受CCl4的大鼠相比，接受CCl4加丹参提取物的大鼠线粒体谷胱甘肽水平显著升高。此外，与单独接受CCl4的大鼠相比，接受CCl4加丹参提取物的大鼠胞质半胱天冬酶-3活性以及Bax、Bcl-2、细胞色素c和钙蛋白酶-μ蛋白表达的胞质部分降低。单独接受CCl4的大鼠的胞质半胱天冬酶-8活性与接受CCl4加丹参提取物的大鼠无差异。这些结果表明，长期给予丹参可改善CCl4介导的大鼠肝细胞凋亡。这种作用可能与丹参的抗氧化特性有关。
