Page 58 - 中国全科医学2022-17

http://www.chinagp.net E-mail:zgqkyx@chinagp.net.cn ·2103·

widely used in the treatment of various tumors，especially solid tumors. However，its strong cardiotoxicity-induced irreversible
cardiomyopathy and congestive heart failure limit its clinical application. Yellow Wine Polyphenols Compounds（YWPC）
are polyphenols extracted from Shaoxing yellow rice wine which have anti-inflammatory and antioxidant activities，and can
alleviate DOX-induced myocardial injury，but the mechanism of actions is still unclear. Objective To explore the mechanism
of YWPC reducing DOX-induced myocardial injury via in vitro and in vivo experiments. Methods SD rats were used for an in
vivo experiment and divided into four groups：control group（intervened with normal saline），YWPC group（intervened with
YWPC and normal saline），DOX group（intervened with DOX and normal saline for establishing a DOX-induced myocardial
injury model） and DOX+YWPC group（intervened with DOX，YWPC and normal saline for observing the myocardial protective
effect of YWPC in a DOX-induced myocardial injury model）. When the experiment ended，all rats were sacrificed and cardiac
tissues were taken out for examining myocardial fiber morphology using Masson's trichrome staining，myocardial cell apoptosis
using TUNEL assay，pathological changes using immunohistochemical assay，and levels of proteins（Bcl-2 and Bax） involved
in apoptosis as well as expression level of SIRT3 using Western blotting. And the serum lactate dehydrogenase（LDH） was also
measured. Cardiomyocyte H9C2 cells of rats were used for an in vitro experiment and divided into five groups：control group，
YWPC group（intervened with YWPC and normal saline），DOX group（intervened with DOX for establishing a DOX-
induced myocardial injury model within 24 hours） and three DOX+YWPC（1 mg/L，10 mg/L，100 mg/L） groups〔first
intervened with DOX for establishing a DOX-induced myocardial injury model within 24 hours，then with YWPC for observing
the myocardial protective effect of three concentrations of YWPC（1 mg/L，10 mg/L，100 mg/L），respectively〕. Viability of
H9C2 cells was measured by CCK-8 assay. Apoptosis，and SIRT3 expressed in H9C2 cells were measured by Western blotting.
Then another batch of the same H9C2 cells were took and divided into control group，DOX group，DOX+YWPC group，
DOX+3-TYP group and DOX+YWPC+3-TYP group，and SIRT3 inhibitor 3-TYP was used to inhibit the activity of SIRT3
protein in the latter two groups. Then apoptosis level and SIRT3 protein expressed in H9C2 cells were detected by Western blotting
for further assessing the effect of SIRT3 in reducing DOX-induced myocardial injury. Results In vivo experiment：（1）
Under the microscope，Masson's trichrome-stained myocardial fibers of rats in DOX group were disordered and intersected with
a large number of blue collagen fibers. The myocardial texture of DOX+YWPC group was partially restored with decreased blue
collagen fibers. DOX group had higher proportion of collagen fibers distributed in cardiovascular tissues，and higher serum LDH
than control and DOX+YWPC groups（P<0.05）. DOX group had lower ratio of Bcl-2/Bax and level of expression of SIRT3 than
control and DOX+YWPC groups（P<0.05）. The apoptotic myocardial cells with a patch distribution appearing as bright green
dots were significantly increased in DOX group at first，but were decreased after YWPC treatment. In vitro experiment：（1）
The absorbance value，Bcl-2/Bax ratio and expression level of SIRT3 in DOX group were lower than those of control group，
DOX+YWPC（1 mg/L） group，DOX+YWPC（10 mg/L） group and DOX+YWPC（100 mg/L） group（P<0.05）.（2）The
Bcl-2 /Bax ratio and expression level of SIRT3 in DOX or DOX+3-TYP group were lower than those in control group（P<0.05）.
（3）The Bcl-2/Bax ratio in DOX+YWPC+3-TYP group was lower than that in DOX+YWPC group（P<0.05）.（4）The
expression level of SIRT3 was similar in DOX+YWPC and DOX+YWPC+3-TYP groups（P>0.05）. Conclusion DOX-induced
myocardial injury in in vitro and in vivo experiments with rats may be alleviated by YWPC via improving the expression level of
SIRT3，and the effect may be reduced if the expression level of SIRT3 protein is inhibited.
【Key words】 Heart diseases；Doxorubicin；Yellow Wine Polyphenol Compounds；Antineoplastic agents；SIRT3；
Apoptosis

自 20 世纪 50 年代第一个蒽环类药物被发现以来， 自由基和保护心血管作用的天然化学物，本研究团队
因其在各类肿瘤治疗中的明显效果而被广泛应用于临床 前期研究以黄酒及其主要成分黄酒多酚（Yellow Wine
治疗 ［1］ 。阿霉素（即多柔比星，Doxorubicin，DOX） Polyphenol Compounds，YWPC）为研究对象，探究其
是一种蒽环类药物，对白血病及多种实体肿瘤均有较好 对心血管系统的保护作用，结果发现 YWPC 具有缓解
的临床治疗效果，但在临床应用中常因其剂量依赖性、 DOX 所致心脏毒性的作用。研究表明，YWPC 可以通
进展性、不可逆性心肌损伤而受到限制，如 DOX 在儿 过调节细胞内核因子 E2 相关因子 2（Nrf2）的核易位水
童癌症治疗后会形成长期心肌损伤 ［2-3］ 。DOX 诱导的 平缓解 DOX 引起的氧化应激及心脏毒性 ［5-6］ 。除引起
心肌损伤机制十分复杂，包括线粒体功能障碍、DNA 氧化应激反应外，破坏线粒体功能的完整性也是 DOX
损伤、铁代谢受损、细胞凋亡及自噬失调等多种途径 ［4］ ， 引起心脏毒性的重要原因，而沉默调节蛋白 3（SIRT3）
目前如何预防蒽环类药物引起的心肌损伤仍无法形成共 是线粒体中的Ⅲ类组蛋白脱乙酰基酶，主要通过调节赖
识。多酚类物质是一类具有抗肿瘤、抗氧化、清除氧 氨酸的乙酰化来调节线粒体网络。SIRT3 通过对细胞线