目的探讨大果木姜子油（LLEO）对大鼠心肌缺血再灌注损伤（MIRI）的保护作用及其对心肌能量代谢的影响，并初步分析Ras同源家族成员A（RhoA）/Rho相关卷曲螺旋结构域蛋白激酶1（ROCK1）/ATP合酶δ亚基（ATP5D）轴相关调控变化。方法 SPF级雄性SD大鼠60只，随机分为假手术组、模型组、复方丹参滴丸（阳性药，0.4 g·kg^-1）组及LLEO低、中、高剂量（0.3、0.4、0.5 mL·kg^-1）组，每组10只，假手术组和模型组给予等体积溶媒（含1%聚山梨酯-80的0.9%氯化钠溶液），连续ig 14 d。末次给药后，采用左前降支冠状动脉结扎30 min、再灌注60 min建立MIRI模型，各组最终纳入分析大鼠8只。采用生物机能实验系统检测心功能，苏木素-伊红（HE）染色和透射电镜观察心肌病理及超微结构，酶联免疫吸附试验（ELISA）检测血清心肌损伤标志物，TUNEL染色检测心肌细胞凋亡，Western blotting检测凋亡相关蛋白及RhoA、ROCK1、ATP5D蛋白表达，比色法检测氧化应激指标及ATP合酶活性，试剂盒法检测三磷酸腺苷（ATP）、二磷酸腺苷（ADP）、一磷酸腺苷（AMP）水平并计算ATP/ADP、ATP/AMP值。结果与模型组比较，LLEO可改善心功能，减轻心肌病理及超微结构损伤，降低心肌损伤标志物水平、氧化应激水平和心肌细胞凋亡率，升高ATP/ADP、ATP/AMP值及ATP合酶活性，上调ATP5D表达，作用在中、高剂量组均差异显著（P＜0.05、0.01、0.001）；中、高剂量组可显著下调RhoA、ROCK1蛋白表达（P＜0.05、0.01）。结论 LLEO可减轻大鼠MIRI所致心功能障碍、心肌结构损伤、氧化应激和细胞凋亡，并改善心肌能量代谢，其作用可能与抑制RhoA/ROCK1异常激活、上调ATP5D表达并提高ATP合酶活性有关。

Objective To investigate the protective effects of Litsea lancilimba essential oil (LLEO) against myocardial ischemiareperfusion injury (MIRI) in rats and its effects on myocardial energy metabolism, and to preliminarily explore the regulatory changes related to the RhoA/ROCK1/ATP5D axis. Methods Sixty SPF-grade male SD rats were randomly divided into six groups: sham surgery group, model group, Compound Danshen Drip Pills group (positive control, 0.4 g·kg^-1), and LLEO low-, medium-, and high- dose groups (0.3, 0.4, and 0.5 mL·kg^-1), with 10 rats in each group. The sham and model groups received an equal volume of solvent (0.9% NaCl solution containing 1% polysorbate-80) by oral gavage for 14 consecutive days. After the final administration, myocardial ischemia-reperfusion injury (MIRI) was induced by ligating the left anterior descending coronary artery for 30 minutes followed by 60 minutes of reperfusion. Eight rats from each group were included in the final analysis. Cardiac function was assessed using a biological function experimental system; myocardial pathology and ultrastructure were examined via hematoxylin-eosin (HE) staining and transmission electron microscopy; serum myocardial injury markers were measured by ELISA; myocardial cell apoptosis was detected by TUNEL staining; expression of apoptosis-related proteins, RhoA, ROCK1, and ATP5D was analyzed by Western blotting; oxidative stress indicators and ATP synthase activity were determined by colorimetric assays; levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) were measured using commercial kits, and the ATP/ADP and ATP/AMP ratios were calculated. Results compared to the model group, LLEO improved cardiac function, reduced myocardial pathological and ultrastructural damage, decreased levels of myocardial injury markers and oxidative stress, and lowered the rate of myocardial cell apoptosis. It also increased the ATP/ADP and ATP/AMP ratios and ATP synthase activity, upregulated ATP5D expression, with significant differences observed in the medium- and high-dose groups (P < 0.05, 0.01, 0.001). Moreover, the medium- and high-dose groups significantly downregulated RhoA and ROCK1 protein expression (P < 0.05, 0.01). Conclusion LLEO can alleviate MIRIinduced cardiac dysfunction, myocardial structural injury, oxidative stress, and cardiomyocyte apoptosis, and improve myocardial energy metabolism in rats. Its effects may be associated with inhibition of abnormal RhoA/ROCK1 activation, upregulation of ATP5D expression, and enhancement of ATP synthase activity.

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国家自然科学基金资助项目（82060770）；贵州中医药大学国家与省级科技创新人才团队培育项目（贵中医[2024]002号）