大果木姜子油的提取工艺优化及其对心肌缺血改善作用的研究

目的 采用响应面法优化大果木姜子油的提取工艺, 并探讨不同来源大果木姜子油对心肌缺血的改善作用。方法 采用单因素实验法考察提取溶剂、液料比、物料粒径、提取温度和提取时间5个单因素对大果木姜子油得率的影响, 并在此基础上运用响应面法就液料比、物料粒径和提取温度3个因素进行大果木姜子油提取工艺的优化。建立大鼠心肌缺血模型, 采用免疫印迹法检测大鼠心肌组织中兔抗人单克隆抗体(Bcl-2 assaciated xprotein, Bax)及B淋巴细胞瘤-2基因(B-cell lymphoma-2, Bcl-2)蛋白的表达、采用聚合酶链式反应检测心肌组织中碱性成纤维生长因子(fibroblast growth factor 2, FGF-2)的表达、采用免疫组化法检测心肌组织中纤溶酶原激活物抑制剂-1 (plasminogen activator inhibitor-1, PAI-1)及组织型纤溶酶原激活剂(tissue-type plasminogenactivator, TPA)的表达, 并采用苏木精-伊红染色法观察心肌组织病理变化。结果 经响应面法分析大果木姜子油的最佳提取工艺为: 液料比7:1 (mL:g), 粒径过80目, 提取温度90℃, 得率为51.23%。药理实验表明: 与空白组相比, 模型组中Bcl-2、FGF-2和TPA的水平下降, 而Bax和PAI-1水平上升, 经大果木姜子油治疗后Bax、PAI-1的水平下降, 而FGF-2、TPA和Bcl-2水平上升。结论 响应面法优化后的大果木姜子油的提取工艺条件稳定、可行, 且大果木姜子油具有改善心肌缺血的作用。本研究为提取和应用大果木姜子油提供了坚实的理论基础。

Optimization of the extraction process of Cinnamomum migao oil and its ameliorative effect on myocardial ischemia

Objective To optimize the extraction process of Cinnamomum migao oil using response surface methodology and explore the improvement effect of different sources of Cinnamomum migao oil on myocardial ischemia. Methods A single factor experimental method was used to investigate the effects of five single factors, namely extraction solvent, liquid-solid ratio, material particle size, extraction temperature, and extraction time, on the yield of Cinnamomum migao oil. Based on this, response surface methodology was used to optimize the extraction process of Cinnamomum migao oil using three factors: liquid-solid ratio, material particle size, and extraction temperature. Establishing a rat myocardial ischemia model, Western blot (WB) was used to detect the expression of Bcl-2 assaciated xprotein (Bax) and B-cell lymphoma-2 (Bcl-2) protein in rat myocardial tissue, Polymerase chain reaction (PCR) was used to detect the expression of Fibroblast growth factor 2 (FGF-2) in myocardial tissue, and immunohistochemistry (IHC) was used to detect the expression of Plasminogen activator inhibitor-1 (PAI-1) and Tissue-type plasminogenactivator (TPA) in myocardial tissue, Hematoxylin eosin staining (HE) was used to observe the pathological changes of myocardial tissue. Results The optimal extraction process for Cinnamomum migao oil was as follows: liquid-solid ratio 7:1 (mL/g), particle size over 80 mesh, extraction temperature 90℃, and yield of 51.23%. Pharmacological experiments had shown that compared with the blank group, the levels of Bcl-2, FGF-2, and TPA in the model group decreased, while the levels of Bax and PAI-1 increased. After treatment with Cinnamomum migao oil, the levels of Bax and PAI-1 decreased, while the levels of FGF-2, TPA, and Bcl-2 increased. Conclusion The extraction process conditions of Cinnamomum migao oil optimized by response surface methodology were stable and feasible, and Cinnamomum migao oil has the effect of improving myocardial ischemia. This study provides a solid theoretical basis for the extraction and application of Cinnamomum migao oil.