摘要
背景:淫羊藿苷可用于心血管疾病的治疗,但是由于淫羊藿苷的脂溶性高、水溶性差、肝脏首过效应明显,导致其生物利用度较低,限制了其临床应用。目的:将淫羊藿苷负载于壳聚糖/胶原复合水凝胶中,观察其治疗大鼠心肌梗死的效果。方法:制备负载淫羊藿苷不同量(0.4,0.8,1.6 mg/mL)的壳聚糖/胶原复合水凝胶,分别记为CS/COI/ICA0.4、CS/COI/ICA0.8、CS/COI/ICA1.6,检测水凝胶的细胞相容性与体外缓释性能。取成年SD大鼠,结扎左冠状前支建立心肌梗死模型,将造模成功的96只大鼠随机4组治疗,模型对照组梗死区域、梗死区周围及后壁区域注射PBS,CS/COI/ICA0.4组、CS/COI/ICA0.8组、CS/COI/ICA1.6组梗死区域、梗死区周围及后壁区域分别注射对应的水凝胶,每组24只;同时选取造模外的24只大鼠作为正常对照。治疗28 d后检测相关指标。结果与结论:①体外细胞实验:负载淫羊藿苷不同量的壳聚糖/胶原复合水凝胶具有良好的细胞相容性,有利于心肌细胞的增殖与存活;3种负载量水凝胶均可持续释放淫羊藿苷达32 d以上。②动物实验:与正常对照组相比,模型对照组大鼠心脏结构发生严重重塑、心功能显著下降(P<0.01);与模型对照组比较,各CS/COI/ICA组大鼠心脏结构、心功能均有明显改善(P<0.05),其中以CS/COI/ICA1.6组改善最明显(P<0.05)。Masson三色染色显示,各CS/COI/ICA组大鼠心肌组织纤维化程度显著轻于模型对照组(P<0.05),心室壁厚度大于模型对照组(P<0.05)。免疫组化染色显示,模型对照组大鼠心肌组织CD31、α-平滑肌肌动蛋白含量低于正常对照组(P<0.05),各CS/COI/ICA组大鼠心肌组织CD31、α-平滑肌肌动蛋白含量高于模型对照组(P<0.05)。RT-PCR检测显示,模型对照组心肌组织血管内皮生长因子、碱性成纤维细胞生长因子的mRNA表达量低于正常对照组(P<0.05),各CS/COI/ICA组血管内皮生长因子、碱性成纤维细胞生长因子的mRNA表达量高于模型对照组(P<0.05)。③结果表明,壳聚糖/胶原/淫羊藿苷复合水凝胶可能通过提升梗死部位血管内皮生长因子与碱性成纤维细胞生长因子的表达刺激血管新生,进而改善心肌组织有效灌注量不足的局部微环境,达到有效抑制心室重塑、改善心功能的治疗作用。
BACKGROUND:Icariin can be used in the treatment of cardiovascular diseases,however,its high lipid solubility,poor water solubility,and obvious hepatic firstpass effect result in low bioavailability,which limits its clinical application.OBJECTIVE:To observe the effects of icariin combined with injectable chitosan/collagen composite hydrogel on myocardial infarction in rats.METHODS:Chitosan/collagen composite hydrogels loaded with different concentrations(0.4,0.8,and 1.6 mg/mL)of icariin were prepared and marked as CS/COI/ICA0.4,CS/COI/ICA0.8,and CS/COI/ICA1.6,respectively.The cytocompatibility and sustained release performance of the hydrogels were tested.The le?t anterior descending artery of adult SD rats was ligated to establish a myocardial infarction model.Ninety-six myocardial infarction rat models were randomly divided into four groups(24 rats in each group):model,CS/COI/ICA0.4,CS/COI/ICA0.8,and CS/COI/ICA1.6 groups,and the infarct area,peri-infarct area and posterior wall area of rats in these four groups were injected with PBS or hydrogels.Normal rats were selected as normal controls.The experimental parameters were measured a?ter 28 days of treatment.RESULTS AND CONCLUSION:(1)In vitro cell experiments showed that chitosan/collagen composite hydrogels loaded with different concentrations of icariin had good cytocompatibility,which was beneficial to the proliferation and survival of cardiomyocytes.The release of icariin from all the three hydrogels kept for more than 32 days.(2)The animal experiments showed that the cardiac structure of rats in the model group was severely remodeled and the cardiac function was significantly decreased compared with the normal control group(P<0.01).The CS/COI/ICA0.4,CS/COI/ICA0.8,and CS/COI/ICA1.6 groups had a significant improvement in cardiac structure and cardiac function compared with the model control group(P<0.05),with the most significant improvement in the CS/COI/ICA1.6 group(P<0.05).Masson staining showed that the degree of myocardial fibrosis in the CS/COI/ICA0.4,CS/COI/ICA0.8,and CS/COI/ICA1.6 groups was significantly less(P<0.05),while the thickness of ventricular wall was significantly greater compared with that in the model group(P<0.05).Immunohistochemical staining and quantitative reverse transcription-polymerase chain reaction showed that the immunoreactivities of CD31 andα-smooth muscle actin and mRNA expression of vascular endothelial growth factor and basic fibroblast growth factor were significantly decreased compared with the normal control group(all P<0.05),while those were significantly increased in the CS/COI/ICA0.4,CS/COI/ICA0.8,and CS/COI/ICA1.6 groups compared with the model group(all P<0.05).(3)These findings suggest that chitosan/collagen/icaricin composite hydrogels can stimulate angiogenesis by increasing the expression of vascular endothelial growth factor and basic fibroblast growth factor in myocardial tissues in infracted areas to improve the local microenvironment of myocardial tissue due to insufficient perfusion,thereby inhibiting ventricular remodeling and improving cardiac function.
作者
何文凤
薛成
郑健康
帅壮
岳荣川
He Wenfeng;Xue Cheng;Zheng Jiankang;Shuai Zhuang;Yue Rongchuan(Department of Cardiology,Affiliated Hospital of North Sichuan Medical University,Cardiovascular Disease Laboratory,Nanchong 637000,Sichuan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第25期3992-3998,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(81600232),项目名称:Calpain调控内质网应激对NLRP3的影响在心肌缺血再灌注细胞焦亡损伤中的作用机制研究,项目负责人:岳荣川
医学院校级重点项目(CBY18-A-ZD14),项目名称:苯磺酸氨氯地平对人外周血巨噬细胞吞噬脂质作用的研究,项目负责人:何文凤。
