CN115025106A - 人参皂苷Rk3在制备具有神经保护作用药物中的用途 - Google Patents
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Abstract
本发明公开了人参皂苷Rk3在制备具有神经保护作用药物中的用途，本发明中人参皂苷Rk3具有显著的神经保护活性，能够显著提高H2O2诱导的神经损伤SHSY‑5Y细胞的存活率，对SHSY‑5Y神经细胞中ROS产生具有显著抑制效果；人参皂苷Rk3能够用于制备新的神经保护药物，用于改善和治疗抑郁症、焦虑症以及神经退行性疾病等神经病症，本发明开拓了人参皂苷Rk3的应用范围，为治疗神经疾病提供了新途径。
Description
技术领域
本发明属生物医药技术领域，具体涉及人参皂苷Rk3在神经保护方面中的应用。
背景技术
神经疾病是指由慢性进行性的中枢神经组织退行性变性而产生的疾病的总称，例如运动失调的帕金森疾病（PD）、老年性痴呆的阿尔茨海默病（AD）、亨廷顿病（HD）、肌萎缩侧索硬化症（ALS）等疾病。目前临床上常用治疗神经疾病的药物，主要包括营养神经类药物，如甲钴胺胶囊等，常用于神经损伤的修复治疗。改善脑代谢，促进微循环的药物主要有丁苯酞胶囊、丹参川芎嗪注射液等，以调节脑血流供应。改善大脑认知状态的药物主要包括奥拉西坦胶囊等，能够改善患者健忘症状，这些药物均有诸多不良反应。近年研究发现，稀有人参皂苷具有多种生物活性，可能作为治疗神经疾病的潜在药物。
人参皂苷Rk3为稀有皂苷，通常通过常见人参皂苷转化而得。稀有皂苷因为比普通皂苷具有更强的生物活性而备受关注。据报道，人参皂苷Rk3和含有人参皂苷Rk3的药物组合可用于治疗肿瘤（发明专利CN201610616139.8）、毛细血管扩张症（发明专利CN201610607005.X）、血管新生疾病（发明专利CN 201610605891.2）、酒精性脂肪肝（发明专利CN 202010305247.X）、抗光老化（发明专利CN 202010419961.1）等，但是对于人参皂苷Rk3的神经保护作用尚未有报道。
发明内容
本发明提供了一种人参皂苷Rk3的新用途，即其在制备具有神经保护作用药物中的应用。
本发明中所述人参皂苷Rk3的化学结构式如下：
本发明中人参皂苷Rk3具有显著的神经保护活性，能够显著提高H2O2诱导的神经损伤SHSY-5Y细胞的存活率，对SHSY-5Y神经细胞中ROS产生显著抑制效果。
本发明所述的应用是以人参皂苷Rk3作为制备具有神经保护作用药物的活性成分，还可以加入一种或多种药物上可接受的辅料，以改善药物吸收效果或便于服用，如制成胶囊或丸剂、粉剂、片剂、粒剂、口服液、注射液、气雾剂等，即制成药剂学上适宜的使用剂型。
本发明人参皂苷Rk3还可以与其他具有神经保护作用的活性成分的组合作为药物的活性成分。
本发明提供了人参皂苷Rk3在制备神经保护药物中应用，神经疾病包括抑郁症、焦虑症以及神经退行性疾病等。
本发明为人参皂苷Rk3的医疗用途提供了一个新的研究领域，本发明人参皂苷Rk3的神经保护活性进行了研究，实验结果显示人参皂苷Rk3能够显著促进神经细胞生长，提高细胞存活率，保护细胞免于ROS的损伤，人参皂苷Rk3具有神经保护作用，能够用于治疗或改善神经疾病。
附图说明
图 1 人参皂苷Rk3对H2O2诱导SHSY-5Y细胞的影响。
具体实施方式
下面结合附图和实施例对本发明作进一步详细说明，但本发明保护范围不局限于所述内容，实施例中使用的试剂，如无特殊说明，均为常规市售试剂或按常规方法配制的试剂，实施例中使用方法，如无特殊说明均为常规实验方法。
实施例1：人参皂苷Rk3对H2O2诱导神经母瘤细胞SHSY-5Y细胞存活率的影响
将处于对数生长期的SH-SY5Y细胞稀释到2×105个/mL的浓度后，接种于96孔板中，每孔100μL，37℃孵育24h后，空白对照组加入含10%胎牛血清的DMEM/F12完全培养液，模型组和给药组分别加入含300μmo1/L H2O2的DMEM/F12完全培养液，37℃条件下孵育1 h后，将浓度10、20、30μmol/L的人参皂苷Rk3、30μmol/L其他人参皂苷加入不同给药组中，孵育24h后，各组除去上清，再加入DMEM/F12完全培养液后，每孔加入10μL CCK8，37℃孵育2h后，通过酶标仪于450nm波长下，测量每孔吸光度值；每个实验组3个重复，空白对照组的吸光度值设定为1，其余组吸光度值/空白对照组实际值来表示各种细胞存活率；
结果见表1、图1，由表1可知，与空白对照组相比，H2O2诱导SHSY-5Y细胞生存率显著降低（p<0.001）；与模型组相比，人参皂苷Rk3能够呈浓度依赖性地提高SHSY-5Y细胞生存率（p<0.001）。从图1可以看出，人参皂苷Rk3对H2O2诱导SHSY-5Y细胞损伤具有保护作用，存活的细胞多于模型组；与其他人参皂苷化合物相比，人参皂苷Rk3提高SHSY-5Y细胞生存率的作用最强。
表1 人参皂苷Rk3提高H2O2诱导神经母瘤细胞SHSY-5Y的细胞存活率(
，n=3)
注：与空白组相比，### p<0.001；与模型组相比，*** p<0.001。
实施例2：人参皂苷Rk3对H2O2诱导SHSY-5Y细胞内ROS水平影响
将处于对数生长期的SH-SY5Y细胞稀释到2×105个/mL的浓度后，接种于96孔板中，设置空白对照组、模型组、加药组，每个组3个平行，每孔100μL，37℃孵育24h后，加药组中加入不同类型人参皂苷（30μmol/L）后培养24 h，各组除去培养液，再加入含300 mo1/LH2O2的DMEM/F12完全培养液，培养2h；再次除去含H2O2的培养液，加入含5μmol/L DCFH-DA活性氧荧光探针的PBS缓冲液（pH=7.4），在37℃下孵育30min后；用PBS缓冲液洗净未进入细胞的荧光探针，于酶标仪（激发波长485nm，发射波长525nm）上检测荧光强度；
结果见表2，由表可知，与空白对照组相比，模型组H2O2诱导SHSY-5Y细胞内ROS水平升高（p<0.001）；与模型组相比，人参皂苷Rk3能够降低SHSY-5Y细胞内ROS水平（p<0.001）。从细胞内ROS水平角度，进一步验证人参皂苷Rk3对H2O2诱导SHSY-5Y细胞损伤的保护作用；与其他人参皂苷化合物相比，人参皂苷Rk3抑制SHSY-5Y细胞产生ROS的作用最强。
表 2 人参皂苷Rk3抑制H2O2诱导神经母瘤细胞SHSY-5Y ROS的产生(
，n=3)
注：与空白组相比，### p<0.001；与模型组相比，*** p<0.001。
Claims (2)
1.人参皂苷Rk3在制备具有神经保护作用药物中的应用。
2.根据权利要求1所述的应用，其特征在于：具有神经保护作用药物包括治疗抑郁症、焦虑症、神经退行性疾病的药物。
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