CN114632086A - 野黄芩苷增强碘-125粒子放射敏感性的应用 - Google Patents
野黄芩苷增强碘-125粒子放射敏感性的应用 Download PDFInfo
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Abstract
本发明属于生物医药领域，提供了一种野黄芩苷作为125I放射治疗恶性肿瘤增敏剂的用途。本发明提供了野黄芩苷作为125I放射治疗增敏剂的用途，通过试验验证了相比于125I放射性粒子单一作用，野黄芩苷与125I放射性粒子联合作用能够有效增强对靶细胞增殖的抑制，增强其促进细胞凋亡的能力。在临床应用中，对于存在125I放射抵抗的患者，联合使用野黄芩苷可以增强125I的治疗效果；并且125I放射治疗增敏剂的使用，可以降低125I的使用活度和临床治疗剂量，从而降低放射治疗相关并发症的发生率，从而提高患者治疗效果和生存质量。
Description
技术领域
本发明属于生物医药领域，具体涉及野黄芩苷增强肝细胞癌对125I放射性粒子的辐射敏感性的新应用。
背景技术
125I放射性粒子植入技术的使用和推广，提高了晚期肝细胞癌临床治疗的安全性和有效性，尤其是与其他化疗药物（如洛铂）和中药制剂联合使用时，预后明显改善。然而，125I放射性粒子对肝细胞癌细胞作用的具体机制仍有待研究。此外，中药制剂是否能增强肝细胞癌对125I放射性粒子的辐射敏感性尚不清楚，探讨125I放射性粒子的作用机制和新靶点，将有助于125I放射性粒子更好地应用于临床，并为肝细胞癌提供新的治疗思路。
近年来，天然药物的抗肿瘤作用受到越来越多的关注。科技的进步使中医辨证从方剂深入中药有效成分的探究，深入到细胞、基因层面。许多传统中药具有强大的抗肿瘤作用，可显著抑制肿瘤生长，且副作用较小。野黄芩苷(Scutellarin, SCU)是一种黄酮类化合物，又名灯盏花乙素，可从黄芩、半枝莲、白花蛇舌草等传统中药内提取，是其有效作用成分。SCU在多项实验研究中被证实对包括胃癌、肠癌、肺癌在内的多种肿瘤细胞具有显著的抑制作用，其可诱导肿瘤细胞凋亡、抑制肿瘤细胞增殖，增加化疗药物的敏感性，但其是否可能增加放疗的敏感性，特别是增加碘125粒子治疗肝癌的疗效，二者通过何种途径起作用，具体机制目前尚不清楚。
发明内容
针对现有技术中的问题，本发明提供一种野黄芩苷的新应用，能够增强肝细胞癌对125I放射性粒子的辐射敏感性。
为实现上述目的，本发明采用如下技术方案。
一种野黄芩苷作为125I放射治疗恶性肿瘤增敏剂的应用。
所述恶性肿瘤选自胃癌、肠癌、肺癌或肝癌。
本发明具有以下优点：
本发明提供了野黄芩苷作为125I放射治疗增敏剂的用途，通过试验验证了相比于125I放射性粒子单一作用，野黄芩苷与125I放射性粒子联合作用能够有效增强对靶细胞增殖的抑制，增强其促进细胞凋亡的能力。在临床应用中，对于存在125I放射抵抗的患者，联合使用野黄芩苷可以增强125I的治疗效果；并且125I放射治疗增敏剂的使用，可以降低125I的使用活度和临床治疗剂量，从而降低放射治疗相关并发症的发生率。从而提高患者治疗效果和生存质量。
附图说明
图1野黄芩苷浓度对数-HCC细胞抑制率曲线；
图2为不同处理对细胞周期的影响；
图3为不同处理对细胞凋亡的影响；
图4为不同处理对细胞增殖的影响；
其中，*表示p<0.05，**表示p<0.01，***表示p<0.001。
具体实施方式
下面结合实施例和附图对本发明做进一步说明，但本发明不受下述实施例的限制。
实施例1 野黄芩苷对125I的体外增敏试验
肝细胞癌细胞系SMMC7721和HepG2购自中乔新舟生物技术公司。SMMC7721细胞在RPMI 1640（Corning, Inc.）中培养，补充有10%胎牛血清（FBS）和1%青霉素-链霉素。HepG2细胞在Dulbecco改良的Eagle培养基（Corning, Inc.）中培养，补充有10%的FBS和1%的青霉素-链霉素，细胞在37℃和5%的二氧化碳中培养。
1. 野黄芩苷致敏浓度的筛选
采用CCK-8试剂盒（Dojindo，Kumamoto）评估HepG2细胞对野黄芩苷（上海源叶生物科技有限公司）细胞毒性的敏感性：在96孔板中培养的细胞用浓度为0-5μg/mL的野黄芩苷处理72小时。使用GraphPad Prism 9.0计算半数抑制浓度（IC50），选择IC50的10%作为野黄芩苷致敏浓度。不同野黄芩苷对2种HCC细胞的浓度对数-抑制率曲线如图1所示，经计算，针对HepG2细胞的增敏浓度为0.02μg/mL。
2. 野黄芩苷对125I抗增殖的影响
采用HepG2细胞系的增敏浓度的野黄芩苷，联合125I粒子体外照射模型共同处理细胞。将细胞分为：对照组、单SCU处理组、单125I处理组、SCU联合125I处理组。增敏浓度的SCU是由细胞的培养基配制而成，当把细胞的正常培养基更换为含有SCU的培养基后，随即将细胞放入125I粒子体外照射开始照射，照射时间为72h，初始活性水平为3.0 mCi，剂量率为3.412cGy/h。
在6孔板中培养HepG2细胞（2×105个/孔），分别用增敏浓度的SCU、125I粒子以及两者的联合处理细胞后，收集细胞并使用预冷的70%乙醇固定1小时，然后用PI和RNase A（BDBiosciences）染色，然后使用流式细胞仪（Beckman）检测。结果如图2所示：SCU和125I放射性粒子联合处理组的细胞周期停滞在G2/M期的程度大于单独使用125I放射性粒子或SCU的处理组。
3. 野黄芩苷对125I促进细胞凋亡的影响
在6孔板中培养HepG2细胞（2×105个/孔），分别用增敏浓度的SCU、125I粒子以及两者的联合处理细胞后收集。使用binding buffer重悬细胞，再加入5μL碘化丙啶（PI）和Annexin V-APC（Elabscience）或Annexin V-FITC（BD Biosciences）进行染色，避光20min后使用流式细胞仪（Beckman）检测。Annexin V-FITC/PI检测用于评估野黄芩苷对125I放射性粒子诱导的HCC细胞凋亡的影响，结果如图3所示：联合治疗组的细胞凋亡率显著高于野黄芩苷或125I放射性粒子单一治疗组。这说明，野黄芩苷促进了125I放射性粒子诱导的细胞凋亡。
4. 野黄芩苷对125I抑制细胞增殖的影响
在6孔板中培养HepG2细胞（2×105个/孔），分别用增敏浓度的SCU、125I粒子以及两者的联合处理细胞后收集，通过EdU实验检测细胞增殖。首先使用EdU溶液进行短时间孵育，随后使用细胞固定液室温孵育30分钟。然后使用Apollo染液和Hoechst33342染液分别对细胞进行染色。最后使用免疫荧光显微镜观察染色情况，结果如图4所示：联合治疗组的细胞增殖率显著低于野黄芩苷或125I放射性粒子单一治疗组。这说明，野黄芩苷促进了125I放射性粒子引发的细胞抗增殖作用。
Claims (2)
1.一种野黄芩苷作为125I放射治疗恶性肿瘤增敏剂的用途。
2.根据权利要求1所述的用途，其特征在于，所述恶性肿瘤选自胃癌、肠癌、肺癌或肝癌。
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| CN108392530A (zh) * | 2018-05-15 | 2018-08-14 | 中国药科大学 | 黄芩汤及其等效成分群用于增强结肠癌对伊立替康化疗敏感性的用途 |
| CN109394759A (zh) * | 2019-01-15 | 2019-03-01 | 王军 | 黄连素作为制备肿瘤放射治疗增敏药物中的应用 |
| CN113444765A (zh) * | 2020-03-25 | 2021-09-28 | 贵州医科大学 | 耐药性乳腺癌治疗药物及其筛选方法 |
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