Jia-Qi Chen, Feng L I, Peng-Kun Xue, Yuan-Yuan L I, Sha-Sha Wang, Hua-Hua Wang, Chun-Yue H U, Xia-Li Zhu
School of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China.
School of Basic Medicine, Henan University of Chinese Medicine Zhengzhou 450046, China.
Zhongguo Zhong Yao Za Zhi. 2020 Oct;45(19):4617-4624. doi: 10.19540/j.cnki.cjcmm.20200706.306.
With matrine(MAT) as the model drug, to prepare nano graphene oxide(NGO)-based MAT in situ gel(MAT-NGO-gel), a kind of drug for tumor treatment in combination with phototheraphy, and investigate the physicochemical properties and anti-tumor effects in vivo of MAT-NGO-gel. First, HPLC method was established to measure the content of MAT in the gel. The ultrasonic method was used to load MAT onto the surface of NGO, and then poloxamer 188 and poloxamer 407 were chosen as the main materials to prepare MAT-NGO-gel. The optimum prescription was selected with the gelation temperature as the index. Finally, the drug loading rate, micromorphology, phototherrmal conversion characteristics and drug release in vitro of MAT-NGO-gel were characterized. In the optimized prescription, the concentration of poloxamer 188 and poloxamer 407 was 2% and 20% respectively, and the mass ratio of NGO and MAT was 1∶1. The gelation temperature and drug loading rate of MAT-NGO-gel prepared by the optimal prescription process was 37.5 ℃ and 16.7%. Under 808 nm laser irradiation, MAT-NGO-gel showed obvious concentration-and time-dependent photothermal conversion characteristics. In vitro release experiments showed that MAT-NGO-gel had temperature-dependent release characteristics. The pharmacodynamics of MAT solution, NGO-gel and MAT-NGO-gel were studied by using S180 tumor-bearing mice and 808 nm laser. The relative tumor volume and body weight of the tumor-bearing mice were plotted over time. After the experiment, the tumor tissues of each group were taken and the histopathological changes were observed by HE staining. The results of pharmacodynamic studies demonstrated that when compared with NS group and NGO-gel group, the body weights of mice in MAT-NGO-gel group and MAT-NGO-gel + laser group were higher, and the relative tumor volume growth was slower. The results of HE stained pathological sections showed that the tumor cells count for the mice in MAT-NGO-gel group and MAT-NGO-gel + laser group was significantly reduced, with obvious nuclear fragmentation and nucleolysis in these two groups. These results suggested that MAT-NGO-gel, especially combined with 808 nm laser, had stronger anti-tumor activity in vivo. The prescription process of MAT-NGO-gel in this experiment was stable and feasible. As compared with MAT solution, MAT-NGO-gel showed obvious sustained and temperature-dependent drug release characteristics. MAT-NGO-gel had much more obvious anti-tumor activity in vivo when combined with 808 nm laser irradiation. This study could provide certain theoretical basis for the therapy of malignant tumor with multiple mechanisms.
以苦参碱（MAT）为模型药物，制备用于肿瘤治疗且兼具光热疗法的纳米氧化石墨烯（NGO）基MAT原位凝胶（MAT-NGO-凝胶），并研究MAT-NGO-凝胶的理化性质及体内抗肿瘤效果。首先，建立高效液相色谱（HPLC）法测定凝胶中MAT的含量。采用超声法将MAT负载于NGO表面，然后选用泊洛沙姆188和泊洛沙姆407作为主要材料制备MAT-NGO-凝胶。以凝胶化温度为指标筛选最佳处方。最后，对MAT-NGO-凝胶的载药率、微观形态、光热转换特性及体外药物释放进行表征。在优化处方中，泊洛沙姆188和泊洛沙姆407的浓度分别为2%和20%，NGO与MAT的质量比为1∶1。通过最佳处方工艺制备的MAT-NGO-凝胶的凝胶化温度和载药率分别为37.5℃和16.7%。在808nm激光照射下，MAT-NGO-凝胶呈现出明显的浓度和时间依赖性光热转换特性。体外释放实验表明，MAT-NGO-凝胶具有温度依赖性释放特性。采用S180荷瘤小鼠和808nm激光研究MAT溶液、NGO-凝胶和MAT-NGO-凝胶的药效学。绘制荷瘤小鼠的相对肿瘤体积和体重随时间的变化曲线。实验结束后，取各组肿瘤组织，通过苏木精-伊红（HE）染色观察组织病理学变化。药效学研究结果表明，与生理盐水组和NGO-凝胶组相比，MAT-NGO-凝胶组和MAT-NGO-凝胶+激光组小鼠的体重更高，相对肿瘤体积增长更慢。HE染色病理切片结果显示，MAT-NGO-凝胶组和MAT-NGO-凝胶+激光组小鼠的肿瘤细胞数量显著减少，这两组均有明显的核碎裂和核溶解现象。这些结果表明，MAT-NGO-凝胶，尤其是与808nm激光联合使用时，在体内具有更强的抗肿瘤活性。本实验中MAT-NGO-凝胶的处方工艺稳定可行。与MAT溶液相比，MAT-NGO-凝胶表现出明显的缓释和温度依赖性药物释放特性。MAT-NGO-凝胶与808nm激光照射联合使用时在体内具有更明显的抗肿瘤活性。本研究可为多机制治疗恶性肿瘤提供一定的理论依据。
