首页|石墨相氮化碳二维纳米片载洋川芎内酯I应用于缺血性脑卒中大鼠的治疗研究

石墨相氮化碳二维纳米片载洋川芎内酯I应用于缺血性脑卒中大鼠的治疗研究

扫码查看
目的:探讨石墨相氮化碳(g-C3N4)二维纳米片负载洋川芎内酯I(SEI)对缺血性脑卒中大鼠的治疗作用。方法:利用三聚氰胺通过热相反应生成层状g-C3N4,热剥酸剥后得到薄层g-C3N4纳米片并进行表征;将SEI负载到g-C3N4纳米片上,通过紫外可见吸收光谱测定上清中的SEI,计算g-C3N4负载SEI质量比,测定SEI-g-C3N4复合纳米片在1 h、3 h、6 h、12 h、24h和48h的药物释放情况;SD大鼠大脑中动脉闭塞(MCAO)造模前48h通过脑立体定位注射SEI-g-C3N4复合纳米片,造模后24h进行神经行为学评分,评分结束后取材分别进行TTC染色、TUNEL染色和HE染色。结果:通过表征实验,制备的g-C3N4纳米片尺寸小(约100nm)、比表面积大、分散性好、表面带正电荷(33。3 mV);紫外可见吸收光谱测定结果显示,g-C3N4已成功负载SEI,且SEI/g-C3N4质量比为15。06;药物释放结果显示,24 h时SEI的释放率接近50%;与sham组相比,MCAO组神经功能缺失评分显著升高(P<0。000 1),表明造模成功;与MCAO组相比,MCAO+SEI组神经功能缺失评分减少(P>0。05),MCAO+SEI-g-C3N4组神经功能缺失评分减少(P<0。001);与MCAO+SEI组相比,MCAO+SEI-g-C3N4组神经功能缺失评分减少(P<0。01);TTC染色结果表明,与sham组相比,MCAO组的脑梗死面积显著增加(P<0。000 1);与MCAO组相比,MCAO+SEI组梗死面积减少(P<0。000 1),MCAO+SEI-g-C3N4组脑梗死面积显著减少(P<0。000 1);与 MCAO+SEI 组相比,MCAO+SEI-g-C3N4 组脑梗死面积减少(P<0。001);TUNEL染色结果表明,与sham组相比,MCAO组的凋亡率升高(P<0。000 1),表明造模成功;与sham组相比,MCAO+SEI-g-C3N4组凋亡率升高(P>0。05);与MCA O组相比,MCAO+SEI-g-C3N4组凋亡率减少(P<0。001);体内生物安全性实验结果表明,与sham组相比,SEI组、SEI-g-C3N4组大鼠体重无统计学差异,且皮层脑组织结构和细胞形态无明显病理性变化。结论:成功制备了生物安全性较好的SEI-g-C3N4复合纳米片,且其能够减少MCAO大鼠的脑梗死面积,抑制细胞凋亡,减轻脑组织病理性变化,并且能改善大鼠的神经行为功能,表明SEI-g-C3N4复合纳米片可用于治疗缺血性脑卒中大鼠。
Study on the treatment of ischemic stroke rats with two-dimensional graphite carbon nitride nanosheets loaded with Senkyunolide I
Objective:To investigate the therapeutic effect of two-dimensional graphite phase carbon nitride(g-C3N4)nanosheets loaded with Senkyunolide I(SEI)on ischemic stroke rats.Methods:The layered g-C3N4 was formed by thermal reaction of melamine,and the thin layer g-C3N4 nanosheets were obtained and characterized af-ter thermal stripping and acid peeling.SEI was loaded onto g-C3N4 nanosheets,the SEI in the supernatant was de-termined by UV-vis absorption spectrum,the mass ratio of SEI loaded with g-C3N4 was calculated,and the drug release of SEI-g-C3N4 composite nanosheets was determined at 1 h,3 h,6h,12 h,24 h and 48 h.The SD rats with middle cerebral artery occlusion(MCAO)were injected with SEI-g-C3N4 composite nanosheets 48 h before the establishment of the model.Neurobehavioral score was performed 24 h after the establishment of the model.After the score,the samples were stained with TTC,TUNEL and HE,respectively.Results:Through the charac-terization experiment,the prepared g-C3N4 nanosheets had the advantages of small size(about 100 nm),large spe-cific surface area,good dispersion and positive surface charge(33.3 mV).The results of UV-vis absorption spec-tra showed that g-C3N4 had successfully loaded SEI and the mass ratio of SEI/g-C3N4 was 15.06.The drug release results showed that the release rate of SEI was close to 50%at 24 h.The neurological function score of MCAO group was significantly higher than that of sham group(P<0.0001),indicating that the modeling was successful.Compared with MCAO group,the neurological function score of MCAO+SEI group was decreased(P>0.05),and that of MCAO+SEI-g-C3N4 group was decreased(P<0.001).Compared with MCAO+SEI group,the neuro-logical function score of MCAO+SEI-g-C3N4 group was decreased(P<0.01).The results of TTC staining showed that compared with sham group,the cerebral infarction size of MCAO group was increased significantly(P<0.0001);compared with MCAO group,the cerebral infarction size of MCAO+SEI group was decreased(P<0.0001),and that of MCAO+SEI-g-C3N4 group was decreased significantly(P<0.0001).Compared with MCAO+SEI group,the cerebral infarction size of MCAO+SEI-g-C3N4 group was decreased(P<0.001).The results of TUNEL staining showed that the apoptosis rate of MCAO group was higher than that of sham group(P<0.0001),indicating that the modeling was successful,that of MCAO+SEI-g-C3N4 group was higher than that of sham group(P>0.05),and that of MCAO+SEI-g-C3N4 group was lower than that of MCAO group(P<0.001).The results of in vivo biosafety experiment showed that compared with sham group,there was no significant dif-ference in body weight between SEI group and SEI-g-C3N4 group,and no obvious pathological changes in corti-cal brain structure and cell morphology.Conclusion:The SEI-g-C3N4 composite nanosheets with good biosafety are successfully prepared,and they can reduce the cerebral infarction size,inhibit apoptosis,reduce the pathologi-cal changes of brain tissue,and improve the neurobehavioral function of MCAO rats,indicating that SEI-g-C3N4 composite nanosheets can be used in the treatment of ischemic stroke rats.

graphite carbon nitrideSenkyunolide Iischemic stroke rats

杨航、李金玲、刘斯佳

展开 >

广西医科大学基础医学院 广西脑科学研究重点实验室 广西高校生物分子医学研究重点实验室,南宁 530021

广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心 长寿与老年相关疾病教育部重点实验室 广西再生医学重点实验室,南宁 530021

石墨相氮化碳 洋川芎内酯I 缺血性脑卒中大鼠

国家自然科学基金资助项目广西重点研发计划资助项目

82360273桂科AB23026023

2024

广西医科大学学报
广西医科大学

广西医科大学学报

CSTPCD
影响因子:0.788
ISSN:1005-930X
年,卷(期):2024.41(6)