【作者】 杨慧；

【导师】 吴祥根；

【作者基本信息】 青岛科技大学 ， 药学（专业学位）， 2022， 硕士

【摘要】 目的药物性肝损伤是临床上常见的急性肝损伤之一,引起药物性肝损伤的最常见的原因是过量使用对乙酰氨基酚。目前,临床上对于对乙酰氨基酚引起的急性肝损伤只有N-乙酰半胱氨酸一种治疗药物,但是N-乙酰半胱氨酸对于晚期的患者作用较弱。因此,研究新的治疗药物具有很重要的意义。本研究将构建甘草酸二钾-漆黄素前体胶束这一创新制剂,并使用对乙酰氨基酚构建小鼠急性肝损伤模型,评价其对于急性肝损伤的治疗效果。方法1.使用溶剂蒸发法构建甘草酸二钾-漆黄素前体胶束,优化处方,并对其固体及溶液特性进行表征;评价其储存稳定性;采用FRAP法测定其抗氧化能力;使用溶血实验考察甘草酸二钾前体胶束的安全性。2.测定甘草酸二钾-漆黄素前体胶束在不同介质中的溶解度,并通过透析袋实验对其体外释放特性进行考察;采取体外平行人工膜渗透实验对前体胶束渗透性进行考察;使用香豆素6作为模型药物通过荧光强度考察其体内吸收特性。3.使用对乙酰氨基酚构建小鼠肝损伤模型。观察小鼠72小时内的存活情况;收集小鼠肝脏、脾脏并称重,观察肝脏外观,计算肝指数以及脾指数;对小鼠血清中ALT和AST进行测定;石蜡切片H&E染色考察肝脏病理变化;TUNEL法对小鼠肝脏细胞凋亡情况进行评价;小鼠肝脏中氧化应激和炎症因子水平进行考察。结果1.成功制备甘草酸二钾-漆黄素前体胶束。前体胶束水溶性良好,溶液呈黄色透明状,粒径为84.17±3.88 nm,包封率达到99.65±0.78%;储存稳定性良好;抗氧化能力较强;无溶血性,安全性良好。2.在水中,甘草酸二钾-漆黄素前体胶束中漆黄素的溶解度是常规漆黄素的1804倍;体外释放结果表明,在5小时前体胶束的累积释放量达到了92.87±2.79%;在3.5小时前体胶束中漆黄素的渗透量为267.30±14.53μg,是漆黄素混悬液的21.2倍;体内吸收实验表明,甘草酸二钾的包封显著提高了模型药物在肠道的吸收。3.使用对乙酰氨基酚成功建立小鼠肝损伤模型。甘草酸二钾-漆黄素前体胶束组72 h存活率为83.3%;肝脏外观与健康对照组相似;血清中ALT和AST恢复到正常水平(与健康对照组相比P>0.05);与模型组相比,肝脏中的氧化应激和炎症因子水平均显著降低(P<0.05)。结论甘草酸二钾-漆黄素前体胶束显著提高了药物的溶解度和体内吸收,对于对乙酰氨基酚引起的小鼠急性肝损伤具有保护作用,具有良好的进一步研究前景。更多还原

【Abstract】 ObjectiveDrug-induced liver injury is one of the most common acute liver injuries in clinical practice,and the most common cause of drug-induced liver injury is the overdose of acetaminophen.At present,N-acetylcysteine is the only clinical treatment for acute liver injury caused by acetaminophen,but N-acetylcysteine has a weak effect on advanced patients.Therefore,it has great significance to develop new therapeutic drug.In this study,a novel dipotassium glycyrrhizinate-fisetin premicelles(DG-FIT)was fabricated,acetaminophen was used to establish a mouse model of acute liver injury,and this animal model was used to evaluate the therapeutic effect of DG-FIT.Methods1.Solvent evaporation method was used to prepare DG-FIT,and the formulation of DG-FIT was optimized,and characterize its solids and solutions;the storage stability was evaluated;the antioxidant activity of DG-FIT was determined by FRAP method;the safety of DG-FIT was evaluated by hemolysis test.2.The solubility of DG-FIT in different media was determined,and the in vitro release characteristics were investigated by dialysis bag experiment;the permeability of DG-FIT was investigated by in vitro parallel artificial membrane permeation experiments;as a model drug,coumarin-6 was used to investigate its in vivo absorption characteristics by fluorescence intensity.3.A mouse model of liver injury was established using acetaminophen.The survival of the mice within 72 hours was observed;the liver and spleen of the mice were collected and weighed,the appearance of the liver was observed,the liver index and the spleen index were calculated;the serum ALT and AST of the mice were determined;the H&E staining of paraffin sections was used to investigate the pathological changes of the liver;TUNEL method was used to evaluate the apoptosis of liver cells in mice;the levels of oxidative stress and inflammatory factors in the liver of mice were investigated.Results1.DG-FIT was successfully prepared.DG-FIT could be easily dissolved in water to form a clear and yellow micelle solution;the particle size of DG-FIT was84.17±3.88 nm,and the encapsulation efficiency reached 99.65±0.78 %;it had good storage stability and strong antioxidant capacity;this premicelles showed well safety with no hemolysis occurred.2.DG-FIT significantly increased the solubility of fisetin in water.In vitro release experiments,the accumulative release rate of premicelles reached92.87±2.79 % at 5 h;in vitro absorption experiments,the permeation amount of fisetin in the premicelles reached 267.30±14.53 μg at 3.5 h,which was 21.2 times that of the suspension group;in vivo absorption experiment showed that dipotassium glycyrrhizinate encapsulation significantly improved the intestinal absorption of model drug.3.A mouse liver injury model was successfully established by acetaminophen.The 72-hour survival rate of the DG-FIT group was 83.3 %,the appearance of the liver was similar to that of the healthy control group;the serum ALT and AST returned to normal levels(P>0.05 compared with the healthy control group);compared with the control group,the levels of oxidative stress and inflammatory factors in the liver were significantly decreased(P<0.05).ConclusionDG-FIT can significantly improved the solubility and in vivo absorption of the drug,and has a good development prospect for the protective effect of acetaminophen-induced acute liver injury in mice.更多还原