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目的对抗癌药物索拉非尼体外抗多房棘球蚴的效果进行评价。方法从感染小鼠体内收集多房棘球蚴小囊泡,加入预先接种大鼠肝细胞的DMEM培养液(含10%胎牛血清、100 U/m L青霉素、100 mg/L链霉素)中,37℃、5% CO_2培养2~3个月后,选取直径2~4 mm的完整包囊用于药物效果评价实验。实验分阿苯达唑组(阳性对照),阿苯达唑终浓度分别为10μmol/L和30μmol/L;索拉非尼组(实验组),索拉非尼终浓度分别为10μmol/L和30μmol/L;DMSO组(溶剂对照组),加DMSO(1︰9溶于PBS中);PBS组(阴性对照),仅加PBS。每孔取25~30个包囊,每个浓度设3个重复。药物处理48、72、96、120、144和168 h后,光学显微镜和扫描电镜观察包囊形态结构变化,计算包囊抑制率,对其体外抗棘球蚴效果进行评价。结果10、30μmol/L索拉非尼处理48、72、96、120、144和168 h后,包囊的抑制率分别为6.6%、42.4%、68.5%、77.4%、84.0%、89.5%和7.1%、45.6%、70.9%、82.6%、84.0%、89.9%。10、30μmol/L阿苯达唑在每个时间点的包囊抑制率分别为2.6%、6.6%、21.4%、47.8%、59.9%、70.6%和3.8%、12.7%、27.0%、51.4%、54.0%、73.0%。DMSO和PBS对包囊没有明显的抑制效果。索拉非尼的抑制效果均高于阿苯达唑,从处理后72 h起,索拉非尼和阿苯达唑的抑制率差异有统计学意义(P<0.05)。光学显微镜观察表明,索拉非尼处理后,包囊向内部陷塌,形成“日全食”状;扫描电镜观察发现,索拉非尼处理后,包囊内部生发层细胞脱落,完全失去活性。结论索拉非尼显示出较高的包囊抑制效果,是棘球蚴病治疗的潜在药物。
Objective To evaluate the efficacy of sorafenib against Echinococcus multilocularis in vitro. Methods Echinococcus multilocularis vesicles were collected from infected mice and cultured in DMEM containing 10% fetal bovine serum, 100 U / mL penicillin and 100 mg / L streptomycin, After complete incubation for 2 to 3 months at 37 ° C and 5% CO 2, an intact capsule with a diameter of 2 to 4 mm was selected for drug effect evaluation. The experimental albendazole group (positive control), albendazole final concentrations were 10μmol / L and 30μmol / L; the sorafenib group (experimental group), sorafenib final concentrations were 10μmol / L and 30μmol / L DMSO group (solvent control group), plus DMSO (1︰9 in PBS); PBS group (negative control), only PBS. Take 25 to 30 cysts per well and set up 3 replicates for each concentration. 48, 72, 96, 120, 144 and 168 h after drug treatment, the morphological changes of the cysts were observed under light microscope and scanning electron microscopy. The inhibitory rate of cysts was calculated and the anti-hydatid cyst in vitro was evaluated. Results The inhibitory rates of encapsulation after 10, 30μmol / L sorafenib treatment for 48, 72, 96, 120, 144 and 168 h were 6.6%, 42.4%, 68.5%, 77.4%, 84.0%, 89.5% and 7.1%, 45.6%, 70.9%, 82.6%, 84.0%, 89.9%. The encapsulation inhibition rates of albendazole 10 and 30μmol / L at each time point were 2.6%, 6.6%, 21.4%, 47.8%, 59.9%, 70.6% and 3.8%, 12.7%, 27.0% and 51.4% , 54.0%, 73.0%. DMSO and PBS showed no significant inhibitory effect on the cysts. The inhibitory effect of sorafenib was higher than that of albendazole. The inhibition rates of sorafenib and albendazole were statistically significant at 72 h after treatment (P <0.05). Optical microscopy showed that after sorafenib treatment, the capsule collapsed to the interior to form a “total solar eclipse” shape. Scanning electron microscopy showed that after sorafenib treatment, the cells in the inner germinal layer of the cyst shed and disappeared completely active. Conclusion Sorafenib shows a high cyst inhibitory effect and is a potential drug for the treatment of hydatid disease.