脑对流增强给药对老年大鼠脑细胞外间隙微观结构的影响

Effect of convection enhanced delivery on the microstructure of brain extracellular space in aged rats

目的:对比研究经细胞外间隙(extracellular space,ECS)途径的脑对流增强给药(convection enhanced delivery,CED)在进行脑病微创治疗时,不同给药速率下成年大鼠与老年大鼠脑ECS结构参数及局部药物分布的改变。方法:36只SD雄性大鼠按照月龄分为成年大鼠组(2~8月龄,18只)和老年大鼠组(18~24月龄,18只),每组再按照不同的给药速率(0.1μL/min、0.2μL/min、0.3μL/min)随机分为3个亚组,每亚组6只。采用立体定位注射法分别在各组鼠脑尾状核区导入浓度为10 mmol/L的磁示踪剂钆-二乙三胺五乙酸(gadolinium-diethylene triamine pentaacetic acid,Gd-DTPA)后,应用磁示踪法动态采集Gd-DTPA在脑间质系统(brain interstitial system,ISS)中的扩散和分布图像。利用自主研发的MRI影像测量分析系统软件对所获得的图像进行处理和分析,可获得各组大鼠脑尾状核区ECS内的有效扩散系数(DECS)、清除率、容积占比和半衰期(T1/2)等参数。比较分析在不同给药速率下,老年大鼠与成年大鼠脑ECS内药物清除以及ECS结构功能的影响和差异。应用磁示踪DECS-mapping技术观察示踪剂在尾状核区的分布引流情况。结果:0.1μL/min注药速率下,与成年大鼠相比,老年大鼠的容积占比增加(18.20%±0.04%vs.17.20%±0.03%,t=3.752,P=0.004),迂曲度下降(1.63±0.04 vs.1.78±0.09,t=-3.680,P=0.004),药物清除速率下降[(1.94±0.68)mm2/s vs.(3.25±0.43)mm2/s,t=-3.971,P=0.003],ECS内分子扩散速率增快[(3.99±0.21)×10-4mm2/s vs.(3.36±0.37)×10-4mm2/s,t=3.663,P=0.004]。注药速率增加到0.2μL/min时,老年大鼠ECS内药物清除减慢[(2.53±0.45)mmol/L vs.(3.37±0.72)mmol/L,t=-1.828,P=0.021],但容积占比、ECS内分子扩散和宏观药物代谢参数无明显差异。注药速率增加到0.3μL/min时,老年大鼠容积占比减小(17.20%±0.03%vs.18.20%±0.05%,t=-0.869,P=0.045),ECS内药物清除明显加快[(4.04±0.76)mmol/L vs.(3.26±0.55)mmol/L,t=1.786,P=0.014],迂曲度和ECS内分子扩散速率无明显差异。结论:老年脑CED给药在不同速率时ECS内药物清除及ECS结构参数发生改变,0.2μL/min速率下CED给药对老年脑ECS影响最小。应用CED进行脑病治疗时应综合考虑年龄和注药速率等因素的影响,经ECS途径给药进行脑病微创治疗时应制定个体化临床治疗方案。

Objective:To compare the changes of extracellular space(ECS)structure and local drug distribution in adult brain and aged brain at different drug delivery rates in minimally invasive treatment of encephalopathy by convection enhanced delivery(CED)via ECS pathway.Methods:Thirty-six SD male rats were divided into adult rats group(2-8 months,n=18)and aged rats group(18-24 months,n=18)according to the age of the month.According to the drug rates(0.1μL/min,0.2μL/min,and 0.3μL/min),they were randomly divided into 3 subgroups,6 in each subgroup.Gadolinium-diethylene triamine pentaacetic acid(Gd-DTPA)with a concentration of 10 mmol/L were introduced into the caudate nucleus of each group of rats by stereotactic injection.Tracer-based magnetic resonance imaging(MRI)was used to dynamically monitor the diffusion and distribution images of the Gd-DTPA in the brain interstitial system(ISS).Using the self-developed MRI image measurement and analysis system software to process and analyze the obtained images,the diffusion coefficient,clearance rate,volume fraction,and half-life of each group of rats in the caudate nucleus ECS could be acquired.The effects and differences of drug clearance and ECS structural function in the brain of aged rats and adult rats were compared and analyzed at different drug delivery rates.Magnetic tracer DECS-mapping technique was used to observe the distribution and drainage of tracer in caudate nucleus.Results:At the injection rate of 0.1μL/min,the volume fraction in the aged rats was increased compared with that in the adult rats(18.20%±0.04%vs.17.20%±0.03%,t=3.752,P=0.004),and the degree of tortuosity was decreased(1.63±0.04 vs.1.78±0.09,t=-3.680,P=0.004),the drug clearance rate was decreased[(1.94±0.68)mm2/s vs.(3.25±0.43)mm2/s,t=-3.971,P=0.003],and the molecular diffusion in ECS was increased[(3.99±0.21)×10-4 mm2/s vs.(3.36±0.37)×10-4 mm2/s,t=3.663,P=0.004].When the rate of injection increased to 0.2μL/min,the drug clearance in ECS of the aged rats was slowed down[(2.53±0.45)mmol/L vs.(3.37±0.72)mmol/L,t=-1.828,P=0.021].However,there were no significant differences in volume fraction,molecular diffusion in ECS and macroscopic drug metabolism parameters.When the rate of injection increased to 0.3μL/min,the volume fraction in the aged rats was decreased(17.20%±0.03%vs.18.20%±0.05%,t=-0.869,P=0.045),and the drug clearance rate in ECS was significantly accelerated[(4.04±0.76)mmol/L vs.(3.26±0.55)mmol/L,t=1.786,P=0.014],and there was no significant difference in tortuosity and the rate of molecular diffusion in the ECS.Conclusion:The drug clearance and ECS structural parameters of brain ECS in aged brain with CED administration were changed at different rates,and it has the least effect on ECS in the aged brain at the injection rate of 0.2μL/min.For the application of CED for the treatment of encephalopathy,we should consider the influence of factors such as age and injection rate,and provide reference for the development of individualized clinical treatment plan for minimally invasive treatment of encephalopathy via ECS pathway.