经皮椎间孔注射多柔比星与多柔比星脂质体对大鼠后肢行为学影响的比较

Comparison of the effect of percutaneous intervertebral foramen injection of doxorubicin and pegylated liposome doxorubicin on hind limb behavior in ratsLI Limei1,2, CHEN Jun3, ZHENG Baosen4, ZHANG Zhili2, SUN Wei3, YANG Fan3, YU Wenli5 △

摘要:

目的 比较经皮椎间孔注射多柔比星与多柔比星脂质体对成年大鼠后肢行为学的影响，探讨多柔比星脂质体对背根神经节的镇痛作用。 方法 健康雄性SD大鼠52只，体重200-250 g，3月龄，采用随机数字表法，将大鼠随机分为4组(n=13)：生理盐水组（Vehicle组）、隐形脂质体组（Liposome组）、0.2%多柔比星组（DOX组）及0.2%多柔比星脂质体组（PLD组）。各组均采用经皮椎间孔注射方法，于大鼠左侧L4/5、L5/6椎间孔分别注射生理盐水、隐形脂质体注射液、多柔比星及多柔比星脂质体10ul，术后3天，DOX组和PLD组各随机抽取1只大鼠行4%多聚甲醛灌流取材、冰冻切片后，共聚焦荧光显微镜观察多柔比星自发荧光情况；于给药前及给药后1天、3天、5天、7天、14天、21天、28天测定大鼠机械缩足反应阈值（PWMT）和热缩足潜伏期（PWTL），并观察给药后大鼠后肢的运动情况。 结果 荧光显微镜显示，DOX组和PLD组注药侧DRG内可见大量红色多柔比星自发荧光分布，非注药侧DRG内无多柔比星自发荧光。与Vehicle组相比，Liposome组给药后各时点PWMT值和PWTL值无明显差异（P>0.05）；与Vehicle组和Liposome组相比，DOX组 PWMT值给药后1周内、PLD组给药后2周内下降，与基础值比较，差异无统计学意义（P>0.05）；之后呈逐渐上升趋势，至给药后28天增高最明显（P<0.05）；DOX组和PLD组给药后1天PWTL值下降，与基础值比较差异有统计学意义（P<0.05），余各时点组内及组间比较差异均无统计学意义（P>0.05）。注药后28天，DOX组轻微跛行率为30%，其余各组大鼠后肢运动功能均正常（P>0.05）。 结论 经皮椎间孔注射多柔比星脂质体对背根神经节能够产生和多柔比星相似的镇痛作用，且不影响下肢运动功能。

Abstract:

Objective To compare the effects of percutaneous intervertebral foramen injection of doxorubicin and pegylated liposomes doxorubicin on the behavior of hind limbs in adult rats, and to explore the analgesic effect of doxorubicin liposomes on dorsal root ganglion. Methods 52 healthy male SD rats, weighing 200-250 g, aged 3 months, were randomly divided into 4 groups (n=13): normal saline group (Vehicle Group), stealth liposome group (Liposome group), 0.2% doxorubicin group (DOX group) and 0.2% pegylated liposomes doxorubicin group (PLD group). According to the different groups, normal saline, stealth liposome, doxorubicin and pegylated liposomes doxorubicin 10ul were injected respectively into the intervertebral foramen of the left L4/5 and L5/6 of rats by percutaneous intervertebral foramen injection. Three days after administration, 1 rat in DOX group and PLD group was randomly selected for 4% polymethylaldehyde perfusion and frozen section. The spontaneous fluorescence of doxorubicin was observed by fluorescence microscope. The paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) of rats were measured before and after administration at 1, 3, 5, 7, 14, 21 and 28 days, and the movement of hind limbs of rats after administration was also observed. Results Fluorescence microscope showed that a large number of orange doxorubicin autofluorescence distribution could be seen in the DRG on the injection side of DOX group and PLD group, no autofluorescence distribution could be seen on the contralateral DRG. Compared with vehicle group, there was no significant difference in PWMT and PWTL values at each time point after administration in liposome group (P>0.05); Compared with vehicle group and liposome group, PWMT value in DOX group decreased within 1 week after administration, while that within 2 weeks in PLD group after administration, and there were no significant difference compared with the basic value (P >0.05); then, the PWMT showed a gradual upward trend, and the increase was most obvious at 28 days after administration (P <0.05); PWTL value of DOX group and PLD group decreased one day after administration, and the difference was statistically significant compared with the basic value (P <0.05), and there was no statistically significant difference between the other time points (P >0.05). The hind limb movement of all rats was normal after administration. Conclusion Percutaneous injection of pegylated liposomes doxorubicin into intervertebral foramen can produce similar analgesia effect to doxorubicin, and does not affect the motor function of hind limbs.