罗哌卡因诱导大鼠脊髓背角细胞凋亡中Fas/FasL的表达

Expression of Fas/FasL in Spinal Posterior Dorsal Horn Cell Apoptosis Induced by Ropivacaine in Rat

目的明确Fas/FasL在罗哌卡因诱导大鼠脊髓背角细胞凋亡中的表达情况,探讨罗哌卡因神经毒性的可能机制。方法成年雄性SD大鼠25只,随机分为空白对照组,模型对照组,罗哌卡因小、中、大剂量组,每组5只。除空白对照组外,其余各组大鼠均进行鞘内置管,并在造模3 d后分别鞘内注射0.9%氯化钠溶液或0.5%,1%,2%罗哌卡因(剂量均为0.12 mL·kg^(-1))。分别测量大鼠造模前1 d、给药前(造模后3 d)及鞘内给药后24 h后爪机械缩足反射(MWT)阈值;采用苏木精-伊红(HE)染色观察脊髓形态改变;TUNEL染色检测大鼠脊髓背角细胞凋亡水平;免疫组织化学染色检测各组脊髓组织中Fas、FasL的表达水平。结果与空白对照组比较,各模型组造模前后MWT无明显差异(P>0.05);给药后24 h,罗哌卡因中、大剂量组大鼠MWT较给药前明显升高(P<0.05)。HE染色结果显示,罗哌卡因小剂量组较模型对照组脊髓背角组织轻度水肿,罗哌卡因中、大剂量组间质明显水肿伴空泡增加。与模型对照组相比,TUNEL染色显示各罗哌卡因处理组脊髓背角凋亡细胞比例增大(P<0.05),罗哌卡因大剂量组较罗哌卡因小剂量组凋亡细胞比例增大更明显(P<0.05)。与模型对照组比较,罗哌卡因处理组Fas表达均升高(P<0.05);罗哌卡因大剂量组较罗哌卡因小剂量组,中剂量组升高更明显(P<0.05);而FasL的表达仅罗哌卡因大剂量组升高明显(P<0.05)。结论罗哌卡因可能通过上调Fas/FasL的表达诱导大鼠神经损伤和脊髓背角细胞凋亡。

Objective To investigate the expression of Fas/FasL in ropivacaine-induced apoptosis of rat spinal dorsal horn cells, and to explore the possible mechanism of neurotoxicity of ropivacaine. Methods Twenty-five adult male Sprague-Dawley rats were randomly divided into 5 groups: blank control group, model control group, low-,medium-and high-dose ropivacaine group, with 5 rats in each group. Except for the blank control group, the rats of other groups received intrathecal catheterization and 0.5%,1%,2% ropivacaine or 0.9% sodium chloride solution at the dosage of 0.12 mL·kg^(-1) were injected into the sheath after 3 days of model building.The morphological changes of spinal cord cells were detected by HE staining;TUNEL staining was used to detect the apoptosis of the spinal cord cells;Immunohistochemistry was used to detect the expression of Fas and FasL in the spinal cord tissue. Results Compared with the blank control group, there was no significant difference in MWT of the model control groups before and after intrathecal catheterization(P>0.05). And 24 hours after the injection, the MWT of rats in the medium-and high-dose ropivacaine groups was significantly higher than that before the intrathecal injection(P<0.05).HE staining showed that the low-dose ropivacaine group had little edema in the spinal dorsal horn tissue compared with the model control group had;and in the medium-and high-dose ropivacaine groups, the interstitial edema was observed and the vacuoles were increased(P<0.05).Compared with the model control group, the expression of Fas was increased in each ropivacaine treatment group(P<0.05),and the high-dose ropivacaine group was significantly higher than the low-and medium-dose groups(P<0.05);The expression of FasL was only significantly elevated in the high-dose ropivacaine group(P<0.05). Conclusion Ropivacaine may induce neuronal injury and apoptosis of spinal dorsal horn cells by up-regulating the expression of Fas/FasL.