积雪草酸调节cAMP/PKA信号通路对罗哌卡因诱导新生大鼠神经毒性的影响  

Influence of asiatic acid on ropivacaine induced neurotoxicity in neonatal rats by regulating cAMP/PKA signaling pathway

作　　者：何洁[1] 李秀泽[1] 文竹[1] HE Jie;LI Xiuze;WEN Zhu(Department of Anesthesia,Mianyang Central Hospital,Mianyang,Sichuan 621000,China)

机构地区：[1]绵阳市中心医院麻醉科,四川绵阳621000

出　　处：《中国优生与遗传杂志》2023年第8期1575-1580,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的 探究积雪草酸(AA)调节环磷酸腺苷(cAMP)/蛋白激酶A(PKA)信号通路对罗哌卡因诱导新生大鼠神经毒性的影响。方法 将SPF级SD新生大鼠随机分为假手术组(Sham组)、模型组(Model组)、低剂量AA组(AA-L组,50 mg/kg)、高剂量AA组(AA-H组,100 mg/kg)和高剂量AA+PKA抑制剂H-89组(AA-H+H-89组,100 mg/kg AA+2 mg/kg H-89),每组12只。采用鞘内注射罗哌卡因构建新生大鼠脊髓神经毒性模型。采用BBB评分、斜板实验评价各组大鼠运动功能。H-E染色观察各组大鼠脊髓组织病理学变化。酶联免疫吸附(ELISA)法检测各组大鼠脊髓组织cAMP水平。TUNEL染色观察各组大鼠脊髓组织细胞凋亡情况。实时荧光定量PCR(RT-qPCR)法检测各组大鼠脊髓组织中PKA mRNA的表达。Western blot检测各组大鼠脊髓组织PKA、Bax、Caspase-3、cAMP应答元件结合蛋白(CREB)、p-CREB、脑源性神经营养因子(BDNF)蛋白表达。结果 与Sham组相比,Model组大鼠BBB评分、斜板角度、脊髓组织cAMP含量、PKAmRNA和蛋白表达、p-CREB、BDNF蛋白表达显著下降(P<0.05),脊髓组织细胞凋亡率、Bax、Caspase蛋白表达显著上升(P<0.05),脊髓组织结构出现损伤。与Model组相比,AA-H组大鼠BBB评分、斜板角度、脊髓组织cAMP含量、PKA mRNA和蛋白表达、p-CREB、BDNF蛋白表达显著升高(P<0.05),脊髓组织细胞凋亡率、Bax、Caspase蛋白表达显著下降(P<0.05),脊髓组织损伤减轻。H-89减轻了AA对罗哌卡因诱导的神经毒性的改善。结论 AA可能通过激活cAMP/PKA信号通路减轻罗哌卡因诱导的新生大鼠神经毒性。Objective To investigate the influence of asiatic acid(AA)on the neurotoxicity induced by ropivacaine in neonatal rats by regulating cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)signaling pathway.Methods SPF SD newborn rats were randomly divided into sham operation group(Sham group),model group(Model group),low-dose AA group(AA-L group,50 mg/kg),high-dose AA group(AA-H group,100 mg/kg),and high-dose AA+PKA inhibitor H-89 group(AA-H+H-89 group,100 mg/kg AA+2 mg/kg H-89),with 12 rats in each group.The spinal cord neurotoxicity model of neo-natal rats was established by intrathecal injection of ropivacaine.BBB score and inclined plate test were used to evaluate the motor function of rats in each group.Histopathological changes of spinal cord were observed by HE staining.Enzyme linked immunosorbent assay(ELISA)was used to detect the cAMP level in spinal cord tissue of rats in each group.TUNEL staining was used to observe the apoptosis of spinal cord tissue cells of rats in each group.The expression of PKA mRNA in spinal cord tissue of rats in each group was detected by real-time fluorescent quantitative PCR(RT-qPCR).The expression of PKA,Bax,Caspase-3,cAMP response element binding protein(CREB),p-CREB and brain-derived neurotrophic factor(BDNF)in spinal cord tissue of rats in each group was detected by Western blot.Results Compared with Sham group,the BBB score,oblique plate angle,cAMP content in spinal cord,PKA mRNA and protein expression,p-CREB,BDNF protein expression in model group decreased obviously(P<0.05),the apoptosis rate,Bax,Caspase protein expression in spinal cord tissue increased obvi-ously(P<0.05),the spinal cord tissue structure was damaged.Compared with Model group,the BBB score,oblique plate angle,cAMP content in spinal cord,PKA mRNA and protein expression,p-CREB,BDNF protein expression in AA-H group obvi-ously increased(P<0.05),the apoptosis rate,Bax,Caspase protein expression of spinal cord tissue decreased obviously(P<0.05),the injury of spinal cord tissue was alleviated.H-89 alleviat

关 键 词：积雪草酸 环磷酸腺苷/蛋白激酶A信号通路 罗哌卡因 神经毒性 

分 类 号：R285.5[医药卫生—中药学]