摘要
目的探讨低剂量氯胺酮对MPTP诱导帕金森病(PD)小鼠神经功能及神经炎症的影响。方法50只C57BL/6小鼠按照随机数字表法分为NC组、PD组、PD+Keta-L组、PD+Keta-M组和PD+Keta-H组,每组10只。PD+Keta-L、PD+Keta-M、PD+Keta-H组分别用25、50和75 mg/kg的氯胺酮腹腔注射处理,每天1次,共2 d,而后PD组及PD+Keta-L、PD+Keta-M、PD+Keta-H组腹腔注射MPTP 20 mg/kg,每天1次,连续20 d,建立亚急性PD小鼠模型。观察各组小鼠步态、改良神经功能缺损评分(MNSS);HE染色观察脑部神经细胞形态变化;免疫组化法检测神经细胞酪氨酸羟化酶(TH)表达,原位缺口末端标记(TUNEL)法检测神经细胞凋亡情况;酶联免疫吸附测定(ELISA)法检测脑组织肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-1β水平;Western blot法检测脑组织Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子(NF)-κB p65蛋白水平。结果与NC组相比,PD组步态异常,MNSS评分、脑组织神经细胞凋亡率以及TNF-α、IL-6、IL-1β水平升高,TH阳性细胞比例降低(P<0.05);与PD组相比,PD+Keta-L组步态改善,MNSS评分、神经细胞凋亡率、TNF-α、IL-6、IL-1β水平降低,TH阳性细胞比例升高(P<0.05),PD+Keta-M、PD+Keta-L组与PD组比较差异无统计学意义。PD组中TLR4、NF-κB p65和MyD88表达显著高于NC组,与PD组相比,PD+Keta-L组TLR4、NF-κB p65和MyD88均显著降低,PD+Keta-M组仅TLR4及MyD88降低(均P<0.05),而PD+Keta-H组无明显变化(P>0.05)。结论低剂量氯胺酮对PD小鼠具有一定神经保护作用。
Objective To investigate the effects of low-dose ketamine on the neurological function and neuroinflammation of MPTP induced Parkinson's disease(PD)mice.Methods Fifty C57BL/6 mice were divided into the NC group,PD group,PD+Keta-L group,PD+Keta-M group and PD+Keta-H group according to random number table method,with 10 mice in each group.PD+Keta-L,PD+Keta-M and PD+Keta-H groups were administered intraperitoneally with 25,50 and 75 mg/kg ketamine,respectively,once a day for 2 d.PD group,PD+Keta-L,PD+Keta-M and PD+Keta-H groups were intraperitoneally injected with MPTP 20 mg/kg once a day for 20 days to establish PD mouse models.The gait and modified Neurological Severity Scores(MNSS)were observed in each group.The brain tissues were collected,and HE staining was used to observe the morphological changes of brain nerve cells.The expression of tyrosine hydroxylase(TH)in nerve cells was detected by immunohistochemistry,and the apoptosis of nerve cells was detected by TUNEL assay.The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1βwere determined by ELISA.The protein levels of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88)and nuclear factor(NF)-κB p65 in brain tissues were determined by Western blot assay.Results Compared with NC group,the abnormal gaits,the MNSS score,the apoptosis of nerve cells and levels of TNF-α,IL-6 and IL-1βwere increased in PD group,and the proportion of TH positive cells was decreased(P<0.05).Compared with PD group,the abnormal gaits was improved,the MNSS score,nerve cell apoptosis rate,TNF-α,IL-6 and IL-1βlevels were decreased,,and the proportion of TH positive cells was increased in PD+Keta-L group(P<0.05).There were no significant differences in these indices between PD+Keta-M group,PD+Keta-L group and PD group(P>0.05).The expressions of TLR4,NF-κB p65 and MyD88 were significantly higher in the PD group than those in the NC group(P<0.05).Compared with the PD group,the expressions of TLR4,NF-κB p65 and MyD88 were significantly decreased in t
作者
宋俊杰
范军朝
陈英
陈勇
SONG Jun-jie;FAN Jun-chao;CHEN Ying;CHEN Yong(Department of Anesthesiology,the First Affiliated Hospital of Henan University,Kaifeng 475000,China)
出处
《天津医药》
CAS
北大核心
2021年第5期476-482,共7页
Tianjin Medical Journal
基金
河南省高等学校重点科研项目(16B320002,19A320021)。
关键词
帕金森病
氯胺酮
酪氨酸单氧化酶
细胞凋亡
神经保护
神经功能
神经炎症
Parkinson disease
ketamine
tyrosine 3-monooxygenase
apoptosis
neuroprotection
neurological function
neuroinflammation
