摘要
目的探讨染料木素磺酸钠(genistein-3'-sodium sulfonate,GSS)对帕金森病(Parkinson disease,PD)模型小鼠运动功能及脑组织自噬水平的影响。方法将40只C57BL/6J小鼠按照随机数字表法分为对照组、模型组、GSS低剂量组(0.15 mg/kg)、GSS中剂量组(0.50 mg/kg)和GSS高剂量组(1.50 mg/kg),每组8只。模型组和GSS高、中、低剂量组小鼠采用腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶建立PD小鼠模型,建模后GSS高、中、低剂量组小鼠腹腔注射对应剂量的GSS(1次/d,共21 d),模型组小鼠则注射等体积0.9%氯化钠溶液(1次/d,共21 d),对照组小鼠正常饲养。21 d后通过步态分析实验、旷场实验、转棒实验、改良Y迷宫实验评估小鼠的运动和认知能力。Western blot法检测小鼠大脑皮质和纹状体组织LC3-Ⅱ和Beclin-1蛋白的表达水平。用SPSS 26.0软件对数据进行统计分析,正态分布多组间比较采用单因素方差分析,进一步两两比较采用LSD检验;非正态分布数据采用Kruskal-Wails H检验。结果(1)步态分析实验结果显示,5组小鼠左前肢、左后肢、右后肢的步长(F=5.93,6.21,3.78,均P<0.01)和正常步序比(H=14.409,P<0.01)均差异有统计学意义。模型组小鼠正常步序比低于对照组(P<0.05),GSS低、中、高剂量组正常步序比均高于模型组(均P<0.05)。(2)旷场实验中,5组小鼠的运动总路程和平均运动速度均差异有统计学意义(F=5.49,5.49,均P<0.01),模型组小鼠的运动总路程和平均运动速度均低于对照组(均P<0.05),GSS中、高剂量组小鼠的运动总路程[(2395.57±319.35)cm,(2386.51±396.00)cm]和平均运动速度[(7.98±1.06)cm/s,(7.95±1.32)cm/s]均高于模型组[(1863.31±278.96)cm,(6.21±0.93)cm/s]和GSS低剂量组[(1956.90±297.15)cm,(6.52±0.99)cm/s](均P<0.05)。(3)5组小鼠转棒实验中的掉落潜伏期和改良Y迷宫实验中的停留时间均差异有统计学意义(F=58.41,9.90,均P<0.01),模型组小鼠的掉落潜伏期和停留时间均低于对照组(均P<0.05),而GSS中、高剂量组的掉落潜伏期和停留时间均高于模型组和低剂量组(均P<0.05)。(4)Western blot结果显示,5组小鼠皮质和纹状体区LC3-Ⅱ/LC3-Ⅰ比值(F=8.17,15.47,均P<0.01)及Beclin-1蛋白(F=29.07,20.54,均P<0.01)的表达水平均差异有统计学意义。模型组小鼠的LC3-Ⅱ/LC3-Ⅰ比值及Beclin-1蛋白表达水平在皮质区[(0.51±0.14),(0.46±0.06)]和纹状体区[(0.58±0.09),(0.55±0.10)]均低于对照组[皮质区:(1.00±0.10),(1.00±0.05);纹状体区:(1.00±0.06),(1.00±0.25);均P<0.01];GSS中剂量组小鼠在皮质区和纹状体区的LC3-Ⅱ/LC3-Ⅰ比值均高于模型组和GSS低、高剂量组(均P<0.05);在皮质区模型组小鼠的Beclin-1蛋白表达水平低于GSS各剂量组(均P<0.05),在纹状体区模型组Beclin-1蛋白表达水平与GSS各剂量组比较,均差异无统计学意义(均P>0.05)。结论GSS能改善PD模型小鼠的运动和认知功能,其机制可能与上调小鼠大脑皮质和纹状体区的自噬活性有关。
ObjectiveTo explore the effects of genistein-3'-sodium sulfonate(GSS)on on motor function and brain autophagy levels in Parkinson disease(PD)model mice.MethodsForty C57BL/6J mice were randomly divided into control group,model group,low-dose GSS group(0.15 mg/kg),medium-dose GSS group(0.50 mg/kg)and high-dose GSS group(1.50 mg/kg),with 8 mice in each group.Mice in the model group and the high,medium,and low-dose GSS groups were injected intraperitoneally with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine to establish the PD mouse model,then mice in high,medium and low-close GSS group were intraperitoneally injected with corresponding doses of GSS(once a day for 21 days).The mice in model group were injected with equal volume 0.9% sodium chloride solution(once a day for 21 days),while the control group mice were fed normally.After 21 days,the motor and cognitive abilities of mice were evaluated by gait analysis,open field test,rotarod test,and modified Y maze test.Western blot was used to detect the expression levels of LC3-Ⅱand Beclin-1 proteins in the cerebral cortex and striatum tissues of mice.SPSS 26.0 software was used for data analysis.One-way ANOVA was used for normal distribution data comparison among multiple groups,and LSD test was used for further pairwise comparisons,while Kruskal-Wails H test was used for non normal distribution data comparison.Results(1)Gait analysis showed that there were statistically significant differences in the stride length of left forelimb,left hindlimb,right hindlimb(F=5.93,6.21,3.78,all P<0.01)and regularity index(H=14.409,P<0.01).The regularity index of the model group mice was lower than that of the control group(P<0.05),and the regularity indexes of the low,medium,and high-dose GSS groups were all higher than that of the model group(all P<0.05).(2)In the open field test,there were statistically significant differences in the total distance and speed of movement among the 5 groups(F=5.49,5.49,both P<0.01).The total distance and speed of movement in the model group were both lower than those in the control group(both P<0.05).The total distance and speed of movement in the medium-dose GSS group((2395.57±319.35)cm,(7.98±1.06)cm/s)and high-dose GSS group((2386.51±396.00)cm,(7.95±1.32)cm/s)were higher than those of the model group((1863.31±278.96)cm,(6.21±0.93)cm/s)and the low-dose GSS group((1956.90±297.15)cm,(6.52±0.99)cm/s)(all P<0.05).(3)In the rotarod test and modified Y maze test,there were significant differences in latency to fall and residence time among the 5 groups(F=58.41,9.90,both P<0.01).The latency to fall and residence time of model group were lower than those of control group(both P<0.05),while those in the medium-dose and high-dose GSS groups were higher than those in the model group and low-dose GSS group(all P<0.05).(4)Western blot results showed that there were significant differences in the expression levels of LC3-Ⅱ/LC3-Ⅰratio(F=8.17,15.47,both P<0.01)and Beclin-1 protein(F=29.07,20.54,both P<0.01)in cerebral cortex and striatum among the five groups.The LC3-Ⅱ/LC3-Ⅰratio and Beclin-1 protein levels in the cerebral cortex((0.51±0.14),(0.46±0.06))and striatum((0.58±0.09),(0.55±0.10))of the model group were lower than those in the control group(cerebral cortex:(1.00±0.10),(1.00±0.05),striatumm:(1.00±0.06),(1.00±0.25),all P<0.01).The LC3-Ⅱ/LC3-Ⅰratio and Beclin-1 protein in the medium-dose GSS group were higher than those in the model group,low-dose and high-dose GSS groups in both cerebral cortex and striatum(all P<0.05).The level of Beclin-1 of cerebral cortex in model group was lower than those in various doses of GSS group(all P<0.05).There were no statistically significant differences of Beclin-1 protein levels between the model group mice and various doses of GSS groups in striatum(all P>0.05).ConclusionGSS can improve the motor and cognitive functions of PD model mice,and the mechanism may be related to the upregulation of autophagic activity in the cerebral cortex and striatum of mice.
作者
袁博翔
顾乔乔
黄志华
薛进华
Yuan Boxiang;Gu Qiaoqiao;Huang Zhihua;Xue Jinhua(School of Basic Medical Sciences,Gannan Medical University,Ganzhou 341000,China)
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2024年第3期193-198,共6页
Chinese Journal of Behavioral Medicine and Brain Science
基金
江西省自然科学基金(20212BAB206001)。
关键词
帕金森病
认知功能
运动功能
染料木素磺酸钠
小鼠
Parkinson disease
Cognitive function
Motor function
Genistein-3'-sodium sulfonate
Mouse
作者简介
通信作者:薛进华,Email:xjhua@gmu.edu.cn。
