中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2010年
2期
204-208
,共5页
曹碧茵%孔岩%徐侣%杨亚萍%刘春风
曹碧茵%孔巖%徐侶%楊亞萍%劉春風
조벽인%공암%서려%양아평%류춘풍
芍药苷%多巴胺能神经元%黑质脑片%MPP~+%α-synuclein%大鼠
芍藥苷%多巴胺能神經元%黑質腦片%MPP~+%α-synuclein%大鼠
작약감%다파알능신경원%흑질뇌편%MPP~+%α-synuclein%대서
paeoniflorin%dopaminergic neuron%brain slice of substantia nigravia%MPP~+%α-synuclein%rat
目的 研究芍药苷(paeoniflorin,PF)对MPP~+所致大鼠黑质脑片多巴胺能神经元损伤的保护作用,并观察PF对α-synuclein(α-syn) mRNA表达水平的影响.方法 离体培养SD大鼠黑质器官型脑片,培养的d 10给予不同浓度的MPP~+(0.1、0.5、1.0 mmol·L~(-1))处理24 h,并在0.5 mmol·L~(-1) MPP~+处理的基础上,给予1 μmol·L~(-1)或10 μmol·L~(-1)的PF进行干预.免疫组化法计数黑质致密部酪氨酸羟化酶(tyrosine hydroxylase,TH)阳性细胞数,定量RT-PCR法检测α-syn mRNA的表达丰度.结果 MPP~+(0.1、0.5、1.0 mmol·L~(-1))处理导致黑质致密部TH+细胞数较正常对照组减少,差异均有统计学意义(P<0.05或P<0.01);MPP~+(0.5 mmol·L~(-1))使α-syn mRNA的表达水平明显升高(P<0.01).PF(10 μmol·L~(-1))有效增加了TH+细胞的数量(P<0.01),并明显下调了α-syn mRNA的表达(P<0.05).结论 PF能有效拮抗MPP~+导致的多巴胺能神经元受损死亡,其机制可能与下调α-syn的表达有关.
目的 研究芍藥苷(paeoniflorin,PF)對MPP~+所緻大鼠黑質腦片多巴胺能神經元損傷的保護作用,併觀察PF對α-synuclein(α-syn) mRNA錶達水平的影響.方法 離體培養SD大鼠黑質器官型腦片,培養的d 10給予不同濃度的MPP~+(0.1、0.5、1.0 mmol·L~(-1))處理24 h,併在0.5 mmol·L~(-1) MPP~+處理的基礎上,給予1 μmol·L~(-1)或10 μmol·L~(-1)的PF進行榦預.免疫組化法計數黑質緻密部酪氨痠羥化酶(tyrosine hydroxylase,TH)暘性細胞數,定量RT-PCR法檢測α-syn mRNA的錶達豐度.結果 MPP~+(0.1、0.5、1.0 mmol·L~(-1))處理導緻黑質緻密部TH+細胞數較正常對照組減少,差異均有統計學意義(P<0.05或P<0.01);MPP~+(0.5 mmol·L~(-1))使α-syn mRNA的錶達水平明顯升高(P<0.01).PF(10 μmol·L~(-1))有效增加瞭TH+細胞的數量(P<0.01),併明顯下調瞭α-syn mRNA的錶達(P<0.05).結論 PF能有效拮抗MPP~+導緻的多巴胺能神經元受損死亡,其機製可能與下調α-syn的錶達有關.
목적 연구작약감(paeoniflorin,PF)대MPP~+소치대서흑질뇌편다파알능신경원손상적보호작용,병관찰PF대α-synuclein(α-syn) mRNA표체수평적영향.방법 리체배양SD대서흑질기관형뇌편,배양적d 10급여불동농도적MPP~+(0.1、0.5、1.0 mmol·L~(-1))처리24 h,병재0.5 mmol·L~(-1) MPP~+처리적기출상,급여1 μmol·L~(-1)혹10 μmol·L~(-1)적PF진행간예.면역조화법계수흑질치밀부락안산간화매(tyrosine hydroxylase,TH)양성세포수,정량RT-PCR법검측α-syn mRNA적표체봉도.결과 MPP~+(0.1、0.5、1.0 mmol·L~(-1))처리도치흑질치밀부TH+세포수교정상대조조감소,차이균유통계학의의(P<0.05혹P<0.01);MPP~+(0.5 mmol·L~(-1))사α-syn mRNA적표체수평명현승고(P<0.01).PF(10 μmol·L~(-1))유효증가료TH+세포적수량(P<0.01),병명현하조료α-syn mRNA적표체(P<0.05).결론 PF능유효길항MPP~+도치적다파알능신경원수손사망,기궤제가능여하조α-syn적표체유관.
Aim To study the protective effects of Paeoniflorin (PF) on dopaminergic neurons in brain slice of substantia nigra treated with MPP~+ and to investigate the transcription of alpha-synuclei (α-syn) mRNA.Methods The organatypic brain slice culture of substantia nigra prepared from neonatal SD rats was placed on Millicell-CM porous membranes and cultured to day-10.Then the cultrues of slice were treated with different concentrations (0.1,0.5,1.0 mmol·L~(-1)) of MPP~+ for 24 h.Some of the cultrues treated with 0.5 mmol·L~(-1) MPP~+ also received PF (1 or 10 μmol·L~(-1)).Slices cultured in normal medium were used as vehicle control.The tyrosine hydroxylase (TH) immunohistochemical staining with the cell counting was used to determine the dopaminergic neruons.The transcription of α-syn mRNA was examined by real-time quantitative RT-PCR.Results MPP~+(0.1,0.5,1.0 mmol·L~(-1)) exposure markedly decreased the number of TH~+ cells in a dose-dependent manner (P<0.05 or P<0.01) and sharply induced the transcription of α-syn mRNA (P<0.01) in slices treated with 0.5 mmol·L~(-1) MPP~+.The addition of PF (10 μmol·L~(-1)) to MPP~+-treated slices significantly increased dopaminergic neurons survival (P<0.01) and downregulated the transcription of α-syn mRNA significantly (P<0.05).Conclusion PF can effectively inhibit the injury of dopaminergic neurons induced by MPP~+ on brain slice of substantia nigra and downregulate the transcription of alpha-synuclein.
