中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2012年
9期
659-663
,共5页
栗永生%薛莉%韩珣%谢安木
慄永生%薛莉%韓珣%謝安木
률영생%설리%한순%사안목
Fas相关死亡结构域蛋白质%基因沉默%帕金森病%细胞凋亡
Fas相關死亡結構域蛋白質%基因沉默%帕金森病%細胞凋亡
Fas상관사망결구역단백질%기인침묵%파금삼병%세포조망
Fas-associated death domain protein% Gene silencing% Parkinsou disease%Apoptosis
目的 探讨Fas相关死亡结构域蛋白(FADD)基因沉默对帕金森病(PD)大鼠行为学变化及黑质死亡受体和半胱氨酸蛋白酶8(caspase-8)表达的影响.方法 应用化学修饰的干扰RNA及立体定向技术等,在制作PD模型前黑质内注入FADD siRNA、FADD siRNA阳性对照物或FADDsiRNA阴性对照物.观察正常对照组、PD组、FADD siRNA组、FADD siRNA阳性对照组和FADDsiRNA阴性对照组(每组各15只)大鼠阿朴吗啡诱导的旋转行为的改变；采用Western blot分析和RT-PCR等方法,检测以上5组大鼠黑质FADD、Fas和caspase-8蛋白及mRNA的半定量表达.结果 正常对照组大鼠阿朴吗啡诱导下无旋转行为,其余4组旋转次数超过6 r/min(持续30 min)的大鼠(只)分别为12(12/14)、3(3/13) 、4(4/15) 、11 (11/14),组间比较差异具有统计学意义(x2＝18.56,P=0.000)；与正常组比较,PD组各个蛋白及mRNA的表达均明显升高；与PD组比较,FADD siRNA组FADD和caspase-8蛋白及mRNA的表达受到明显抑制,差异具有统计学意义,但PD组和阴性对照组间及其余3组间比较差异均无统计学意义.与正常组比较,其余4组Fas蛋白及mRNA的表达均明显升高,差异具有统计学意义,但4组间比较差异无统计学意义.结论 死亡受体凋亡通路在PD发病机制中起重要作用,而FADDsiRNA能够有效抑制此通路,从而在一定程度上具有神经保护作用.
目的 探討Fas相關死亡結構域蛋白(FADD)基因沉默對帕金森病(PD)大鼠行為學變化及黑質死亡受體和半胱氨痠蛋白酶8(caspase-8)錶達的影響.方法 應用化學脩飾的榦擾RNA及立體定嚮技術等,在製作PD模型前黑質內註入FADD siRNA、FADD siRNA暘性對照物或FADDsiRNA陰性對照物.觀察正常對照組、PD組、FADD siRNA組、FADD siRNA暘性對照組和FADDsiRNA陰性對照組(每組各15隻)大鼠阿樸嗎啡誘導的鏇轉行為的改變；採用Western blot分析和RT-PCR等方法,檢測以上5組大鼠黑質FADD、Fas和caspase-8蛋白及mRNA的半定量錶達.結果 正常對照組大鼠阿樸嗎啡誘導下無鏇轉行為,其餘4組鏇轉次數超過6 r/min(持續30 min)的大鼠(隻)分彆為12(12/14)、3(3/13) 、4(4/15) 、11 (11/14),組間比較差異具有統計學意義(x2＝18.56,P=0.000)；與正常組比較,PD組各箇蛋白及mRNA的錶達均明顯升高；與PD組比較,FADD siRNA組FADD和caspase-8蛋白及mRNA的錶達受到明顯抑製,差異具有統計學意義,但PD組和陰性對照組間及其餘3組間比較差異均無統計學意義.與正常組比較,其餘4組Fas蛋白及mRNA的錶達均明顯升高,差異具有統計學意義,但4組間比較差異無統計學意義.結論 死亡受體凋亡通路在PD髮病機製中起重要作用,而FADDsiRNA能夠有效抑製此通路,從而在一定程度上具有神經保護作用.
목적 탐토Fas상관사망결구역단백(FADD)기인침묵대파금삼병(PD)대서행위학변화급흑질사망수체화반광안산단백매8(caspase-8)표체적영향.방법 응용화학수식적간우RNA급입체정향기술등,재제작PD모형전흑질내주입FADD siRNA、FADD siRNA양성대조물혹FADDsiRNA음성대조물.관찰정상대조조、PD조、FADD siRNA조、FADD siRNA양성대조조화FADDsiRNA음성대조조(매조각15지)대서아박마배유도적선전행위적개변；채용Western blot분석화RT-PCR등방법,검측이상5조대서흑질FADD、Fas화caspase-8단백급mRNA적반정량표체.결과 정상대조조대서아박마배유도하무선전행위,기여4조선전차수초과6 r/min(지속30 min)적대서(지)분별위12(12/14)、3(3/13) 、4(4/15) 、11 (11/14),조간비교차이구유통계학의의(x2＝18.56,P=0.000)；여정상조비교,PD조각개단백급mRNA적표체균명현승고；여PD조비교,FADD siRNA조FADD화caspase-8단백급mRNA적표체수도명현억제,차이구유통계학의의,단PD조화음성대조조간급기여3조간비교차이균무통계학의의.여정상조비교,기여4조Fas단백급mRNA적표체균명현승고,차이구유통계학의의,단4조간비교차이무통계학의의.결론 사망수체조망통로재PD발병궤제중기중요작용,이FADDsiRNA능구유효억제차통로,종이재일정정도상구유신경보호작용.
Objective To investigate the effects of gene silencing of Fas-associated death domain (FADD) with synthetic small interfering RNA (siRNA) on apomorphine-induced contralateral rotation,and the expression of Fas and caspase-8 in rat models of parkinsonism. Methods Sprague-Dawley rats were randomly divided into 5 groups:control group,Parkinson' s disease (PD) group,FADD siRNA group,FADD siRNA positive control group and FADD siRNA negative control group. Synthetic FADD siRNA sequences,siRNA positive sequences or siRNA negative sequences were infused into right substantianigra of midbrain using RNA interference and stereotactic techniques before parkinsonian rat model establishment.Apomorphine-induced contralateral rotations of the rats were observed after the injection.The protein and mRNA expression levels of FADD,Fas and caspase-8 were measured by Western blot and RT-PCR.Results In the control group,no rotation was observed after injecting apomorphine; however,in the rest groups,the number of rats respectively was 12 (12/14),3 (3/13),4 (4/15) and 11 ( 11/14 ) in apomorphine-induced contralateral rotation,which had significant statistical differences ( x2 ＝ 18.56,P =0.000).In parkinsonian substantia nigra,marked increases in the protein and mRNA levels of FADD,Fas and caspase-8 were observed,compared with control group.Furthermore,compared with PD group,FADD protein and mRNA levels were strongly suppressed by administration of FADD siRNA in FADD siRNA group. FADD siRNA administration also resulted in great attenuation of 6-hydroxydopamine-induced increases in expression and activation of caspase-8.However,no decrease in expression of Fas was observed in FADD siRNA group and FADD siRNA positive control group,compared with PD group.Conclusion Our results suggest that death receptor signaling pathway plays a critical role in the pathogenesis of PD.FADD siRNA is effective against this pathway and it may,at least in part,provide a potential neuroprotective effect.
