Background:Mesencephalic astrocyte-derived neurotrophic factor(MANF)is a new candidate growth factor for dopaminergic neurons against endoplasmic reticulum stress(ER stress).HSP70 family,a chaperon like heat shock pro...Background:Mesencephalic astrocyte-derived neurotrophic factor(MANF)is a new candidate growth factor for dopaminergic neurons against endoplasmic reticulum stress(ER stress).HSP70 family,a chaperon like heat shock protein family,was proved to be involved in the MANF induced survival pathway in 6-OHDA treated SHSY-5Y cells.However,the ER stress relative transcriptome,in MANF signaling cascades is still investigated.The involvement of HSP70,a 70kd member of HSP70 family,need further to be verified.Methods:The cell apoptosis was assayed by MTT,TUNEL staining and western blot of cleaved Caspase-3.The differentially expressed genes in SHSY-5Y cells under different conditions(control,6-OHDA,6-OHDA+MANF)were investigated by RNA-seq.Expression of HSP70 was further confirmed by real-time PCR.RNAi knockdown for HSP70 was performed to investigate the role of HSP70 in the MANF signaling pathway.Results:MANF inhibits 6-OHDA-induced apoptosis in SHSY-5Y cells.Six ER stress relative genes(HSP70,GRP78,xbp-1,ATF-4,ATF-6,MAPK)were found enriched in 6-OHDA+MANF treatment group.HSP70 was the most significantly up-regulated gene under 6-OHDA+MANF treatment in SHSY-5Y cells.RNAi knockdown for HSP70 inhibits the protective effects of MANF against 6-OHDA toxicity in SHSY-5Y cells.Conclusion:MANF exerts a protective role against 6-OHDA induced apoptosis in SHSY-5Y cells via up-regulating some ER stress genes,including HSP70 family members.The HSP70 expression level plays a key role in MANFmediated survival pathway.展开更多
AIM: To determine whether the use of an antioxidant reagent coenzyme Q10 can protect human dopaminergic cell line SHSY-5Y against rotenone-induced apoptosis. METHODS: Cell viability was quantified by 3-(4,5-dimethylth...AIM: To determine whether the use of an antioxidant reagent coenzyme Q10 can protect human dopaminergic cell line SHSY-5Y against rotenone-induced apoptosis. METHODS: Cell viability was quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the apoptosis induced by rotenone was observed by Hoechst 33342, propidium iodide, and calcein AM staining in SHSY-5Y cells. RESULTS: Rotenone, a commonly used natural pesticide, inhibitor of mitochondrial respiratory chain complex I, was shown to decrease cell viability and induce apoptosis in SHSY-5Y cells. Pretreated with coenzyme Q10, the electron transporter in the mitochondrial respiratory chain, remarkably increased cell viability as well as significantly reduced the percentage of apoptotic SHSY5Y cells induced by rotenone. CONCLUSION: Coenzyme Q10 has a beneficial effect in protecting against rotenone-induced apoptosis in SHSY-5Y cells. The results support the hypothesis that coenzyme Q10 is a component of the antioxidant machinery that protects cell membranes from oxidative damage and decreases apoptosis. Further evaluation is required to determine whether the neuroprotective action of coenzyme Q10 can be used for the prevention and therapy of neurodegenerative diseases.展开更多
文摘目的研究姜黄素(Curcumin)对于血红素加氧酶-1(heme oxygenase-1,HO-1)和核因子相关因子2(nuclear factor erythroid2-related factor2,Nrf-2)表达的影响,探讨Curcumin保护神经细胞的作用机制。方法 SH-SY5Y(人神经母细胞瘤)细胞用0、1.25、5.0、20.0μmol·L-1 Curcumin处理24h,以及用5.0μmol·L-1 Curcumin分别处理细胞0、12、24、48h,然后用RT-PCR和Western blot检测各组的mRNA和蛋白的表达,并在各个浓度组使用Nrf-2siRNA,Western blot检测转染Nrf-2siRNA后HO-1的表达情况。结果 Curcumin处理后,SH-SY5Y细胞中Nrf-2和HO-1的表达均明显增强,且呈时间和浓度依赖性(P<0.05),0、1.25、5.0、20.0μmol·L-1 Curcumin处理的细胞24h后HO-1表达与用0,1.25,5.0,20.0μmol·L-1 Curcumin和转染Nrf-2siRNA同时处理24h后的细胞HO-1表达相比明显下降(group 1 vs group 5,group 2 vs group 6,group 3 vsgroup 7,group 4 vs group 8,P<0.05)。结论 Curcumin通过Nrf-2诱导HO-1的表达作用,可能是其抑制神经细胞氧化应激、发挥保护作用的机制之一。
基金This study is supported by the National Major Scientific and Technological Special Project for Significant New Drugs Development(2014ZX09102043-003)National Natural Science Foundation of China(81371403)+1 种基金Shanghai Science and Technology Commission(13JC1401102)Natural Science Foundation of Jiangsu Province of China(BK20140275).
文摘Background:Mesencephalic astrocyte-derived neurotrophic factor(MANF)is a new candidate growth factor for dopaminergic neurons against endoplasmic reticulum stress(ER stress).HSP70 family,a chaperon like heat shock protein family,was proved to be involved in the MANF induced survival pathway in 6-OHDA treated SHSY-5Y cells.However,the ER stress relative transcriptome,in MANF signaling cascades is still investigated.The involvement of HSP70,a 70kd member of HSP70 family,need further to be verified.Methods:The cell apoptosis was assayed by MTT,TUNEL staining and western blot of cleaved Caspase-3.The differentially expressed genes in SHSY-5Y cells under different conditions(control,6-OHDA,6-OHDA+MANF)were investigated by RNA-seq.Expression of HSP70 was further confirmed by real-time PCR.RNAi knockdown for HSP70 was performed to investigate the role of HSP70 in the MANF signaling pathway.Results:MANF inhibits 6-OHDA-induced apoptosis in SHSY-5Y cells.Six ER stress relative genes(HSP70,GRP78,xbp-1,ATF-4,ATF-6,MAPK)were found enriched in 6-OHDA+MANF treatment group.HSP70 was the most significantly up-regulated gene under 6-OHDA+MANF treatment in SHSY-5Y cells.RNAi knockdown for HSP70 inhibits the protective effects of MANF against 6-OHDA toxicity in SHSY-5Y cells.Conclusion:MANF exerts a protective role against 6-OHDA induced apoptosis in SHSY-5Y cells via up-regulating some ER stress genes,including HSP70 family members.The HSP70 expression level plays a key role in MANFmediated survival pathway.
基金Supported by foundation for build-up of key disciplines from the Education Department of Hunan province .
文摘AIM: To determine whether the use of an antioxidant reagent coenzyme Q10 can protect human dopaminergic cell line SHSY-5Y against rotenone-induced apoptosis. METHODS: Cell viability was quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the apoptosis induced by rotenone was observed by Hoechst 33342, propidium iodide, and calcein AM staining in SHSY-5Y cells. RESULTS: Rotenone, a commonly used natural pesticide, inhibitor of mitochondrial respiratory chain complex I, was shown to decrease cell viability and induce apoptosis in SHSY-5Y cells. Pretreated with coenzyme Q10, the electron transporter in the mitochondrial respiratory chain, remarkably increased cell viability as well as significantly reduced the percentage of apoptotic SHSY5Y cells induced by rotenone. CONCLUSION: Coenzyme Q10 has a beneficial effect in protecting against rotenone-induced apoptosis in SHSY-5Y cells. The results support the hypothesis that coenzyme Q10 is a component of the antioxidant machinery that protects cell membranes from oxidative damage and decreases apoptosis. Further evaluation is required to determine whether the neuroprotective action of coenzyme Q10 can be used for the prevention and therapy of neurodegenerative diseases.
