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Objective To analyze the protective effects ofbutylphthalide(NBP) on apoptosis factors (p-JNK,Fas and FasL) of death receptor pathway in JNK pathway of cell model of Parkinson's disease (PD).Methods SH-SY5Y cell apoptosis model induced by MPP + was established in vitro.The cells were divided into four groups:normal control group,SHSYSY cells were treated with complete medium without drug intervention;MPP+ group,1 mmol·L-1 MPP+ was added into the cells;NBP+ MPP+ group,the cells were pretreated with 10 mol·L-1 NBP for 3 h and added with 1 mmol·L-1 MPP+;SP600125 + MPP+ group,the cells were cultured with 10 mol·L-1 JNK inhibitor SP600125 pretreatment for 3 h and 1 mmol·L-1 MPP+ was added.The proliferative potentiality of SH-SY5Y cells induced by MPP+ was measured by MTT.The apoptotic rate was analyzed by Annexin-V/PI (FCM).The morphology of SH-SY5Y cells was observed by inverted phase contrast microscope.The expression of apoptotic protein p-JNK,Fas,FasL was detected by Western blotting.Results The cell proliferative potentiality in the MPP+ group (49.30 ± 2.07)％ was significantly lower than that of the normal control group (100.00 ±0.00)％ (P ＜ 0.05).The cell proliferative potentiality in NBP + MPP + group and SP600125 + MPP + group were (71.90 ±2.10) ％ and (76.40 ± 2.80) ％,which was significantly higher than that of the MPP + group (P ＜ 0.05).Apoptosis rate in the MPP + group (32.27 ± 2.26) ％ was significantly higher than that of the normal control group (10.63 ± 2.07) ％ (P ＜ 0.05).The apoptosis rate in the NBP + MPP + group and SP600125 + MPP+ group were (21.13 ± 3.63) ％ and (19.15 ± 2.63) ％,and the apoptosis rate was significantly lower than that in the MPP+ group(P ＜0.05).The protein expression levels of p-JNK,Fas and FasL were significandy lower in NBP + MPP+ group and SP600125 + MPP+ group than that in the MPP+ group (P ＜0.05).Conclusion Butylphthalide can protect the injury of SH-SY5Y cells induced by MPP+.The mechanism of butylphthalide inhibiting apoptosis may be achieved through regulating p-JNK,Fas and FasL protein expression of death receptor pathway in JNK pathway and inhibiting the cell apoptosis.
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Objective To observe the effects of butylphthalide on the expression of autophagy-related protein and mRNA in l-meth-yl-4-phenyl-pyridiniumion (MPP+)-induced SH-SY5Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson's disease. Methods The SH-SY5Y cells were divided into control group (A), MPP+group (B), rapamycin pre-treated+MPP+group (C) and Butylphthalide pretreated+MPP+group (D). The relative viability of SH-SY5Y cells induced by MPP+was measured with MTT assay, the morphology of SH-SY5Y cells was observed. The expression of microtubule associated protein 1 light chain 3 (LC3)-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 mRNA were assayed by re-al-time quantitative reverse transcription-PCR (RT-PCR). Results The viability rates of cells were significantly lower in group B than in group A (t=20.270, P8.770, P6.647, P3.630, P<0.01), however, there was no significantly differ-ence between groups C and D (t<2.238, P≥0.05). Conclusion Butylphthalide could prevent the injury of SH-SY5Y cells induced by MPP+, which may affect Parkinson's disease by inducing autophagy.
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@#Objective To explore the effects of butylphthalide on the apoptosis of SH-SY5Y cells induced by the 1-methyl-4-phenylpyridinium iodide (MPP+). Methods SH-SY5Y cells were cultured with butylphthalide in the dosage of 1 µmol/L, 10 µmol/L and 20 µmol/L for 2 hours, and with MPP+ for 24 hours. The viability of cells was measured with trypan blue. The mitochondrial membrane potential was detected with Rhodamine 123. The content of P53 protein was detected with ELISA. The expression of P53 and cytochrome C (CytC) were detected with Western blotting. Results The viability of SH-SY5Y cells increased with the dosage of butylphthalide (P<0.05), while the mitochondrial membrane potential and the expression of P53 and CytC decreased (P<0.05). Conclusion Butylphthalide can prevents the SH-SY5Y cells from the injury induced by MPP+, which may associate with the inhibition of apoptosis through the expression of P53 and CytC.
ABSTRACT
Objective To explore the effects of butylphthalide on the apoptosis of SH-SY5Y cells induced by the 1-methyl-4-phenylpyri-dinium iodide (MPP+). Methods SH-SY5Y cells were cultured with butylphthalide in the dosage of 1 μmol/L, 10 μmol/L and 20 μmol/L for 2 hours, and with MPP+for 24 hours. The viability of cells was measured with trypan blue. The mitochondrial membrane potential was de-tected with Rhodamine 123. The content of P53 protein was detected with ELISA. The expression of P53 and cytochrome C (CytC) were de-tected with Western blotting. Results The viability of SH-SY5Y cells increased with the dosage of butylphthalide (P<0.05), while the mito-chondrial membrane potential and the expression of P53 and CytC decreased (P<0.05). Conclusion Butylphthalide can prevents the SH-SY5Y cells from the injury induced by MPP+, which may associate with the inhibition of apoptosis through the expression of P53 and CytC.
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Proteome research has become a new hot spot in the post-genome era. High-resolution two-dimensional gel electrophoresis (2-DE), which provides the most comprehensive analysis system of the whole proteome, was highly improved in recent years. With the development of computer techniques, the powerful and user-friendly image analysis systems appeared to help high-throughput, large-scale proteomic studies. Using new generation two-dimensional image analysis software, ImageMaster 2D Elite, the 2D gels of proteins extracted from cultured Schwann’s cells were processed. The analysis procedure, including image acquirement, spot detection, match, background subtraction, pI/Mr calibration, analysis results report and database query, were reported and discussed.