左卡尼汀通過內質網(wǎng)途徑抑制H2O2誘導的SH-SY5Y細胞損傷的研究.pdf

1、分類號：R966 密級：不保密UDC：610 學校代碼：11065碩士學位論文左卡尼汀通過內質網(wǎng)途徑抑制 左卡尼汀通過內質網(wǎng)途徑抑制 H2O2 誘導 誘導的SH-SY5Y 細胞損傷的研究 細胞損傷的研究研 究 生 王沖指 導 教 師 曹玉 韓彥弢學科專業(yè)名稱 藥理學論文答辯日期 2016 年 11 月 26 日2ABSTRACTOBJECTIVE To copy hydrogen peroxide (H2O2)-induced oxid

2、ative damageAlzheimer's disease model in SH-SY5Y cells, and explore the protective effect ofL-carnitine in SH-SY5Y cells via modulating endoplasmic reticulum apoptotic pathway.METHODS The cells were divided into grou

3、ps randomly as below: control group,H2O2 (400µmol·L-1 ) model group, L-carnitine pretreated group (50、100、200 µmol·L-13h pretreatment followed by H2O2 treatment) and 4-PBA pretreated group (5µmol

4、·L-1 3hpretreatment followed by H2O2 treatment). DNA ladder and Annexin V-FITC stainingwere preformed to identify the apoptosis in SH-SY5Y cells. Generation of ROS wasdetected by 2’, 7’-Dichlorofluorescin diacetate

5、(DCFH-DA). RT-PCR was preformed todetect the transcription of GRP78. The expression of GRP78, CHOP and caspase-12 weredetermined by western-blot. RESULTS The cells were incubated with 400µmol·L-1H2O2 for 20 min

6、 to establish the oxidative damage model. Obvious DNA ladder wasobserved in H2O2 treated group and the ladders in L-carnitine pretreated groups weremuch indistinct. The result of annexin V FITC/PI staining showed that th

7、e apoptosis inthe early stage was inhibited as well as in the late stage of apoptosis and necrosis inL-carnitine pretreated groups compared with the H2O2 treated group. The results ofRT-PCR and western-blot indicated the

8、 transcription and expression of GRP78 wereincreased in the H2O2 treated group and decreased in the L-carnitine pretreated groups.Compared with the H2O2 treated group, the expression of CHOP and caspase-12 weremuch lower

9、 in the L-carnitine pretreated groups. The result of DCFH-DA probe indicatedthat ROS was over-generated after H2O2 treatment, and the L-carnitine pretreatment couldobviously inhibit the ROS generation. 4-PBA could yield

10、a week inhibitory effect to theROS generation induced by H2O2 treatment.CONCLUSION L-carnitine could inhibitH2O2 induced apoptosis in SH-SY5Y cells by down-regulating of the transcription andexpression of GRP78, and decr

11、easing the expression of CHOP and caspase-12. Theprotective effect of L-carnitine was related to the inhibition of ROS generation inendoplasmic reticulum.Postgraduate student: Blind EvaluationDirected by Prof: Blind Eval