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Neuroprotective Effects of Salidroside in the PC12 Cell Model Exposed to Hypoglycemia and Serum Limitation

Author: YuShu
Tutor: DingZuo；LiuMei
School: Nantong University
Course: Neurobiology
Keywords: PC12 cells Salidroside Low sugar low serum injury Apoptosis Protection of neurons
CLC: R285.5
Type: Master's thesis
Year: 2008
Downloads: 126
Quote: 0
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Abstract


Objective To observe the salidroside (Salidroside) the protective effect of low sugar low serum induced damage in PC12 cells. Method 1, the in vitro culture of PC12 cells, low-sugar low serum injury model using low sugar low serum medium preparation PC12 cells. By MTT assay, observed after low sugar low serum injury time (12h, 24h, 36h, 48h) PC12 cell viability changes. 2, in vitro cultured PC12 cells treated with different concentrations of salidroside (80μg/ml, 160μg/ml, 320μg/ml) pre-protection 24h after low sugar low serum injury 24h, morphological analysis and MTT assay, observing the Red King days glycosides low sugar low serum injury PC12 cell morphology and vitality. By Hoechst33342 staining and flow cytometry analysis were observed of salidroside on low sugar low serum injury apoptosis in PC12 cells. 4, by RT-PCR and Western blot, low sugar low serum injury in PC12 cells Bcl-2 and Bax mRNA and its protein expression observed salidroside. The 5, salidroside low sugar low serum injury of PC12 cells Caspase3 mRNA and its protein activity activity was detected by RT-PCR and Caspase3. The 6, lipophilic cationic fluorescent dye Rhodamine123 marked PC12 cells by laser scanning confocal microscopy to detect low sugar low serum stimulation of mitochondrial transmembrane potential changes in PC12 cells. Observe the salidroside pre protection of low sugar low serum injury PC12 cell mitochondrial transmembrane potential. 7, the use of molecular probes DCFH-DA labeled PC12 cells fluorescent product DCF fluorescence intensity was measured by flow cytometry, to observe the the salidroside pre protection of low sugar low serum induced damage in PC12 cells reactive oxygen species (ROS) content impact. 1, MTT results show low sugar low serum cultured obvious cytotoxicity in PC12 cells, cell viability decreased with low sugar low serum incubation time. Morphological observation and MTT assay results showed that salidroside antagonized low sugar low serum induced damage in PC12 cell viability decreased, along with the increase of the concentration of the drug in the dose range of the observed role gradually enhanced, 320μg/ml role to the most significant (P lt; 0.05). 3, Hoechst 33342 staining and flow cytometry results show a significant reduction in the low-sugar low serum induced damage in PC12 cells apoptosis (P lt; 0.05) salidroside. RT-PCR and Western blot results suggest that salidroside antagonistic of Bcl-2/Bax mRNA protein ratio in the low sugar low serum damage caused PC12 cells decreased, this effect is a dose-response relationship, to 320μg/ml The role of the most significant (P lt; 0.05). RT-PCR and Caspase3 activity salidroside antagonized low sugar low serum induced damage in PC12 cells Caspase3 mRNA upregulation and increased activity of the protein, a dose-effect relationship of this role and the to 320μg/ml role of the most significant (P lt; 0.05). 6, mitochondrial transmembrane potential test results show low sugar low serum damage, PC12 cells decreased mitochondrial transmembrane potential salidroside pre protection dropped significantly antagonize the mitochondrial transmembrane potential (P lt; 0.05). 7, reactive oxygen species (ROS), test results show low sugar low serum injury in PC12 cells ROS were significantly increased, while salidroside pre protection can significantly antagonize intracellular ROS levels increased (P lt; 0.05) , play a clear role of ROS. Conclusions salidroside antagonized low sugar low serum PC12 cells injury, and this effect showed a dose-related. 2, the protective effect of salidroside on low sugar low serum injury PC12 cells and their regulation of apoptosis-related genes, to stabilize mitochondrial transmembrane potential and clear intracellular ROS related.

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