Effects of gene expression of aquaporins-4 and K+ ion channel protein-4.1 after cytoskeleton reconstruction by jasplakinolide in rat spinal cord astrocytes

English Keywords:Astrocytes  Microfilament  Proliferation

English Abstract:

  【Abstract】 Objectives: To investigate the mRNA expression of aquaporins-4(AQP4) and potassium ion channel protein-4.1(Kir4.1) by different concentration of jasplakinolide(JSK), a microfilament polymerization agent, in spinal cord astrocytes of rats. Methods: The primary spinal cord astrocytes from new born rats were extracted and cultured. 3-4, 5-Dimethylthiazol-2-yl-25-diphenyltetrazolium brom(MTT) was used to measure the cytotoxicity of JSK by different concentrations(0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml). The changes of cytoskeleton were monitored under the Laser Scanning Confocal Microscopy(LSCM). The mRNA levels of AQP4 and Kir4.1 were measured after JSK with different concentrations by real-time PCR. Results: MTT assay showed no significant difference in 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml groups compared with control group at 2h. LSCM showed that JSK of 0.05μg/m, 0.10μg/ml and 0.20μg/ml could remodel the spinal cord astrocyte cytoskeleton, nevertheless, 0.40μg/ml could damage cytoskeleton. Real-time PCR analysis showed that JSK of 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml concentrations could significantly decrease the gene expression of AQP4 and Kir4.1, which was statistically significant compared with control group(P<0.05). Furthermore, there were significant differences among 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml groups. Conclusion: JSK of 0.05μg/ml, 0.10μg/ml and 0.20μg/ml can remodel the cytoskeleton and decrease the mRNA expression of AQP4 and Kir4.1.