Effects of Cytochalasin D on Expression of Aquaporins and Inward Rectifying Potassium Channel 4.1 Gene in Spinal Cord Astrocytes of Rats / 中国康复理论与实践

ABSTRACT

@#Objective To investigate the expression of aquaporin (AQP) 1, AQP4, inward rectifying potassium channel 4.1 (Kir4.1) and cytoskeleton features of rat spinal cord astrocytes after cytochalasin D (CytD) intervention. Methods Spinal cord astrocytes isolated from 2~3-day-old rats were cultured till confluency. MTT was used to assess survival rate of astrocytes 2 h, 12 h and 24 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD, respectively. Confocal microscopy was used to observe cytoskeleton features of astrocytes 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml of CytD. The expression of AQP1, AQP4, Kir4.1 mRNA were determined with real-time PCR 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD. Results The survival rate of rat spinal cord astrocytes reduced with the time of co-culture and concentration of CytD (P<0.05). The cytoskeleton of astrocytes was reconstructed. The expression of AQP1, AQP4 and Kir4.1 mRNA increased after co- cultured with 0.05~0.40 μg/ml of CytD. Conclusion The appropriate dosage of CytD may remodel the cytoskeleton and increase the mRNA expression of AQP1, AQP4 and Kir4.1 in spinal cord astrocytes of rats.