AI Article Synopsis
- The study aimed to investigate how recombinant adenovirus carrying PDE5-targeting shRNAs affects calcium levels in rat penile smooth muscle cells, with potential applications in treating erectile dysfunction (ED).
- Researchers transfected rat smooth muscle cells with specific shRNAs and measured calcium activity using a fluorescent probe over 24, 48, and 72 hours, observing the changes through a laser scanning confocal microscope.
- Results showed that the intracellular calcium levels in the shRNA group were significantly lower compared to control groups, indicating that adenovirus-mediated shRNAs can effectively reduce calcium levels in penile smooth muscle cells, suggesting a promising gene therapy approach for ED.
Article Abstract
Objective: To observe the influence of recombinant adenovirus-mediated PDE5-shRNAs on the free calcium level in rat penile smooth muscle cells and to explore the feasibility of gene therapy for erectile dysfunction (ED).
Methods: Smooth muscle cells of the rat corpus cavernosum were transfected with constructed rAd-rPDE5-shRNAs and then dyed with the calcium fluorescent probe Fluo-3/AM at 24, 48 and 72 hours. The dynamic changes of the calcium fluorescence intensity were observed under the laser scanning confocal microscope (LSCM). The relative level of intracellular calcium was determined by fluorescence indexes.
Results: The fluorescence indexes of calcium at 24, 48 and 72 hours were 829.3 +/- 7.8, 801.5 +/- 9.5 and 856.3 +/- 8.7 in the rAd-rPDES-shRNAs group, significantly lower than in the rAd-mock (1106.3 +/- 10.8, 1121.3 +/- 10.2 and 1058.5 +/- 12.1) and blank control group (1076.6 +/- 9.7, 1133.4 +/- 11.2 and 1104.3 +/- 10.5) (P < 0.05).
Conclusion: Adenovirus mediated shRNAs of the target PDE5 gene can significantly decrease the intracellular calcium level in the smooth muscle cells of the rat corpus cavernosum.
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