Hypoxia-induced inflammatory response in penile corpora cavernosal smooth muscle cells of sd rats and the interventional roleof lycopen
ZHANG Xin,FU Yi-jia,HUANG Wen-jie
The Second School of Clinical Medicine，Zhejiang University of Chinese Medicine，Hangzhou，Zhejiang (310053)，China
Abstract:
Objective To explore the hypoxia-induced inflammatory response in corpus cavernosum smooth muscle cells (CCSMCs) of SD rats and the regulatory effect of lycopene. Methods Primary CCSMCs were isolated from SD rats and observed for morphological stability up to the fourth passage. Immunofluorescence staining for α-smooth muscle actin (α-SMA) and desmin was performed to verify the purity of cultured CCSMCs. After 24 hours of exposure to different concentrations of CoCl2 and lycopene, either alone or in combination, cell viability was assessed using the Cell Counting Kit-8 (CCK8). The experiment included three treatment groups: control, hypoxia (200 μmol/LCoCl2), and lycopene treatment (40 μmol/L). Western Blotting and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used to detect the levels of hypoxia-inducible factor 1-alpha (HIF-1α), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-18 (IL-18), and interleukin-1 beta (IL-1β) after 24 hours of culture. Results The CCSMCs maintained morphological uniformity up to the fourth passage. High purity was confirmed through positive staining for α-SMA and desmin. Following 200 μmol/L CoCl2 treatment, CCSMCs viability decreased to 50%, with increases in intracellular levels of HIF-1α, TNF-α, IL-6, IL-18 (P<0.05), and a significant rise in IL-1β mRNA expression (P<0.05), though protein levels showed only an increasing trend (P>0.05). Lycopene at 40 μmol/L elevated CCSMC viability to 80% of normal levels and significantly reduced HIF-1α, TNF-α, IL-18 protein expression, and TNF-α, IL-18, IL-1β mRNA levels (P<0.05). IL-6 protein, IL-6 mRNA, and IL-1β protein levels showed a decreasing trend (P>0.05). Conclusion Under hypoxic conditions, the expression of inflammatory factors in CCSMCs is upregulated, and lycopene can effectively inhibit the expression of these inflammatory factors.
