结蛋白介导的雌激素受体对颅内动脉瘤血管平滑肌 细胞表型的调控作用.

隋昕; 隋天意; 张旭; 冯旭; 王东玉

Objective To investigate desmin expression in intracranial aneurysm tissue and its mediating effect on the phenotype regulation of vascular smooth muscle cell (VSMC) by estrogen receptor (ER). Methods Twenty-eight samples of surgical site specimens including external carotid artery branches and intracranial aneurysm resection were collected from patients with nonarterial aneurysms in clinical practice. Desmin expression in the arterial blood vessels and intracranial aneurysms was detected using immunohistochemical staining (SABC), whereas desmin expression in the intracranial aneurysm tissue and normal blood vessels was detected using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blotting. The VSMC were divided into control, shNC, shDesmin, pcDNA3.1/NC, pcDNA3.1/Desmin, and pcDNA3.1/Desmin 3-MA groups. The shNC, shDesmin, pcDNA3.1/NC, and pcDNA3.1/ Desmin groups were transfected with shNC, shDesmin, pcDNA3.1/NC, or pcDNA3.1/Desmin plasmids, respectively. The pcDNA3.1/ Desmin 3-MA group was pretreated with 10 μmol/L 3-MA for 6 hours before transfection was performed. qRT-PCR and Western blotting were used to detect the expression of silenced DES in VSMC. CCK-8 assay was used to detect the effect of silencing DES on the proliferation ability of VSMC. Transwell cell migration assay was used to detect the effect of silencing DES on the migration ability of VSMC. Western blotting was used to detect the effects of DES silencing on the ERα/ERβ ratio, and phenotype regulatory protein levels in VSMC. Results Immunohistochemical staining showed that the positive expression rate of desmin in intracranial aneurysms was significantly lower than that in normal arterial blood vessels (χ² = 16.601, P < 0.001) . Compared with normal vascular tissue, desmin expression in intracranial aneurysm tissue was reduced (P < 0.001) . Compared with the shNC group, the expression level of desmin in VSMC of the shDesmin group decreased (P < 0.001), the cell proliferation was stronger (P < 0.05), the number of cell migrations increased (P < 0.001), and the ERα/ERβ protein expression ratio increased (P < 0.001) . Compared with the pcDNA3.1/NC group, the expression levels of human smooth muscle α-actin (SM-α-actin) and human smooth muscle myosin heavy chain (SM-MHC) proteins in the VSMC of the pcDNA3.1/ Desmin group increased (P < 0.01), while the expression levels of matrix metalloproteinase-3 (MMP-3) and tumor necrosis factor-α (TNF-α) proteins decreased (P < 0.05) . When compared with the pcDNA3.1/Desmin group, the expression levels of SM-α-actin and SM-MHC proteins in the VSMC of the pcDNA3.1/Desmin 3-MA group decreased (P < 0.05), while the expression levels of MMP-3 and TNF-α proteins increased (P < 0.05) . Conclusion Desmin is expressed at low levels in intracranial aneurysm tissues. Silencing DES promotes VSMC proliferation and migration, which may, in turn, promote VSMC phenotype regulation.

VASCULAR smooth muscle; INTRACRANIAL aneurysms; IMMUNOSTAINING; MATRIX metalloproteinases; ESTROGEN receptors

Journal of China Medical University, 2024, Vol 53, Issue 12, p1094

0258-4646

Academic Journal

10.12007/j.issn.0258-4646.2024.12.006