Danshensuan A regulates the effect of LncRNA TSPOAP1-AS1 on alleviating H2O2 induced apoptosis and angiogenesis in brain microvascular endothelial cells
Objective Exploring the effect of Danshensuan A regulating lncRNA TSPAOP1-AS1 on alleviat-ing H2O2 induced apoptosis and angiogenesis in brain microvascular endothelial cells.Methods A total of 80 pa-tients with ischemic stroke admitted to our hospital from January 2021 to March 2023 were selected as the study group,and an additional 80 healthy individuals who came to our hospital for health check-ups during the same peri-od were included as the control group.Logarithmic growth of HBMECs was divided into 9 groups:Control group,H2O2 group,SAA-L group,SAA-M group,SAA-H group,EDA group,si-TSPOAP1-AS1 group,si-NC group,and pcDNA-TSPOAP1-AS1 group.The relative expression levels of TSPOAP1-AS1 in serum and cells were detected us-ing RT-PCR.Cell viability was assessed by CCK-8 assay.Flow cytometry was used to detect cell apoptosis.Matrigel tube formation assay was performed to evaluate angiogenesis ability.ELISA was used to measure cellular oxidative stress.Western blotting was conducted to examine the protein expression of Caspase-3,Bax,and Bcl-2 in cells.Re-sults Compared to the control group(1.00±0.09),the relative expression level of TSPOAP1-AS1 in the serum of patients in the study group(4.36±0.37)was significantly increased(P＜0.01).Compared to the Control group,the H2O2 group showed significant increases in TSPOAP1-AS1 relative expression levels in cells(2.65±0.23),cell apop-tosis rate(18.29±1.65),MDA content in cells(40.81±3.72),and protein expression of Caspase-3(0.78±0.08)and Bax(0.82±0.09),as well as significant decreases in cell survival rate(35.36±3.65),tube formation(85.32±7.68),SOD activity in cells(15.48±1.63),GSH-Px content(9.43±0.82),and protein expression of Bcl-2(0.21±0.02)(P＜0.01).Compared with the H2O2 group,the relative expression levels of TSPOAP1-AS1 in cells of the SAA-H and si-TSPOAP1-AS1 groups(1.21±0.12)(1.30±0.13),apoptosis rate(5.31±0.50)(6.01±0.62),cellular MDA content(8.95±0.82)(9.39±0.92)and expression of Caspase-3 protein(0.32±0.04)(0.35±0.04)and Bax protein(0.40±0.05)(0.45±0.05)were significantly reduced,and the cell survival rate(87.21±7.64)(82.31±7.50),the number of tubule formation(198.75±18.32)(180.31±17.64),SOD activity(60.27±5.89)(55.26±5.24),GSH-Px content(40.63±4.17)(38.76±3.27),and Bcl-2 protein(0.70±0.08)(0.68±0.07)expression were significantly higher in the cells(P＜0.01).Compared to the SAA-H group,the pcDNA-TSPOAP1-AS1 group showed significant in-creases in TSPOAP1-AS1 relative expression levels in cells(2.49±0.20),cell apoptosis rate(16.90±1.60),MDA content in cells(39.01±3.83),and protein expression of Caspase-3(0.72±0.08)and Bax(0.78±0.08),as well as significant decreases in cell survival rate(40.47±2.89),tube formation(90.15±8.62),SOD activity in cells(20.16±2.11),GSH-Px content(11.56±1.08),and protein expression of Bcl-2(0.28±0.03)(P＜0.01).Conclu-sion Danshensuan A can promote the survival and angiogenesis of HBMECs induced by H2O2,inhibit cell apopto-sis,and alleviate oxidative stress response.Its mechanism of action may be related to the inhibition of lncRNA TSPOAP1-AS1.
Danshensuan ALong non-coding RNA(lncRNA)TSPOAP1-AS1Brain microvascu-lar endothelial cells(HBMECs)H2O2AngiogenesisApoptosis
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