| 高寒冰,张开创,黄林轲,等.TRPC6基因对过氧化氢诱导大鼠脑血管平滑肌细胞炎症反应、氧化应激损伤的影响[J].安徽医药,2025,29(3):541-545. |
| TRPC6基因对过氧化氢诱导大鼠脑血管平滑肌细胞炎症反应、氧化应激损伤的影响 |
| Effect of TRPC6 gene on hydrogen peroxide induced inflammatory response and oxidative stress damage in rat cerebral vascular smooth muscle cells |
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| DOI：10.3969/j.issn.1009-6469.2025.03.021 |
| 中文关键词: 颅内动脉瘤 脑血管平滑肌细胞 瞬时受体电位阳离子通道蛋白 6(TRPC6)基因 炎症反应 氧化应激 亡，可， |
| 英文关键词: Intracranial aneurysm Cerebrovascular smooth muscle cell Transient receptor potential cationic channel protein 6 (TRPC6) gene Inflammatory response Oxidative stress |
| 基金项目:2019年度河南省医学科技攻关计划联合共建项目( LHGJ20190592) |
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| 中文摘要: |
| 目的探究瞬时受体电位阳离子通道蛋白 6(TRPC6)基因对过氧化氢(H2O2)诱导大鼠脑血管平滑肌细胞( VSMCs)损伤的影响。方法于 2022年 3—12月完成研究。分离大鼠脑基底动脉 VSMCs，采用随机数字表法分为五组:空白组、 H2O2组(500 μmol/L H2O2处理 6h)、 H2O2+空转染组( 500 μmol/L H2O2处理 6h，并转染空载脂质体)、 H2O2+TRPC6 siRNA组( 500 μmol/L H2O2处理 6h，并转染 TRPC6 siRNA)和 H2O2+pcDNA3.1 TRPC6组( 500 μmol/L H2O2处理 6h，并转染 pcDNA3.1 TRPC6)。采用人胆囊收缩素 /缩胆囊素八肽(CCK-8)法检测细胞增殖活性，流式细胞术检测细胞凋亡，酶联免疫吸附测定检测细胞中白细胞介素(IL) -6、IL-1β、单核细胞趋化蛋白 1(MCP-1)、超氧化物歧化酶( SOD)、丙二醛， DCFH-DA荧光探针法检测细胞中活性氧水平。结果与 H2O2组比较， H2O2+TRPC6 siRNA组 D(λ)450 nm值明显升高，细胞凋亡率明显降低( P<0.05)H2O2+pcDNA3.1 TRPC6组 D(λ)450 nm值明显降低，细胞凋亡率明显升高(P<0.05)。 H2O2组、 H2O2+TRPC6 siRNA组、 H2O2+pcDNA3.1，TRPC6组细胞中 IL-6含量分别为 |
| 英文摘要: |
| Objective To explore the effects of transient receptor potential cationic channel protein 6 (TRPC6) gene on hydrogen per-oxide (H2O2)-induced cerebrovascular smooth muscle cells (VSMCs) injury in rats.Methods The study was performed between March2022 and December 2022. VSMCs of cerebral basilar artery in rats were isolated and divided into blank group, H2O2 group (treatment with 500 μmol/L H2O2 for 6 h), H2O2+empty transfection group (treatment with 500 μmol/L H2O2 for 6 h, transfection with empty lipo-somes), H2O2+TRPC6 siRNA group (treatment with 500 μmol/L H2O2 for 6 h, transfection with TRPC6 siRNA) and H2O2 + pcD-NA3.1TRPC6 group (treatment with 500 μmol/L H2O2 for 6 h, transfection with pcDNA3.1TRPC6) according to random number tablemethod. The activity of cells proliferation was detected by human cholecystokinin/octapeptide (CCK)-8, cells apoptosis was detected by flow cytometry, levels of interleukin (IL)-6, IL-1β, monocyte chemotactic protein 1 (MCP-1), superoxide dismutase (SOD) and malondi-aldehyde (MDA) were detected by enzyme-linked immunosorbent assay, levels of reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe.Results Compared with H2O2 group, D(λ)450 nm was significantly increased in H2O2+TRPC6 siRNA group, while apoptosis rate was significantly decreased (P<0.05). Compared with H2O2 group, D(λ)450 nm was significantly decreased in H2O2+ pcDNA3.1 TRPC6 group, while apoptosis rate was significantly increased (P<0.05). In H2O2 group, H2O2+TRPC6 siRNA group and H2O2+pcDNA3.1 TRPC6 group, IL-6 levels were (159.24±21.65) ng/L, (52.95±12.43) ng/L and (257.48±23.44) ng/L, IL-1β levels were (126.09±19.85) ng/L, (41.67±9.06) ng/L and (248.77±23.32) ng/L, MCP-1 levels were (89.25±11.73) ng/L, (27.73±5.18) ng/L and(187.61±15.42) ng/L, SOD activities were (53.40±8.92) U/mL, (172.33±14.52) U/mL and (23.75±5.63) U/mL, malonaldehyde (MDA)levels were (91.35±10.41) nmol/mL, (8.29±2.17) nmol/mL and (169.37±14.37) nmol/mL, and ROS levels were 2.68±0.27, 0.76±0.18and 5.89±0.34. Compared with H2O2 group, IL-6, IL-1β, MCP-1, MDA and ROS were significantly decreased, while SOD activity was significantly increased in H2O2 +TRPC6 siRNA group (P<0.05), and all the above indexes were contrary in H2O2+pcDNA3.1 TRPC6 group.Conclusion The low/over expression of TRPC6 gene can promote/inhibit proliferative activity of H2O2 -induced VSMCs and in-hibit/promote cells apoptosis, which may be related to regulating inflammatory response and oxidative stress injury. |
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