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本研究探讨了丹参酮IIA对脂多糖诱导大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMCs)炎症反应、氧化应激及其损伤的影响。采用原代培养大鼠腹膜间皮细胞(RPMCs),分为正常对照组、5 mg/L LPS作用RPMC 24 h组、5 mg/L LPS分别与40、80和160μmol/L丹参酮IIA共同作用24 h组。MTT测定各组RPMCs增值率。ELISA法检测细胞培养液中IL-1β、IL-6和TNF-α表达。流式细胞仪检测活性氧(ROS)水平,试剂盒检测丙二醛(MDA)和超氧化物歧化酶(SOD)表达。RT-PCR法检测各组FN、COL I、Bcl-2和Bax mRNA的表达。研究发现丹参酮IIA+LPS组的RPMCs的增殖率明显高于LPS组(P<0.05)。丹参酮IIA可降低LPS刺激下RPMCs中IL-1β、IL-6、TNF-α、ROS和MDA的表达,同时FN、COL I、Bax mRNA表达也明显下降。但SOD水平和Bcl-2 mRNA表达明显升高,与LPS组相比。实验结果显示,丹参酮IIA具有抑制LPS所致的氧化应激及炎性反应,减少细胞凋亡及抑制纤维化的作用,进而起到对RPMCs的保护作用。
This study investigated the effects of tanshinone IIA on the inflammatory response, oxidative stress and its damage induced by lipopolysaccharide in rat peritoneal mesothelial cells (RPMCs). Primary cultured rat peritoneal mesothelial cells (RPMCs) were divided into normal control group, RPMCs treated with 5 mg / L LPS for 24 h and 5 mg / L LPS with 40, 80 and 160 μmol / L tanshinone IIA respectively h group. MTT assay RPMCs in each group value-added rate. ELISA was used to detect the expression of IL-1β, IL-6 and TNF-α in cell culture medium. The level of reactive oxygen species (ROS) was detected by flow cytometry. The expression of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by kit. The mRNA expression of FN, COL I, Bcl-2 and Bax in each group was detected by RT-PCR. The results showed that the proliferation rate of RPMCs in tanshinone IIA + LPS group was significantly higher than that in LPS group (P <0.05). Tanshinone IIA could reduce the expression of IL-1β, IL-6, TNF-α, ROS and MDA in RPMCs stimulated by LPS, meanwhile, the expression of FN, COL I and Bax mRNA also decreased obviously. However, the level of SOD and the expression of Bcl-2 mRNA were significantly increased, compared with LPS group. The results showed that tanshinone IIA can inhibit the oxidative stress and inflammatory response induced by LPS, reduce apoptosis and fibrosis, and then play a protective role on RPMCs.