1. 上海中医药大学附属龙华医院消化内镜科,上海,200032
2. 上海中医药大学附属龙华医院脾胃病科,上海,200032
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史琲, 黄傲霜, 胡鸿毅, 等. 清肠栓对LPS诱导Caco-2细胞损伤的保护作用[J]. 上海中医药杂志, 2019,53(6):69-74.
SHI Bei, HUANG Aoshuang, HU Hongyi, et al. Protective effects of Qingchang Suppository on LPS-induced Caco-2 cell injury[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):69-74.
史琲, 黄傲霜, 胡鸿毅, 等. 清肠栓对LPS诱导Caco-2细胞损伤的保护作用[J]. 上海中医药杂志, 2019,53(6):69-74. DOI: 10.16305/j.1007-1334.2019.06.018.
SHI Bei, HUANG Aoshuang, HU Hongyi, et al. Protective effects of Qingchang Suppository on LPS-induced Caco-2 cell injury[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):69-74. DOI: 10.16305/j.1007-1334.2019.06.018.
目的:探讨清肠栓(QCS)含药血清对脂多糖(LPS)诱导Caco-2细胞损伤模型的保护作用。 方法:收集对数生长期细胞,分为5个组,即正常对照组、LPS组,以及QCS低剂量组(2.5%含药血清)、中剂量组(5%含药血清)、高剂量组(10%含药血清)。采用MTT法检测细胞活力;ELISA法测定培养细胞上清液IL-8、IL-17和PGE,2,含量,以及MDA、GSH和SOD水平;划痕实验检测细胞体外迁移能力;Western blot技术检测RhoA、Rac1蛋白的表达。 结果:QCS能拮抗LPS诱导的Caco-2细胞活性下降;与LPS组比较,QCS可显著减少LPS诱导的IL-8、IL-17和 PGE,2,分泌(P,<,0.05),并减少LPS诱导的MDA释放,增加GSH和SOD的分泌(P,<,0.01);划痕实验48 h时QCS组的划痕宽度明显变小,细胞迁移能力增强(P,<,0.001),划痕的修复加快,且呈浓度依赖性改变;Western blot结果显示,QCS能有效逆转LPS所致RhoA、Rac1蛋白表达的抑制(P,<,0.05)。 结论:QCS含药血清对LPS诱导的Caco-2细胞损伤有保护作用,可通过抑制炎症因子、抗氧化应激和促进结肠上皮细胞迁移来提高Caco-2的生存率。
Objective:To investigate the protective effects of Qingchang Suppository (QCS) medicated serum on lipopolysaccharide (LPS)-induced Caco-2 cell injury model. MethodsCaco-2 cells in logarithmic growth phase were collected and divided into five groups: normal control group, LPS group, low-dose QCS group (2.5% QCS medicated serum), medium-dose QCS group (5% QCS medicated serum) and high-dose QCS group (10% QCS medicated serum). MTT assay was used to detect cell viability; ELISA was used to determine the content of IL-8, IL-17 and PGE,2, and the levels of MDA, GSH and SOD in the supernatant; scratch assay was employed to test cell migration ability in vitro; Western blot was used to detect the expression of RhoA and Rac1 proteins. Results:QCS antagonized the decrease in Caco-2 cell viability induced by LPS. Compared with the LPS group, QCS significantly reduced the secretion of IL-8, IL-17 and PGE,2, induced by LPS (P,<,0.05), lowered the release of MDA induced by LPS and increased the secretion of GSH and SOD (P,<,0.01); the scratch width in the QCS group decreased significantly and the cell migration ability increased at 48 h (P,<,0.001), which speeded up the repair of scratches in a concentration-dependent manner; Western blot results showed that QCS could effectively reverse LPS-induced inhibition of RhoA and Rac1 protein expression (P,<,0.05). Conclusion:Qingchang Suppository medicated serum protects Caco-2 cells from LPS-induced injury and increases cell survival rate by inhibiting inflammatory factors and antioxidant stress and promoting migration of colonic epithelial cells.
清肠栓Caco-2细胞脂多糖RhoARac1细胞培养
Qingchang SuppositoryCaco-2 cellslipopolysaccharideRhoARac1cell culture
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