参苓白术散含药血清抗脂多糖(LPS)诱导的肠隐窝上皮细胞(IEC-6)通透性变化机制的研究
| 作者: |
1黄玉海,
2刘玉晖,
3廖旺娣,
3游宇
1 江西萍乡市第二人民医院普外科，江西 萍乡 337000 2 江西中医药大学药学院，南昌 330004 3 南昌大学第一附属医院消化科，南昌 330006 |
| 通讯: |
游宇
Email: youyu1979@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2015.12.012 |
| 基金: | 国家自然科学基金项目, 81160540/H2814 |
摘要
目的：探讨参苓白术散抗脂多糖(lipopolysaccharide，LPS)诱导肠上皮隐窝细胞(intestinal cryptcells，IEC-6)通透性变化的机制研究。方法：将IEC-6细胞分为空白组、LPS模型组、LPS+参苓白术散含药血清低、中、高剂量组、10%FBS。对以下指标进行检测：IEC-6细胞凋亡、细胞内钙离子浓度、IEC-6电阻值变化、检测各组中磷酸化Rho激酶(ROCKⅡ)、肌球蛋白轻链激酶(MLCK)以及凋亡因子Caspase-3表达。结果：LPS显著引起IEC-6细胞凋亡而参岺白术散的含药血清明显抑制细胞凋亡。与空白组比较，模型组IEC-6细胞跨膜电阻的TEER值显著下降(P<0.01)；与LPS模型组比较，参苓白术散含药血清低、中、高剂量组IEC-6细胞跨膜电阻的TEER值显著升高，实验结果具有统计学意义(P<0.01)。LPS模型组的钙离子浓度明显高于空白组(P<0.01)；不同浓度的参苓白术散含药血清组的钙离子浓度明显低于LPS模型组(P<0.01)。与空白组比较，模型组中磷酸化ROCKⅡ、MLCK蛋白表达明显升高(P<0.05)；与模型组比较，不同剂量参苓白术散含药血清组中磷酸化ROCKⅡ、MLCK蛋白表达明显降低(P<0.05)。结论：参苓白术散含药血清对LPS诱导的IEC-6细胞损伤具有明显地抑制作用，与抑制细胞凋亡及IEC-6细胞通透性的改善有关。
关键词：
肠上皮隐窝细胞
参苓白术散
脂多糖
细胞通透性
Rho激酶
The study of mechanism the serum of SLBZS improve perpeability change on IEC-6 induced by LPS
CorrespondingAuthor: YOU Yu Email: youyu1979@126.com
DOI: 10.3978/j.issn.2095-6959.2015.12.012
Abstract
Objective: To investigate the serum of SLBZS treat permeability change induced by lipopolysaccharide (LPS) on intestinal crypt cells (IEC-6) study of the mechanism. Methods: IEC-6 cells were divided into control group, model group, Shenling Baizhu powder of low (4%), medium (8%) and high (16%) dose group with LPS, 10%FBS. Building inflammation model of IEC-6 cells with 200 μg/mL lipopolysaccharide. The Annexin V-FITC/PI Hoechst staining were used to detect the LPS on IEC-6 cell apoptosis. The Flou 3-AM method of timeresolved was used to detect the intracellular calcium ion concentration in each group. Test the IEC-6 cell TEER, Western blot detected the Phosphorylated ROCKII, MLCK, and the expression of apoptosis factor Caspase-3. Results: LPS significantly induced IEC-6 apoptosis and was reduced by SLBZS serum. The calcium ion concentration of the model group was significantly higher than the control group (P<0.01), Shenling Baizhu powder containing serum group and different concentration of calcium ion concentration was significantly lower than that of model group (P<0.01). The TEER value of IEC-6 cells, compared with blank group, model group transepithelial electrical resistance of IEC–6 decreased significantly (P<0.01); compared with model group, and SLBZS serum levels of low, medium and high dose group of IEC-6 across cell membrane resistance TEER a significant rise in value (P<0.01). The model group compared with the control group, the expression of Pho-ROCK II, MLCK protein increased significantly (P<0.05); while different dose medicated serum group could decrease the expression of Pho-ROCKII, MLCK protein decreased (P<0.05). Conclusion: SLBZS drugcontaining serum can significant inhibit IEC-6 cell damage induced by LPS, it may be related to inhibit cell apoptosis and the improvement of the IEC-6 cell permeability.