
浏览全部资源
扫码关注微信
纸质出版:2013
移动端阅览
曹敏,黄家君.小檗碱减轻炎症反应时肠上皮屏障功能损害的实验研究[J].陆军军医大学学报(原第三军医大学学报),2013,35(05):447-450.
Cao Min,Huang Jiajun.Attenuation of intestinal epithelial barrier disruption by berberin during inflammation in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(05):447-450.
目的 研究小檗碱(berberine,BBR)在炎症反应时对肠上皮屏障功能的保护作用。 方法 建立人肠上皮细胞株Caco-2单层培养模型,将单层细胞分为对照组、BBR组、TNF-α+IFN-γ组、TNF-α+IFN-γ+BBR组。用电阻仪测定跨单层肠上皮细胞电阻(TER),测定时相点为处理后0、6、12、24、36 h和48 h;免疫荧光法和蛋白质印迹法检测紧密连接蛋白Occludin的分布及表达变化;蛋白质印迹法检测肌球蛋白轻链(MLC)和磷酸化肌球蛋白轻链(pMLC)的蛋白表达。 结果 对照组与BBR组TER均无明显变化;TNF-α+IFN-γ组TER逐渐下降,并于12、24、36 h及48 h显著低于0 h(P0.05);TNF-α+IFN-γ+BBR组也呈逐渐下降趋势,但降幅小于TNF-α+IFN-γ组,并于12、24、36 h及48 h显著高于TNF-α+IFN-γ组(P0.05)。各组紧密连接蛋白Occludin表达无明显变化(P0.05);但TNF-α+IFN-γ组Occludin形态有明显变化,并发生重分布,而TNF-α+IFN-γ+BBR组Occludin形态变化则较TNF-α+IFN-γ组明显减轻。TNF-α+IFN-γ组pMLC表达显著高于对照组或BBR组(P0.05),而TNF-α+IFN-γ+BBR组pMLC表达则明显低于TNF-α+IFN-γ组(P0.05)。 结论 小檗碱通过降低MLC磷酸化水平,改善紧密连接蛋白定位分布变化,从而减轻炎症反应时肠上皮屏障功能损害。
Objective To investigate the protective effect of berberine (BBR) on intestinal epithelial barrier function during inflammation and the underlying mechanism. Methods The intestinal epithelial cell line Caco-2 monolayers were divided into a control group
a BBR group
a TNF-α+IFN-γ group and a TNF-α+IFN-γ+BBR group. Both of TNF-α and IFN-γ were used at the final concentration of 10 ng/mL
and BBR was used at 100 μmol/L. The transepithelial electrical resistance (TER) was measured with a Volt-Ohm Meter at 0
6
12
24
36 and 48 h after treatment. Both Western blot analysis and immunofluorescence assay were used to analyze the tight junction protein occludin. The protein expression of myosin light chain (MLC) and phosphorylated MLC (pMLC) was analyzed by Western blot analysis. Results The TER had no significant change in the control group and BBR group after treatment. In the TNF-α + IFN-γ group
TER decreased gradually after treatment
and was significantly lower at 12
24
36 and 48 h than that before treatment (P0.05). In the TNF-α + IFN-γ + BBR group
TER also decreased gradually after treatment
but was significantly higher than that in the TNF-α + IFN-γ group at 12
24
36 and 48 h after treatment (P0.05). The protein expression of occludin in each group did not change significantly after treatment (P0.05). However
the remarkable morphological relocalization with expanded intracellular pools of occludin was observed in the TNF-α + IFN-γ group
which was attenuated obviously in the TNF-α + IFN-γ + BBR group. The pMLC protein expression in the TNF-α + IFN-γ group was significantly higher than that in other 3 groups (P0.05). Conclusion BBR alleviates intestinal epithelial barrier disruption during inflammation by reducing MLC phosphorylation level and attenuating the morphological relocalization of tight junction protein occludin.
0
浏览量
0
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621