Cao Yu-Dan, Yan Xiao-Jing, Zhang Li, Ding An-Wei
Zhongguo Zhong Yao Za Zhi. 2014 Mar;39(6):1069-74.
To compare the difference of Euphorbia Pekinensis Radix before and after being processed with vinegar in the toxicity on rat small intestinal crypt epithelial cells IEC-6, and make a preliminary study on the mechanism of detoxication of Euphorbia Pekinensis Radix processed with vinegar.
With rat small intestinal crypt epithelial cells IEC-6 as the study object, the MTT method was adopted to detect the effect of Euphorbia Pekinensis Radix before and after being processed with vinegar on IEC-6 cell activity. The morphology of cells were observed by the inverted microscope. The down-regulated mitochondrial apoptosis pathway of enterocytes caused by the vinegar processing was analyzed by using the high content screening.
Compared with the negative control group, the proliferation inhibition experiment showed that Euphorbia Pekinensis Radix showed a relatively high intestinal cell toxicity (P < 0.01). The results of HCS analysis showed that Euphorbia Pekinensis Radix could significantly reduce the cell nucleus Hoechst fluorescence intensity and mitochondria membrane (P < 0.05, P < 0.01), and increase Annexin V-FITC and PI fluorescence intensity and membrane permeability (P < 0.01, P < 0.01, P < 0.01). After being processed with vinegar, compared with Euphorbia Pekinensis Radix groups with different doses, Euphorbia Pekinensis Radix processed with vinegar could significantly decrease the cell proliferation inhibition effect on enterocytes, increase the cell nuclear Hoechst fluorescence intensity and mitochondria membrane (P < 0.05, P < 0.05), and decrease Annexin V-FITC and PI fluorescence intensity and membrane permeability (P < 0.01, P < 0.01, P < 0.05), and showed a certain dose-effect relationship.
The vinegar processing can further reduce the toxicity of Euphorbia Pekinensis Radix on enterocytes. Its possible mechanism can decrease the effect of Euphorbia Pekinensis Radix on the permeability of IEC-6 cell membrane, so as to provide a basis for further explanation of the detoxication mechanism of Euphorbia Pekinensis Radix processed with vinegar.
比较醋制前后京大戟对大鼠小肠隐窝上皮细胞IEC-6毒性的差异,并对醋制京大戟的解毒机制进行初步研究。
以大鼠小肠隐窝上皮细胞IEC-6为研究对象,采用MTT法检测醋制前后京大戟对IEC-6细胞活性的影响。用倒置显微镜观察细胞形态。利用高内涵筛选分析醋制引起的肠上皮细胞线粒体凋亡途径下调情况。
增殖抑制实验显示,与阴性对照组相比,京大戟对肠细胞具有较高毒性(P<0.01)。高内涵筛选分析结果显示,京大戟可显著降低细胞核Hoechst荧光强度和线粒体膜电位(P<0.05,P<0.01),增加Annexin V-FITC和PI荧光强度及膜通透性(P<0.01,P<0.01,P<0.01)。醋制后,与不同剂量的京大戟组相比,醋制京大戟可显著降低对肠上皮细胞的增殖抑制作用,增加细胞核Hoechst荧光强度和线粒体膜电位(P<0.05,P<0.05),降低Annexin V-FITC和PI荧光强度及膜通透性(P<0.01,P<0.01,P<0.05),并呈现一定的剂量效应关系。
醋制可进一步降低京大戟对肠上皮细胞的毒性。其可能机制为降低京大戟对IEC-6细胞膜通透性的影响,从而为进一步阐释醋制京大戟的解毒机制提供依据。