苦杏仁苷对血管紧张素Ⅱ刺激HSC-T6细胞Ca^(2+)效应的抑制作用  

Inhibitory Effect of Amygdalin on Ca^(2+) Effect of Angiotensin Ⅱ-stimulated Hepatic Stellate Cell- T6

作　　者：骆欢欢[1] 赵昉[1] 张奉学[1] 刘妮[1] 王婷[1] 

机构地区：[1]广州中医药大学,广东广州510405

出　　处：《广州中医药大学学报》2016年第2期216-219,共4页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广东省自然科学基金资助项目(编号:S2012100008917)

摘　　要：【目的】观察苦杏仁苷对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)刺激下HSC-T6细胞Ca2+效应的抑制作用。【方法】将HSC-T6细胞系分为以下5组:正常组、AngⅡ组、苦杏仁苷高剂量(10-3 mol/L)组、苦杏仁苷中剂量(10-4 mol/L)组、苦杏仁苷低剂量(10-5 mol/L)组。正常组采用含体积分数2%胎牛血清的DMEM培养,其余各组在DMEM培养液的基础上,AngⅡ组加入10-7 mol/L AngⅡ溶液;高、中、低剂量苦杏仁苷组均加入10-7 mol/L AngⅡ溶液,在此基础上各组分别加入不同浓度的苦杏仁苷。钙荧光探针Fluo-3/AM负载后置于LSCM SP2激光共聚焦显微镜(LSCM)下检测Ca2+。【结果】LSCM下Ca2+荧光成像的HSC-T6细胞仍可部分表现倒置显微镜下的形态学特征,各组细胞均染色良好,且各细胞间染色强度基本一致。AngⅡ组Ca2+荧光强度非常明显,清晰可见;正常组强度也较明显,稍次于AngⅡ组;苦杏仁苷高剂量组强度也较强,稍次于AngⅡ组和正常组,但区别不甚明显;苦杏仁苷中、低剂量组强度明显减弱,并可见大量Ca2+噬斑出现,其中中剂量组图像噬斑更为清晰。AngⅡ组荧光强度较正常组稍强,但差异无统计学意义(P>0.05)。苦杏仁苷高剂量荧光强度稍低于AngⅡ组与正常组,但差异均无统计学意义(P>0.05);苦杏仁苷中、低剂量组荧光强度显著低于AngⅡ组和正常组,差异均有统计学意义(P<0.05)。【结论】苦杏仁苷可抑制HSC内Ca2+升高,从而抑制HSC的活化与增殖,产生抗肝纤维化作用。Objective To observe the inhibitory effect of amygdalin on Ca^2＋ effect of angiotensin（Ang） Ⅱ-stimulated hepatic stellate cell-T6（HSC-T6）. Methods HSC-T6 cell line was divided into 5 groups,namely normal group,AngⅡgroup,and high-,middle- and low-dose amygdalin groups（amygdalin at 10-3,10-4,10-5mol/L,respectively）. The normal group was treated with 2% of DMEM. Ang Ⅱ group and the three amygdalin groups were treated with 2% of DMEM and 10-7mol/L AngⅡ,and additionally the amygdalin groups were given the corresponding concentrations of amygdalin. Ca2 ＋was detected under the laser scanning confocal microscope（LSCM）SP2 after calcium fluorescence probe Flou-3/AM was loaded. Results Under the LSCM,HSC-T6 cells with Ca2 ＋fluorescence imaging showed the partial morphological characteristics like those under the ordinary inverted microscope： the staining of the cell was well, and the intercellular staining intensity was even. Ca2 ＋fluorescence imaging in Ang II group was clear and the fluorescence intensity was the highest, and then came the normal group and high-dose amygdalin group, but the differences were not significant. The intensity in middle- and low- dose amygdalin groups was obviously decreased and large amount of plaque Ca2 ＋was seen,and Ca2 ＋fluorescence imaging in middle-dose amygdaline group was more clear. The fluorescence intensity of Ang II group was higher than that of the normal group, and the intensity of high-dose amygdaline was lower than that of Ang II group and the normal group, but the difference was all insignificant（P〈0.05）. The fluorescence intensity of middle- and low-dose amygdaline groups was lower than that of Ang II group and the normal group,and the difference was significant（P〈0.05）. Conclusion Amygdaline has inhibitory effect on the increase of Ca2 ＋in HSC, thereby can inhibit the activation and proliferation of HSC, and conteract hepatic fibrosis.

关 键 词：苦杏仁苷/药理学 肝纤维化/中药疗法 肝星状细胞/病理学 细胞培养 钙离子 

分 类 号：R285.5[医药卫生—中药学]