云南泡核桃壳提取物体外抗肝纤维化活性研究

摘要

目的:研究云南泡核桃壳提取物(HTK-D、HTK-F、HTK-DEF)体外抗肝纤维化作用.方法:采用MTT法,观察云南泡核桃壳提取物对HSC增殖的影响及对L-02细胞的毒性作用.ELISA法检测细胞培养上清中TGF-β1、ColⅠ、ColⅢ的含量.结果:48、72 h时,HTK-D、HTK-DEF在15.63～1 000.00 μg/mL抑制HSC增殖(P＜0.05),HTK-F对HSC增殖抑制的浓度范围为62.5～1 000.00 μg/mL(P＜0.05).HTK-D、HTK-F、HTK-DEF分别与L-02细胞作用48 h后,其CC50分别为＞1 000.00、354.73、457.09 μg/mL.HTK-D、HTK-F、HTK-DEF在62.50～1 000.00 μg/mL时均能降低TGF-β1的生成(P＜0.05),且呈浓度依赖性;HTK-D、HTK-F、HTK-DEF分别在各浓度梯度下均能减少Col Ⅰ、ColⅢ的生成(P＜0.05).结论:云南泡核桃壳提取物具有体外抗肝纤维化作用,其作用机制可能与抑制肝星状细胞增殖,减少TGF-β1、Col Ⅰ、ColⅢ的生成有关.%Objective:To study the anti-hepatic fibrosis effects of extracts from walnut shell of Juglans sigillata D.in vitro(HTK-D,HTK-F,HTK-DEF).Methods:The MTT assay was used to observe the effects of the extracts from walnut shell of Juglans sigillata D.on hepatic stellate cells'(HSC)propagation and their cytotoxicity on normal human liver cell(L-02).ELISA method was used to detect the contents of TGF-β1,collagenⅠand collagenⅢin the supernatant of HSC.Results:Both HTK-D and HTK-DEF(the concentration from 15.63 μg/mL to 1 000.00 μg/mL)had inhibiting effects on HSC after 48 hours and 72 hours(P＜0.05).HTK-F(62.50 μg/mL-1 000.00 μg/mL)could inhibit the proliferation of HSC for 48,72 h(P＜0.05).The values of CC 50were＞1 000.00 μg/mL,354.73 μg/mL,457.09 μg/mL accordingly after L-02 was cultured with HTK-D,HTK-F and HTK-DEF respectively for 48 h.HTK-D,HTK-F and HTK-DEF could reduce the production of TGF-β1 in supernatant for 48 h at concentration between 62.50 μg/mL and 1 000.00 μg/mL in a concentration dependent manner(P＜0.05).HTK-D,HTK-F and HTK-DEF could decrease the production of collagenⅠand collagenⅢin supernatant at different concentration gradient(P＜0.05).Conclusion:Extracts from walnut shell of Juglans sigillata D.in vitro have definite effects on anti-hepatic fibrosis.The mechanism may be related to inhibiting HSC propagation and reducing the production of TGF-β1,collagenⅠand collagenⅢ.