不同水分处理垂盆草对CCl_4致HepG2细胞损伤的保护作用
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摘要
目前探讨栽培措施对垂盆草药材药理活性影响的研究尚不多见。为了探索不同水分处理垂盆草的保肝效果差异,以不同水分梯度[田间持水量的15%~20%(S1),35%~40%(S2),55%~60%(S3),75%~80%(S4),95%~100%(S5)]处理60 d后垂盆草的水提物为研究材料,探索其对四氯化碳(CCl_4)诱导HepG2细胞氧化损伤的预防效果。MTT法检测HepG2细胞存活率,PI单染法检测细胞周期,Annexin-V FITC/PI双染法检测细胞凋亡,比色法测定细胞上清液中转氨酶及抗氧化指标。结果表明,与模型组相比,垂盆草水提物能显著减轻CCl_4导致的HepG2细胞损伤,提高HepG2细胞存活率。5个垂盆草组处理的保肝作用有显著差异,以75%~80%田间持水量(S4)处理垂盆草水提液的HepG2细胞存活率、SOD和GSH水平最高,细胞凋亡率,ALT,AST,MDA水平最低;而95%~100%田间持水量(S5)处理的HepG2细胞存活率最低,凋亡率和转氨酶活性最高,SOD,GSH,GSH-Px水平最低。综上所述,不同水分处理对垂盆草的保肝活性有较大影响,75%~80%田间持水量条件下处理60 d的垂盆草保肝作用最佳。
At present,there were few studies about the effects of cultivation measures on the quality and pharmacological activity of medicinal plants. To explore the hepetoprotective activity of Sedum sarmentosum aqueous extracts after different water treatments,S. sarmentosum were planted under five water treatments for 60 days,namely 15%-20% FC(field capacity,S1),35%-40% FC(S2),55%-60% FC(S3),75%-80% FC(S4),and 95%-100% FC(S5) and CCl_4 drug-induced liver injury model in vitro was used. Cell viability,cell cycle,and cell apoptosis were individually detected by MTT,PI single staining,and Annexin-V FITC/PI double staining assays. Additionally,ALT,AST and antioxidant index in supernatant were determined by colorimetry. The results showed that,compared with the model group,S. sarmentosum aqueous extract could significantly improve the HepG2 cell viability.Among the five S. sarmentosum groups,the cell viability of S4(75%-80% FC) treatment was the highest,and the cell apoptosis was the least. Meanwhile,the level of ALT,AST,and MDA in S4 was the lowest,but it achieved the highest level of SOD and GSH.Taken together,different water treatments had great influence on hepatoprotective effect of S. sarmentosum,and the soil moisture of the75%-80% FC is beneficial to the hepetoprotective activity of S. sarmentosum.
At present,there were few studies about the effects of cultivation measures on the quality and pharmacological activity of medicinal plants. To explore the hepetoprotective activity of Sedum sarmentosum aqueous extracts after different water treatments,S. sarmentosum were planted under five water treatments for 60 days,namely 15%-20% FC(field capacity,S1),35%-40% FC(S2),55%-60% FC(S3),75%-80% FC(S4),and 95%-100% FC(S5) and CCl_4 drug-induced liver injury model in vitro was used. Cell viability,cell cycle,and cell apoptosis were individually detected by MTT,PI single staining,and Annexin-V FITC/PI double staining assays. Additionally,ALT,AST and antioxidant index in supernatant were determined by colorimetry. The results showed that,compared with the model group,S. sarmentosum aqueous extract could significantly improve the HepG2 cell viability.Among the five S. sarmentosum groups,the cell viability of S4(75%-80% FC) treatment was the highest,and the cell apoptosis was the least. Meanwhile,the level of ALT,AST,and MDA in S4 was the lowest,but it achieved the highest level of SOD and GSH.Taken together,different water treatments had great influence on hepatoprotective effect of S. sarmentosum,and the soil moisture of the75%-80% FC is beneficial to the hepetoprotective activity of S. sarmentosum.
引文
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[20]吴琼,刘香香,陆定艳,等.荭草花提取物对H2O2诱导的HUVEC细胞氧化损伤的保护作用研究[J].中国中药杂志,2018,43(5):1008.
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[22]闫冰,丁安伟,张丽.二至丸提取物对小鼠四氯化碳急性肝损伤的保护作用[J].中国中药杂志,2010,35(22):3080.
[23]潘金火,潘萍.垂盆草总黄酮中8种单体成分对肝细胞的保护作用[J].中国医院药学杂志,2010,30(19):1621.
[24]张石蕾,由淑萍,刘涛,等.睡莲花总黄酮对四氯化碳致小鼠急性肝损伤的防治作用[J].天然产物研究与开发,2016,28(12):2017.
[25]Fornaciari S,Milano F,Mussi F,et al. Assessment of antioxidant and antiproliferative properties of spinach plants grown under low oxygen availability[J]. J Sci Food Agric,2015,95(3):490.
[26]Chrysargyris A,Xylia P,Botsaris G,et al. Antioxidant and antibacterial activities, mineral and essential oil composition of spearmint(Mentha spicata L.)affected by the potassium levels[J]. Ind Crop Prod,2017,103:202.
[27]Kim J M,Yoon K Y. Comparison of polyphenol contents,antioxidant,and anti-inflammatory activities of wild and cultivated Lactuca indica[J]. Hort Environ Biotechnol,2014,55(3):248.
[2]全吉淑,王玉娇,尹基峰,等.草苁蓉多糖对HepG2细胞氧化应激的抑制作用[J].食品研究与开发,2016,37(11):6.
[3]Niki E. Assessment of antioxidant capacity in vitro and in vivo[J]. Free Radical Bio Med,2010,49(4):503.
[4]刘红亮,胡磊,王靖凯,等.槲皮素对H2O2损伤PC12细胞的保护效果与机制[J].中国药理学通报,2014,30(3):373.
[5]中国科学院中国植物志编辑委员会.中国植物志.第34卷.第1分册[M].北京:科学出版社,1984:146.
[6]中国药典.一部[S]. 2015.
[7]张洪超,兰天,张晓辉.垂盆草化学成分与药理作用研究进展[J].中成药,2005,27(10):1201.
[8]鲁合军.垂盆草水煎液对4种肝损伤模型小鼠的保肝作用[J].中医学报,2017,32(3):409.
[9]孙卫东,胡海涛,钱山青,等.垂盆草提取液对小鼠免疫和抗氧化功能的研究[J].中外医疗,2008,27(31):19.
[10]杨金凤,郭巧生,朱再标,等.不同水分梯度下垂盆草生长发育、品质及抗氧化活性关系研究[J].中草药,2018,49(14):3382.
[11]伍振辉,孟娴,丁永芳,等.白首乌C-21甾体总苷对人肝癌HepG2细胞增殖、侵袭与凋亡的影响[J].中国中药杂志,2018,43(9):1915.
[12]González L T,Minsky N W,Espinosa L E M,et al. In vitro assessment of hepatoprotective agents against damage induced by acetaminophen and CCl4[J]. BMC Complem Altern M,2017,17(1):39.
[13]Krithika R,Mohankumar R,Verma R J,et al. Isolation,characterization and antioxidative effect of phyllanthin against CCl4-induced toxicity in HepG2 cell line[J]. Chem Biol Interact,2009,181(3):351.
[14]卢春远,林兴,黄权芳,等.山芝麻酸甲酯对CCl4致大鼠慢性肝损伤的作用及其机制的影响[J].中国实验方剂学杂志,2017,23(7):141.
[15]王凯,眭丹娟,王长锁,等. 5种石斛对四氯化碳致小鼠急性肝损伤的保护作用[J].中国中药杂志,2017,42(10):1945.
[16]刘三海,李蕊,乔艳雪,等.垂盆草水煎液对不同小鼠急性肝损伤模型的疗效作用[J].中国肝脏病杂志:电子版,2015,7(1):37.
[17]黄静,李胜立,赵宏伟,等.霍山石斛多糖对四氯化碳致急性肝损伤小鼠的保护作用[J].中国中药杂志,2013,38(4):528.
[18]苏华健,陈雨晴,李晓萱,等.富硒乳酸菌对四氯化碳诱导的大鼠肝损伤的保护作用[J].南京农业大学学报,2016,39(4):661.
[19]王玉荣,杨康,崔伟业,等.绞股蓝黄酮对过氧化氢损伤A549细胞的作用[J].中国中药杂志,2018,43(5):1014.
[20]吴琼,刘香香,陆定艳,等.荭草花提取物对H2O2诱导的HUVEC细胞氧化损伤的保护作用研究[J].中国中药杂志,2018,43(5):1008.
[21]Chen Q,Zhan Q,Li Y,et al. Schisandra lignan extract protects against carbon tetrachloride-induced liver injury in mice by inhibiting oxidative stress and regulating the NF-κB and JNK signaling pathways[J]. Evid-Based Compl Alt,2017,2017:1.
[22]闫冰,丁安伟,张丽.二至丸提取物对小鼠四氯化碳急性肝损伤的保护作用[J].中国中药杂志,2010,35(22):3080.
[23]潘金火,潘萍.垂盆草总黄酮中8种单体成分对肝细胞的保护作用[J].中国医院药学杂志,2010,30(19):1621.
[24]张石蕾,由淑萍,刘涛,等.睡莲花总黄酮对四氯化碳致小鼠急性肝损伤的防治作用[J].天然产物研究与开发,2016,28(12):2017.
[25]Fornaciari S,Milano F,Mussi F,et al. Assessment of antioxidant and antiproliferative properties of spinach plants grown under low oxygen availability[J]. J Sci Food Agric,2015,95(3):490.
[26]Chrysargyris A,Xylia P,Botsaris G,et al. Antioxidant and antibacterial activities, mineral and essential oil composition of spearmint(Mentha spicata L.)affected by the potassium levels[J]. Ind Crop Prod,2017,103:202.
[27]Kim J M,Yoon K Y. Comparison of polyphenol contents,antioxidant,and anti-inflammatory activities of wild and cultivated Lactuca indica[J]. Hort Environ Biotechnol,2014,55(3):248.