醋制降低甘遂对人正常肝细胞LO2毒性研究

Study on detoxication of Kansui Radix on normal liver cells LO2 after stir-baking with vinegar

目的:比较甘遂醋制前后对人正常肝细胞LO2的毒性差异,初步探讨甘遂醋制减毒机制。方法:以人正常肝细胞LO2细胞为研究对象,采用MTT法检测甘遂醋制前后对LO2细胞活性的影响,采用倒置显微镜观察细胞形态;并测定细胞培养上清液和裂解上清液中谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、Na+-K+-ATP酶,Ca2+-Mg2+-ATP酶等酶的活力和谷胱甘肽(GSH)、丙二醛(MDA)的含量。结果:与阴性对照组比较,生甘遂可明显降低LO2细胞的活性(P<0.01)与形态,并显著提高LO2细胞培养上清液中ALT,AST,LDH活力(P<0.01)、显著降低SOD活力和GSH含量(P<0.01),显著增加MDA含量(P<0.01),显著降低LO2细胞裂解上清液中Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活力(P<0.01);醋制后,与生甘遂各剂量组比较,醋甘遂可显著降低生甘遂对LO2细胞的增殖抑制作用(P<0.01)和形态变差的趋势,可显著降低LO2细胞培养上清液中ALT,AST,LDH活力(P<0.01)、显著增加SOD活力和GSH含量(P<0.01)、显著降低MDA含量(P<0.01)、显著提高LO2细胞裂解上清液中Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活力(P<0.01),且呈一定的剂量相关性。结论:醋制可降低甘遂肝毒性,其可能机制为通过降低甘遂对肝细胞膜通透性的影响及减轻氧化损伤而实现。

Objective： To compare the toxicity on normal liver cells LO2 before and after Kansui Radix stir-baked with vine- gar, and make a preliminary study on the mechanism of detoxication of Kansui Radix stir-baked with vinegar. Method： The MTT method was adopted to detect the cell activity, with normal liver cells LO2 as the study object. The morphology of cells were observed, and the level or content of AST, ALT, LDH, SOD, Na^＋ -K^＋ -ATPase, Ca^2＋ -Mg^2＋ -ATPase, GSH and MDA were determined in cell culture supernatant and splitting supernatant. Result： Compared with the control group, Kansui can obviously inhibit the cell activity （ P 〈 0.01 ） and morphology, and increase the levels of ALT, AST, and LDH （ P 〈 0. 01 ） in the supematant fluid of cell incubation, and decrease the level of SOD and the content of GSH （P 〈0. 01 ）. Besides, it significantly increased the content of MDA （P 〈0. 01 ） and significantly decreased the level of Na^＋ -K^＋ -ATPase and Ca^2＋ -Mg^2＋ -ATPase （P 〈 0.01 ） in the supernatant fluid of cell dissocia- tion. Compared with Kansui group of various doses, Kansui Radix stir-baked with vinegar can significantly decrease the cell prolifera- tion inhibition and the trend of morphological variation, and obviously decrease the levels of ALT, AST, and LDH （P 〈 0. 01 ） in the supematant fluid of cell incubation, and significantly increase the level of SOD and the content of GSH （P 〈 0. 01 ）, and significantly decrease the content of MDA （P 〈 0. 01）. Additionally, it significantly increased the level of Na^＋-K^＋-ATPase and Ca^2＋-Mg^2＋- ATPase （P 〈 0. 01 ） in the supematant fluid of cell dissociation, and showed a certain dose-effect relationship. Conclusion： Stir-bak- ing with rice vinegar can release the hepatotoxicity of Kansui Radix. Its possible mechanism was that Kansui Radix stir-baked with vine- gar can decrease the influence of Kansui Radix on the permeability of liver cells LO2 membrane and oxidative damage, in order to pro- vide basis for further exploration of the detoxication mechanism of Kansui Radix stir-baked with vinegar.