Na~+/K~+ATPase介导甘草诱导低钾血症的研究
|
摘要
目的探讨高剂量甘草水提物导致小鼠低钾血症的分子机制。方法利用高剂量甘草水提物(7.5 g/kg)灌胃制备低钾血症小鼠模型,收集小鼠尿液和粪便,检测其K~+浓度;分析小鼠肠道组织的转录组数据,寻找与钾代谢相关基因的表达变化,并利用real-time PCR进行确认;针对显著改变的关键基因进行干预,确认其与甘草诱导低钾血症的关系。结果高剂量甘草水提物可造成小鼠低钾血症,转录组结果表明Na~+/K~+ATPase的表达水平发生显著变化,real-time PCR结果也确证了这一点,肠道病理切片显示肠道通透性发生改变;低钾血症小鼠的尿钾降低,而粪便中K~+浓度增多,表明甘草水提物诱导的低钾血症可能主要由肠道失钾造成;给予Na~+/K~+ATPase抑制剂处理后,尿液中K~+浓度快速升高,粪便中的K~+浓度降低;继续予以高剂量甘草水提物处理,在Na~+/K~+ATPase抑制剂药效消除后小鼠尿液和粪便中K~+浓度又恢复至造模状态。结论高剂量甘草水提物造成的低钾血症主要是由胃肠道失钾造成的,而这种低钾血症可以通过Na~+/K~+ATPase抑制剂特异性地进行逆转。
Objective To explore the molecular mechanism of hypokalemia induced by high dose of Glycyrrhiza uralensis decoction in mice.Methods Hypokalemia mice model was established by gavage administration of high dose of Glycyrrhiza uralensis decoction(7.5 g/kg). K~+ concentration in urine and feces of mice was detected. The transcriptome data of intestinal tissues of mice were analyzed to find out the expression changes of genes related to potassium metabolism and altered expression was further confirmed by real-time PCR. The pathological sections of the intestine were observed to reveal the changes of intestinal permeability. The key gene with altered expression was selected as intervention target to confirm its relationship with Glycyrrhiza-induced hypokalemia. Results High dose of Glycyrrhiza uralensis decoction caused hypokalemia in mice. Transcriptome data showed that the expression level of Na~+/K~+ ATPase changed significantly,which was confirmed by real-time PCR. The decreased urinary potassium and the increased K~+ concentration in feces in hypokalemia mice indicated that the hypokalemia induced by Glycyrrhiza uralensis decoction may be mainly caused by intestinal potassium loss. After treatment with Na~+/K~+ ATPase inhibitor,K~+ concentration in urine increased rapidly and K~+ concentration in feces decreased. Continued treatment with high dose of Glycyrrhiza uralensis decoction,K~+ concentration in urine and feces of mice reversed when elimination of Na~+/K~+ ATPase inhibitor. Conclusion Hypokalemia induced by high dose of Glycyrrhiza aqueous extract is mainly caused by gastrointestinal potassium loss,which can be specifically abrogated by Na~+/K~+ ATPase inhibitors.
Objective To explore the molecular mechanism of hypokalemia induced by high dose of Glycyrrhiza uralensis decoction in mice.Methods Hypokalemia mice model was established by gavage administration of high dose of Glycyrrhiza uralensis decoction(7.5 g/kg). K~+ concentration in urine and feces of mice was detected. The transcriptome data of intestinal tissues of mice were analyzed to find out the expression changes of genes related to potassium metabolism and altered expression was further confirmed by real-time PCR. The pathological sections of the intestine were observed to reveal the changes of intestinal permeability. The key gene with altered expression was selected as intervention target to confirm its relationship with Glycyrrhiza-induced hypokalemia. Results High dose of Glycyrrhiza uralensis decoction caused hypokalemia in mice. Transcriptome data showed that the expression level of Na~+/K~+ ATPase changed significantly,which was confirmed by real-time PCR. The decreased urinary potassium and the increased K~+ concentration in feces in hypokalemia mice indicated that the hypokalemia induced by Glycyrrhiza uralensis decoction may be mainly caused by intestinal potassium loss. After treatment with Na~+/K~+ ATPase inhibitor,K~+ concentration in urine increased rapidly and K~+ concentration in feces decreased. Continued treatment with high dose of Glycyrrhiza uralensis decoction,K~+ concentration in urine and feces of mice reversed when elimination of Na~+/K~+ ATPase inhibitor. Conclusion Hypokalemia induced by high dose of Glycyrrhiza aqueous extract is mainly caused by gastrointestinal potassium loss,which can be specifically abrogated by Na~+/K~+ ATPase inhibitors.
引文
[1] 叶菊,蔺海明,邱黛玉,等.几种甘草(Glycyrrhiza)光合特性、形态特征及生物量比较[J].中国沙漠,2014,34(2):456-463.
[2] 孙娟娟.甘草甜素片与低钾血症[J].交通医学,1994(2):139-140
[3] 李翠兰,白春峰,王者令.甘草引起的低钾血症[J].中医杂志,1996(9):570..
[4] 罗晓媛,褚燕琦.复方甘草酸苷注射液致低钾血症1例[J].医药导报,2011,30(01):125-126.
[5] 王敬衔,郑寿焕.甘草片引起的严重低钾血症2例[J].延边大学医学学报,2013,36(4):316-317.
[6] 苏文凌,杨洋,苗芳,等.复方甘草片致低钾血症2例[J].中国医院药学杂志,2015,35(15):1439-1440.
[7] 白华,王璞,张学红,等.复方甘草酸苷片致低钾血症1例[J].传染病信息,2018,31(04):380-382.
[8] WALKER B R,EDWARDS C R.Licorice-induced hypertension and syndromes of apparent mineralocorticoid excess[J].Endocrinol Metab Clin North Am,1994,23(2):359-377.
[9] 李丹丹,刘扬,庞妩燕.复方甘草酸二铵胶囊致重症低钾血症及横纹肌溶解1例及文献复习[J].重庆医学,2015,44(13):1870-1871.
[10] 赖斌,王意,赵亮,等.甘草酸二铵胶囊致长期低钾血症1例[J].医药导报,2018,37(3):393-394.
[11] 李鹏飞,谭九根.严重低钾血症致横纹肌溶解3例临床分析[J].现代实用医学,2010,22(7):784-785.
[12] 雷龙,吴华,刘昕.严重低钾血症致横纹肌溶解综合征1例报告[J].北京医学,2010,32(4):295-296.
[13] 郑杨杨.复方甘草酸苷片致严重低钾血症及横纹肌溶解1例[J].广西医学,2018,40(10):1246,1257..
[14] SHAH M,WILLIAMS C,AGGARWAL A,et al.Licorice-related rhabdomyolysis:a big price for a sweet tooth[J].Clin Nephrol,2012,77(6):491-495.
[15] 何免.甘草水煎剂中miRNA的提取、高通量测序及对免疫细胞的影响研究[D].广州:广东药学院,2015.
[16] 孙敬方.实验动物方法学[M].北京:人民卫生出版社,2001:116
[17] 杜学勤,刘焕蓉,郭志强,等.高效液相色谱法测定水煎法提取中成药中的甘草酸含量[J].时珍国医国药,2007,18(4):812-814.
[18] LIMA D,VALENTE R,CAPELLA M.Ouabain-induced alterations in ABCB1 of mesenteric lymph nodes and thymocytes of rats and mice[J].Oncol Lett,2016,12(6):5275-5280.
[19] 张寒钰.复方甘草酸苷致严重低血钾与横纹肌溶解[J].药物不良反应杂志,2014,16(2):123-124.
[20] CHEN C C,KOLOSOV D,KELLY S P.Effect of the liquorice root derivatives on salt and water balance in a teleost fish,rainbow trout (Oncorhynchus mykiss)[J].Comp Biochem Physiol A Mol Integr Physiol,2015,180:86-97.
[21] CHANG J,LEONG R W,WASINGER V,et al.Impaired intestinal permeability contributes to ongoing bowel symptoms in patients with inflammatory bowel disease and mucosal healing[J].Gastroenterology,2017,153(3):723-731.
[2] 孙娟娟.甘草甜素片与低钾血症[J].交通医学,1994(2):139-140
[3] 李翠兰,白春峰,王者令.甘草引起的低钾血症[J].中医杂志,1996(9):570..
[4] 罗晓媛,褚燕琦.复方甘草酸苷注射液致低钾血症1例[J].医药导报,2011,30(01):125-126.
[5] 王敬衔,郑寿焕.甘草片引起的严重低钾血症2例[J].延边大学医学学报,2013,36(4):316-317.
[6] 苏文凌,杨洋,苗芳,等.复方甘草片致低钾血症2例[J].中国医院药学杂志,2015,35(15):1439-1440.
[7] 白华,王璞,张学红,等.复方甘草酸苷片致低钾血症1例[J].传染病信息,2018,31(04):380-382.
[8] WALKER B R,EDWARDS C R.Licorice-induced hypertension and syndromes of apparent mineralocorticoid excess[J].Endocrinol Metab Clin North Am,1994,23(2):359-377.
[9] 李丹丹,刘扬,庞妩燕.复方甘草酸二铵胶囊致重症低钾血症及横纹肌溶解1例及文献复习[J].重庆医学,2015,44(13):1870-1871.
[10] 赖斌,王意,赵亮,等.甘草酸二铵胶囊致长期低钾血症1例[J].医药导报,2018,37(3):393-394.
[11] 李鹏飞,谭九根.严重低钾血症致横纹肌溶解3例临床分析[J].现代实用医学,2010,22(7):784-785.
[12] 雷龙,吴华,刘昕.严重低钾血症致横纹肌溶解综合征1例报告[J].北京医学,2010,32(4):295-296.
[13] 郑杨杨.复方甘草酸苷片致严重低钾血症及横纹肌溶解1例[J].广西医学,2018,40(10):1246,1257..
[14] SHAH M,WILLIAMS C,AGGARWAL A,et al.Licorice-related rhabdomyolysis:a big price for a sweet tooth[J].Clin Nephrol,2012,77(6):491-495.
[15] 何免.甘草水煎剂中miRNA的提取、高通量测序及对免疫细胞的影响研究[D].广州:广东药学院,2015.
[16] 孙敬方.实验动物方法学[M].北京:人民卫生出版社,2001:116
[17] 杜学勤,刘焕蓉,郭志强,等.高效液相色谱法测定水煎法提取中成药中的甘草酸含量[J].时珍国医国药,2007,18(4):812-814.
[18] LIMA D,VALENTE R,CAPELLA M.Ouabain-induced alterations in ABCB1 of mesenteric lymph nodes and thymocytes of rats and mice[J].Oncol Lett,2016,12(6):5275-5280.
[19] 张寒钰.复方甘草酸苷致严重低血钾与横纹肌溶解[J].药物不良反应杂志,2014,16(2):123-124.
[20] CHEN C C,KOLOSOV D,KELLY S P.Effect of the liquorice root derivatives on salt and water balance in a teleost fish,rainbow trout (Oncorhynchus mykiss)[J].Comp Biochem Physiol A Mol Integr Physiol,2015,180:86-97.
[21] CHANG J,LEONG R W,WASINGER V,et al.Impaired intestinal permeability contributes to ongoing bowel symptoms in patients with inflammatory bowel disease and mucosal healing[J].Gastroenterology,2017,153(3):723-731.