摘要
目的研究制首乌诱导肝细胞凋亡的"毒-证"机制。方法皮下注射氢化可的松建立大鼠肾阳虚模型,分为空白对照组、肾阳虚低、中、高剂量组(分别予制首乌10.8,21.6,43.2 g/kg)。连续给药14 d后处死大鼠,制备含药血清。分别采用5,5'-二硫代双(2-硝基苯甲酸)(DTNB)法和硫代巴比妥酸(TBA)法测定L02肝细胞内谷胱甘肽(GSH)和丙二醛(MDA)的含量,并通过显微镜和电镜观察肝细胞形态结构及超微结构的改变。结果制首乌肾阳虚各剂量组含药血清(终浓度15%)作用24 h及48 h后,细胞内T-GSH及GSH含量均有不同程度降低,其中肾阳虚中、高剂量组含药血清作用48 h后,细胞内GSH含量与空白对照组比较有统计学差异(P<0.05);同时,GSH/GSSG比值与空白组比较,制首乌肾阳虚高剂量组含药血清作用24 h及48 h后均出现显著降低(P<0.01)。给药后,L02肝细胞密度变小,脱壁细胞增多,部分细胞出现坏死、裂解和形态改变;肝细胞超微结构也发生变化,胞质出现空泡,细胞器肿胀、破坏。结论制首乌非对证治疗肾阳虚证候时,对肝脏具有一定毒性作用,肾阳虚证可能是制首乌肝毒性的靶向证候,故临床使用时应注意对证用药,并严格控制制首乌的使用剂量和疗程。
Objective To study the mechanisms related ＂ toxic-targeted syndrome＂ of medicated serum of Polygonum Multiflorum Thunb（ PMT） on apoptosis of L02 cells. Methods Rat models with deficiency of kidney-YANG were induced by hydrocortisone. The rats were divided into blank control,low dose（ 10. 8 g / kg）,middle dose（ 21. 6 g / kg）,high dose（ 43. 2 g / kg） groups of PMT,the rats were killed after administrated 14 d to prepare medicated serum of PMT. The contents of glutathione（ GSH） and malondialdehyde（ MDA） in L02 cells were tested by dithio-bis-nitrobenzoic acid（ DTNB） and 2-Thiobarbituric acid（ TBA）. The changes of morphology and ultrastructure of L02 cells were observed by microscope and electron microscope. Results Medicated serum of deficiency of kidney-Yang groups（ 15%） obviously reduced the contents of T-GSH and GSH in L02 hepatocytes after treated 24 h and 48 h. The contents of T-GSH and GSH were differences between control group and high and middle dose groups（ P 0. 05）. Meanwhile,compared with that of control group,the GSH / GSSG ratio of medicated serum of deficiency of kidney-Yang groups were both significantly reduced after treated 24 h and 48 h（ P 0. 01）. L02 liver cell density was decreased,and more cells falling down from the inner wall of the cell culture flasks,and part of the cell became necrotic,lysis and changed morphology. The ultrastructures of L02 liver cell were also changed,such as empty cytoplasm and swelling organelle appeared. Conclusions The syndrome of deficiency of kidney-YANG might be targeting the liver toxicity syndromes of PMT. PMT should be used in the correct syndrome in clinical and strictly control clinical dose and duration of treatment.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2016年第2期262-265,共4页
Chinese Journal of Gerontology
基金
国家自然科学基金面上项目(81373988)
关键词
制首乌
肝细胞
凋亡
Polygonum multiflorum thunb
Hepatocyte
Apoptosis
