基于大黄蒽醌、鞣质类物质作用于HK-2细胞的毒效相关性研究
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摘要
目的：本课题拟以临床广泛应用,同时毒效双重作用客观存在的大黄为示范药物,主要以中药毒效相关性影响因素中量的因素及不同状态因素为研究内容,考察大黄蒽醌和鞣质类物质对HK-2细胞的毒效作用,毒效物质基础,影响因素及作用机制,同时为其它有毒中药的毒效相关性研究提供借鉴。
方法：首先采用正常HK-2细胞,对大黄主要有效部位蒽醌及鞣质,蒽醌部位代表性单体大黄素,鞣质部位代表性类单体的聚合物EGCG的毒性进行研究,通过细胞形态学观察,细胞增殖活性测定,细胞LDH漏出量测定,细胞周期分析及凋亡率测定考察其毒性及量-毒关系。其次,建立细胞损伤模型即以过氧化氢损伤HK-2细胞,在此基础上探讨大黄蒽醌及鞣质、大黄素、EGCG对于细胞氧化应激损伤的保护作用,通过细胞形态学观察,细胞增殖活性测定,细胞LDH测定,细胞周期分析及凋亡率测定,考察其量-(毒/效)关系。最后综合分析大黄蒽醌、鞣质类物质对HK-2细胞的毒效物质基础,影响因素及作用机制。
结果：
(1)大黄蒽醌及大黄素总体上可使细胞形态发生皱缩,融合等形态学的变化；大黄蒽醌和大黄素使细胞生长受到抑制,阻滞了细胞周期的正常转化,并不同程度的引起了细胞的凋亡和坏死。表明大黄蒽醌和大黄素对HK-2细胞有潜在的肾毒性,能够导致细胞的损伤。大黄蒽醌对HK-2细胞增殖有直接的抑制作用,呈一定的量效关系,半抑制浓度约为20.62mg·L-1；大黄素显现的毒性作用较轻,半抑制浓度约为120μmol·L-1。大黄蒽醌和大黄素的毒性作用机制主要是阻滞了细胞周期从S期向G2/M期的转化,并引起细胞的凋亡和坏死。
(2)大黄鞣质部位在考察剂量中对HK-2细胞的毒性作用轻微,对于细胞形态的影响只是轻微的使细胞形状变得狭长,抑制率在较高浓度200mg·L-1时仅达到20%,而大黄蒽醌的半抑制浓度20.62 mg·L-1,蒽醌类物质和鞣质类物质在大黄生药中所占的质量百分比约为1：2左右,因此,在应用大黄时鞣质的肾细胞毒性相对于蒽醌类物质有限。鞣质的单体的聚合物EGCG对HK-2细胞的毒性作用较轻,在80μmol·L-1以上的浓度可引起HK-2细胞早期凋亡。
(3)HK-2细胞在过氧化氢损伤的状态下,一方面,大黄蒽醌主要表现出了对受损细胞的保护作用,如细胞形态上相对于损伤组细胞的形态有所恢复,细胞活性有所增强。尤其是在LDH的检测结果中,蒽醌组的光密度值明显减少。比较有意义的是,在凋亡率的检测中,蒽醌组的细胞凋亡率相对于损伤组有明显改善,近一步证实蒽醌类物质对受损细胞有保护作用。而另一方面,从剂量关系上看,15mg·L-1以下剂量范围内的大黄蒽醌对于受损细胞有一定的保护作用；当剂量达到30mg-L-1时,大黄蒽醌一方面发挥了细胞保护作用,同时也显示了一定的潜在毒性作用,但损伤细胞对大黄蒽醌的毒性耐受性比正常状态下增强。大黄素对过氧化氢损伤HK-2细胞的作用中,并未发现有明显的保护作用,尤其在对凋亡率的检测中,发现大黄素引起细胞晚期凋亡/坏死的作用显著,比大黄素对正常细胞的晚期凋亡/坏死的影响更为明显,提示在HK-2细胞损伤状态下,大黄素更多的体现了其毒性作用。
(4)HK-2细胞在过氧化氢损伤的状态下,相同剂量大黄鞣质相对于对正常状态下细胞的毒性,其毒性作用变化不大,细胞形态改变不明显,细胞增殖活性没有显著下降；在细胞损伤状态下,大黄鞣质在100mg.L-1时对细胞的抑制率为27%,与该剂量对正常状态细胞的抑制率22%相差不大。尤其是LDH的检测中,相对于损伤组,LDH值下降明显,表明大黄鞣质对受损细胞具有保护作用,受损细胞对鞣质毒性的耐受性增加。在本实验中,EGCG对损伤的HK-2细胞表现了一定的保护作用,尤其对于凋亡率的影响上,早期凋亡和晚期凋亡/死亡的改善作用明显。
结论：
基于上述对大黄毒效辩证关系的认识,结合中药毒效相关性观点的科学内涵,提出对于有毒中药的毒效相关性研究应考察以下两个方面：剂量是有毒中药毒效相关性的影响因素；应在机体生理、病理不同状态下进行有毒中药毒效相关性研究。以大黄为例,蒽醌和鞣质类物质对HK-2细胞的毒效相关性主要体现在以下几个方面：
(1)成分不同,毒性相异：蒽醌类物质(蒽醌有效部位和大黄素)的毒性作用强于鞣质类物质(鞣质有效部位和EGCG),主要影响了HK-2细胞形态、降低细胞增殖活性、增加细胞LDH的漏出、阻滞细胞周期、引起细胞的凋亡/坏死等。
(2)成分不同,药效相似：HK-2细胞在过氧化氢损伤状态下,蒽醌类物质(主要是蒽醌有效部位)和鞣质类物质(主要是鞣质有效部位和EGCG)在一定剂量范围内都发挥了药效作用,如保护细胞形态,提高细胞增殖活性,降低细胞凋亡率等。
(3)成分相同,机体状态有别,量-毒/效关系不同：相对于正常状态下,大黄蒽醌和鞣质对过氧化氢损伤的HK-2细胞的毒性作用降低,损伤细胞对药物毒性的耐受性增强,在相同剂量下主要体现了药效作用；相对于正常状态,大黄素对损伤的HK-2细胞毒性作用明显,EGCG则主要体现对损伤HK-2细胞的药效作用。
(4)成分相同,用量有别,量-毒关系不同：如：对于正常细胞大黄蒽醌在30mg·L-1以下剂量时,其抑制细胞增殖作用和对细胞周期的阻滞作用相对较弱,在30mg·L-1增加到60mg·L-1时的过程中,抑制细胞增殖和细胞周期的阻滞作用增加明显,而且出现了明显的晚期凋亡/坏死,晚期凋亡/坏死程度在60mg·L-1时甚至大于了50%,毒性作用明显。
(5)通过考察大黄蒽醌、鞣质类物质对HK-2细胞的毒效关系,为其它中药的毒效相关性研究可提供思路和方法借鉴。
Objective The goal of this thesis is to explore correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb, and the influencing factors of dose and different conditions of HK-2 cells are to be concerned about, what is more significant, this study may provide demonstration for researches on correlation between toxicity and effect of other toxic TCM.
Methods The study on toxicity of anthraquinone and tannin extracted from Rhubarb, emodin and EGCG on HK-2 cells was carried out in different dose, and the change of cell morphology, OD of LDH, cell cycle and apoptosis were detected. Furthermore, HK-2 cells that were damaged at some level by exposing to H2O2 were pretreated with anthraquinone and tannin extracted from Rhubarb, emodin and EGCG,the effect of those drugs on impaired HK-2 cells were detected. By combinding with experimental data and traditional theory of TCM, the material basic, influencing factors and mechanism of correlation between toxicity and effct of Rhubarb were analyzed.
Results (1) Anthraquinone extracted from Rhubarb and emodin possed some toxic on HK-2 cells, the toxic manifestation included:inhibition of cells proliferation, cell shrinkage and vacuolation, LDH leakage were increased, the cell cycle was blocked and induced apoptosis at different level. This result illustrated anthraquinone and emodin may be the materials that damage the kidney. The IC50 of anthraquinone extracted from Rhubarb is about 20.62mg·L-1 and the IC50 of emodin is about 120μmol·L-1. The main toxic mechanism of anthraquinone extracted from Rhubarb and emodin is to induce apoptosis and block the transformation of cell cycle from S period to G2/M period.
(2) The toxicity of tannin extracted from Rhubarb on HK-2 cell was little, only leaded to lower grade change of cell shape, the inhibition ratio was 20% when the dose was 200mg·L-1.The percentage of anthraquinone and tannin extracted from Rhubarb is 1:2 approximately, contrast to the toxicity of anthraquinone, tannin is safe when the Rhubarb is administrated. The early apoptosis was induced by using EGCG when the dose exceed 80μmol·L-1.
(3) In the condition of HK-2 cells were damaged by H2O2, anthraquinone extracted from Rhubarb protected the cells by improving the cell shape and activity, the OD was descended, especially, the apoptosis of cell was improved significantly. The effect of anthraquinone was concerned with the dose,15mg·L-1 anthraquinone mainly protected the cell, toxicity was detected when the dose was 30 mg·L-1, but the tolerance to drug toxicity of damaged cells was enhanced. Emodin represented the toxicity mainly by inducing advanced stage apoptosis and necrosis.
(4) In the condition of HK-2 cell were damaged by H2O2, tannin extracted from Rhubarb protected the cells and the tolerance of damaged cells to drug toxicity was enhanced. The inhibitor ratio changed lighty and the OD of LDH was descended. EGCG protected the damaged cell by improving the early stage apoptosis and necrosis.
Conclusions Base on the experiment on the toxicity and effct of Rhubarb, combind with the idea of correlation between toxicity and effect of toxic TCM, the thesis provide the strategy to the research on correlation between toxicity and effect of toxic TCM:The influencing factors of correlation between toxicity and effect are varied, but dose and different conditions play important roles. The correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb include some aspects as follows:
(1) Different materials may have different toxicity. Anthraquinone extracted from Rhubarb and emodin are more toxical than tannin extracted from Rhubarb and EGCG.. Anthraquinone extracted from Rhubarb and emodin change the cell shape, enhance the inhibition ratio and OD of LDH, block the cell cycle and induce the apoptosis and necrosis.
(2) Different materials may have similar effect. Anthraquinone extracted from Rhubarb and emodin all protect the HK-2 cells damaged by H2O2. They all can maintain the normal cell shape, descend the inhibition ratio and cell apoptosis et al.
(3) The same materials may have different toxicity and effect in different conditions. The tolerance of HK-2 cells to toxicity of anthraquinone and tannin extracted from Rhubarb are enhanced, anthraquinone and tannin extracted from Rhubarb mainly protect the damaged cells contrast to the normal condition at the same dose. The toxicity of emodin on damaged cells is more significant than the normal condition but EGCG protect the damaged cells.
(4) The same materials may have different toxicity and effect in different dose. The toxicity of anthraquinone extracted from Rhubarb is little when the dose is lower than 30mg·L-1 and enhanced significantly when the dose is 60 mg·L-1, the inhibition ratio, the apoptosis and necrosis of HK-2 cells alter notablely.
(5) The study may provide reference to explore correlation between toxicity and effect of other toxic TCM.
方法：首先采用正常HK-2细胞,对大黄主要有效部位蒽醌及鞣质,蒽醌部位代表性单体大黄素,鞣质部位代表性类单体的聚合物EGCG的毒性进行研究,通过细胞形态学观察,细胞增殖活性测定,细胞LDH漏出量测定,细胞周期分析及凋亡率测定考察其毒性及量-毒关系。其次,建立细胞损伤模型即以过氧化氢损伤HK-2细胞,在此基础上探讨大黄蒽醌及鞣质、大黄素、EGCG对于细胞氧化应激损伤的保护作用,通过细胞形态学观察,细胞增殖活性测定,细胞LDH测定,细胞周期分析及凋亡率测定,考察其量-(毒/效)关系。最后综合分析大黄蒽醌、鞣质类物质对HK-2细胞的毒效物质基础,影响因素及作用机制。
结果：
(1)大黄蒽醌及大黄素总体上可使细胞形态发生皱缩,融合等形态学的变化；大黄蒽醌和大黄素使细胞生长受到抑制,阻滞了细胞周期的正常转化,并不同程度的引起了细胞的凋亡和坏死。表明大黄蒽醌和大黄素对HK-2细胞有潜在的肾毒性,能够导致细胞的损伤。大黄蒽醌对HK-2细胞增殖有直接的抑制作用,呈一定的量效关系,半抑制浓度约为20.62mg·L-1；大黄素显现的毒性作用较轻,半抑制浓度约为120μmol·L-1。大黄蒽醌和大黄素的毒性作用机制主要是阻滞了细胞周期从S期向G2/M期的转化,并引起细胞的凋亡和坏死。
(2)大黄鞣质部位在考察剂量中对HK-2细胞的毒性作用轻微,对于细胞形态的影响只是轻微的使细胞形状变得狭长,抑制率在较高浓度200mg·L-1时仅达到20%,而大黄蒽醌的半抑制浓度20.62 mg·L-1,蒽醌类物质和鞣质类物质在大黄生药中所占的质量百分比约为1：2左右,因此,在应用大黄时鞣质的肾细胞毒性相对于蒽醌类物质有限。鞣质的单体的聚合物EGCG对HK-2细胞的毒性作用较轻,在80μmol·L-1以上的浓度可引起HK-2细胞早期凋亡。
(3)HK-2细胞在过氧化氢损伤的状态下,一方面,大黄蒽醌主要表现出了对受损细胞的保护作用,如细胞形态上相对于损伤组细胞的形态有所恢复,细胞活性有所增强。尤其是在LDH的检测结果中,蒽醌组的光密度值明显减少。比较有意义的是,在凋亡率的检测中,蒽醌组的细胞凋亡率相对于损伤组有明显改善,近一步证实蒽醌类物质对受损细胞有保护作用。而另一方面,从剂量关系上看,15mg·L-1以下剂量范围内的大黄蒽醌对于受损细胞有一定的保护作用；当剂量达到30mg-L-1时,大黄蒽醌一方面发挥了细胞保护作用,同时也显示了一定的潜在毒性作用,但损伤细胞对大黄蒽醌的毒性耐受性比正常状态下增强。大黄素对过氧化氢损伤HK-2细胞的作用中,并未发现有明显的保护作用,尤其在对凋亡率的检测中,发现大黄素引起细胞晚期凋亡/坏死的作用显著,比大黄素对正常细胞的晚期凋亡/坏死的影响更为明显,提示在HK-2细胞损伤状态下,大黄素更多的体现了其毒性作用。
(4)HK-2细胞在过氧化氢损伤的状态下,相同剂量大黄鞣质相对于对正常状态下细胞的毒性,其毒性作用变化不大,细胞形态改变不明显,细胞增殖活性没有显著下降；在细胞损伤状态下,大黄鞣质在100mg.L-1时对细胞的抑制率为27%,与该剂量对正常状态细胞的抑制率22%相差不大。尤其是LDH的检测中,相对于损伤组,LDH值下降明显,表明大黄鞣质对受损细胞具有保护作用,受损细胞对鞣质毒性的耐受性增加。在本实验中,EGCG对损伤的HK-2细胞表现了一定的保护作用,尤其对于凋亡率的影响上,早期凋亡和晚期凋亡/死亡的改善作用明显。
结论：
基于上述对大黄毒效辩证关系的认识,结合中药毒效相关性观点的科学内涵,提出对于有毒中药的毒效相关性研究应考察以下两个方面：剂量是有毒中药毒效相关性的影响因素；应在机体生理、病理不同状态下进行有毒中药毒效相关性研究。以大黄为例,蒽醌和鞣质类物质对HK-2细胞的毒效相关性主要体现在以下几个方面：
(1)成分不同,毒性相异：蒽醌类物质(蒽醌有效部位和大黄素)的毒性作用强于鞣质类物质(鞣质有效部位和EGCG),主要影响了HK-2细胞形态、降低细胞增殖活性、增加细胞LDH的漏出、阻滞细胞周期、引起细胞的凋亡/坏死等。
(2)成分不同,药效相似：HK-2细胞在过氧化氢损伤状态下,蒽醌类物质(主要是蒽醌有效部位)和鞣质类物质(主要是鞣质有效部位和EGCG)在一定剂量范围内都发挥了药效作用,如保护细胞形态,提高细胞增殖活性,降低细胞凋亡率等。
(3)成分相同,机体状态有别,量-毒/效关系不同：相对于正常状态下,大黄蒽醌和鞣质对过氧化氢损伤的HK-2细胞的毒性作用降低,损伤细胞对药物毒性的耐受性增强,在相同剂量下主要体现了药效作用；相对于正常状态,大黄素对损伤的HK-2细胞毒性作用明显,EGCG则主要体现对损伤HK-2细胞的药效作用。
(4)成分相同,用量有别,量-毒关系不同：如：对于正常细胞大黄蒽醌在30mg·L-1以下剂量时,其抑制细胞增殖作用和对细胞周期的阻滞作用相对较弱,在30mg·L-1增加到60mg·L-1时的过程中,抑制细胞增殖和细胞周期的阻滞作用增加明显,而且出现了明显的晚期凋亡/坏死,晚期凋亡/坏死程度在60mg·L-1时甚至大于了50%,毒性作用明显。
(5)通过考察大黄蒽醌、鞣质类物质对HK-2细胞的毒效关系,为其它中药的毒效相关性研究可提供思路和方法借鉴。
Objective The goal of this thesis is to explore correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb, and the influencing factors of dose and different conditions of HK-2 cells are to be concerned about, what is more significant, this study may provide demonstration for researches on correlation between toxicity and effect of other toxic TCM.
Methods The study on toxicity of anthraquinone and tannin extracted from Rhubarb, emodin and EGCG on HK-2 cells was carried out in different dose, and the change of cell morphology, OD of LDH, cell cycle and apoptosis were detected. Furthermore, HK-2 cells that were damaged at some level by exposing to H2O2 were pretreated with anthraquinone and tannin extracted from Rhubarb, emodin and EGCG,the effect of those drugs on impaired HK-2 cells were detected. By combinding with experimental data and traditional theory of TCM, the material basic, influencing factors and mechanism of correlation between toxicity and effct of Rhubarb were analyzed.
Results (1) Anthraquinone extracted from Rhubarb and emodin possed some toxic on HK-2 cells, the toxic manifestation included:inhibition of cells proliferation, cell shrinkage and vacuolation, LDH leakage were increased, the cell cycle was blocked and induced apoptosis at different level. This result illustrated anthraquinone and emodin may be the materials that damage the kidney. The IC50 of anthraquinone extracted from Rhubarb is about 20.62mg·L-1 and the IC50 of emodin is about 120μmol·L-1. The main toxic mechanism of anthraquinone extracted from Rhubarb and emodin is to induce apoptosis and block the transformation of cell cycle from S period to G2/M period.
(2) The toxicity of tannin extracted from Rhubarb on HK-2 cell was little, only leaded to lower grade change of cell shape, the inhibition ratio was 20% when the dose was 200mg·L-1.The percentage of anthraquinone and tannin extracted from Rhubarb is 1:2 approximately, contrast to the toxicity of anthraquinone, tannin is safe when the Rhubarb is administrated. The early apoptosis was induced by using EGCG when the dose exceed 80μmol·L-1.
(3) In the condition of HK-2 cells were damaged by H2O2, anthraquinone extracted from Rhubarb protected the cells by improving the cell shape and activity, the OD was descended, especially, the apoptosis of cell was improved significantly. The effect of anthraquinone was concerned with the dose,15mg·L-1 anthraquinone mainly protected the cell, toxicity was detected when the dose was 30 mg·L-1, but the tolerance to drug toxicity of damaged cells was enhanced. Emodin represented the toxicity mainly by inducing advanced stage apoptosis and necrosis.
(4) In the condition of HK-2 cell were damaged by H2O2, tannin extracted from Rhubarb protected the cells and the tolerance of damaged cells to drug toxicity was enhanced. The inhibitor ratio changed lighty and the OD of LDH was descended. EGCG protected the damaged cell by improving the early stage apoptosis and necrosis.
Conclusions Base on the experiment on the toxicity and effct of Rhubarb, combind with the idea of correlation between toxicity and effect of toxic TCM, the thesis provide the strategy to the research on correlation between toxicity and effect of toxic TCM:The influencing factors of correlation between toxicity and effect are varied, but dose and different conditions play important roles. The correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb include some aspects as follows:
(1) Different materials may have different toxicity. Anthraquinone extracted from Rhubarb and emodin are more toxical than tannin extracted from Rhubarb and EGCG.. Anthraquinone extracted from Rhubarb and emodin change the cell shape, enhance the inhibition ratio and OD of LDH, block the cell cycle and induce the apoptosis and necrosis.
(2) Different materials may have similar effect. Anthraquinone extracted from Rhubarb and emodin all protect the HK-2 cells damaged by H2O2. They all can maintain the normal cell shape, descend the inhibition ratio and cell apoptosis et al.
(3) The same materials may have different toxicity and effect in different conditions. The tolerance of HK-2 cells to toxicity of anthraquinone and tannin extracted from Rhubarb are enhanced, anthraquinone and tannin extracted from Rhubarb mainly protect the damaged cells contrast to the normal condition at the same dose. The toxicity of emodin on damaged cells is more significant than the normal condition but EGCG protect the damaged cells.
(4) The same materials may have different toxicity and effect in different dose. The toxicity of anthraquinone extracted from Rhubarb is little when the dose is lower than 30mg·L-1 and enhanced significantly when the dose is 60 mg·L-1, the inhibition ratio, the apoptosis and necrosis of HK-2 cells alter notablely.
(5) The study may provide reference to explore correlation between toxicity and effect of other toxic TCM.
引文
[1]于智敏,吕爱平,吴萍等.对中药安全性评价研究的思考[J].中国中医基础医学杂志,2003,9(5)：66-68.
[2]刘树民,罗明媚,李玉洁.中药毒性理论及研究进展[J].世界科学技术-中药现代化,2003,5(3)：45-47.
[3]王峥涛.中药毒性本质的科学研究[J].中药药性理论继承与创新研究简报,2007,(1)：16-20.
[4]肖小河,金城,王伽伯,等.中医药学科发展报告[M].北京：中国科学技术文献出版社,2006.
[5]彭成,王昌恩,林娜.有毒中药毒效相关性基础研究的意义与实践[J].中药药理与临床,2008,24(1)：71-73.
[6]雷载权,张廷模.中华临床中药学[M].北京：人民卫生出版社,1998：143.
[7]常惟智.影响中药功效及作用方向选择性之相关因素探析[J].中医药学报,2005,33(2)：12-13.
[8]余葱葱,郭力,彭成.毒效双组分控制有毒中药附子临床单煎工艺研究[J].时珍国医国药,2009,20(12)：3087-3089.
[9]王嫣虹,彭成,朱力阳.大黄素对结肠Cajal间质细胞毒效作用的研究[J].中国中西医合消化杂志,2010,18(3)：141-144.
[10]阮雯聪.影响中药功效与毒副作用的因素[J].浙江中西医结合杂志,2008,18(3)：197-198.
[11]由凤鸣,贾波,邓中甲.从剂量对中药功效发挥方向的影响论中药的矢量性[J].辽宁中医药大学学报.2010,12(5)：120-121.
[12]National Toxicology Program,2001. Technical report on the toxicology and carcinogenesis studies of emodin in F344/N rats and B6C3FI mice. Series No.493. NIH Publication No. 01-3952. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC.
[13]王伽伯,马永刚,张萍.炮制对大黄化学成分和肝肾毒性的影响及其典型相关分析[J].药学学报,2009,44(8)：885-890.
[14]谭余庆.对大黄素及大黄蒽醌化合物肾毒性和致癌性的再认识.国家药品审评中心电子刊物.
[15]Wang CF, Wu XD, Chen M,. Emodin induces apoptosis through caspase 3-dependent pathway in HK-2 cells[J]. Toxicology,2007,231(2-3):120-128.
[16]Yan M, Zhang LY, Sun LX, Jiang ZZ, Xiao XH. Nephrotoxicity study of total rhubarb anthraquinones on Sprague Dawley rats using DNA microarrays[J]. Journal of Ethnopharmacology,2006,107(2):308-311.
[17]张陆勇,江振洲,蒲存海.大黄总蒽醌对SD大鼠灌胃给药的长期毒性研究[J].中国生化药物杂志,2004,25(4)：206-209.
[18]Mueller SO, Stopper H., Dekant W. Biotransformation of the anthraquinones emodin and chrysophanol by cytochrome P450 enzymes:Bioactivation to genotoxic metabolites [J]. Drug Metabolism and Disposition,1998,26:540-546.
[19]Xiaomei Li, Haiyan Wang. Chinese herbal medicine in the treatment of chronic kidney disease [J]. Advances in Chronic Kidney Disease,2005,12(3):276-281.
[20]肖炜,邓虹珠,马云.大黄治疗慢性肾功能衰竭的临床与实验研究概述[J].中国中药杂志,2002,27(4)：241-262.
[21]Mitsuma, T., Terasawa, K., Yokozawa, T., Oura, H. Rhubarb therapy in patients with chronic renal failure (Part 1) [J]. Journal of Medical and Pharmaceutical Society for WAKAN-YAKU,1984,1:266-278.
[22]Mitsuma, T., Yokozawa, T., Oura, H., Terasawa, K. Rhubarb therapy in patients with chronic renal failure (Part 2)[J]. Japanese Journal of Nephrology,1987,29:195-207.
[23]李孜,卿平,冀玲.大黄治疗慢性肾功能衰竭的系统评价[J].中国循证医学杂志,2004,4(7)：468-473.
[24]Leishi Li. End-stage renal disease in China[J]. Kidney International,1996,49:287-301.
[25]肖相如.大黄在慢性肾衰中的正确运用[J].辽宁中医杂志,2002,29(6)：311-312.
[26]Jiabo Wang, Yanling Zhao, Xiaohe Xiao. Assessment of the renal protection and hepatotoxicity of rhubarb extract in rats[J]. Journal of Ethnopharmacology.124 (2009) 18-25.
[27]周本宏,王慧媛,郭志磊.石榴皮鞣质对羟自由基和超氧阴离子自由基的清除作用[J].国医院药学杂志,2008,28(17)：1442-1445.
[28]明玉,倪为为.大黄中鞣质成分的分离与液相色谱/质谱联用分析[J].色谱,2004, 22(6)：605-608.
[29]焦西英,杨浩,钱新宏.EGCG对过氧化氢所致神经元过氧化损伤的保护作用[J].解剖学研究,2008,30(6)：401-404.
[30]Pamela Pedrielli, Laura M. Holkeri, Leif H. Skibsted.Antioxidant activity of (+)-catechin. Rate constant for hydrogen-atom transfer to peroxyl radicals [J]. Eur Food Res Technol (2001)213:405-408.
[31]Yohei Miyamoto, Mariko Sano. Johnl.Haylor. (EGCG)-induced apoptosis in normal rat kidney interstitial fibroblast(NRK-49F)cells [J]. Eur Food Res Technol (2001) 213: 405-408.
[32]邓勇,黄珀,陈同良.肾小管细胞氧化性损伤模型的建立[J].现代泌尿外科杂志,2004,9(3)：144-145
[33]叶俊生.左卡尼汀在肾小管上皮细胞氧化应激损伤中的保护作用研究[D].南方医科大学博士论文,2010.
[34]叶任高,陆再英.内科学第五版[M].北京：人民卫生出版社,2001：569.
[35]万德光,彭成,赵军宁.四川道地中药材志[M].成都：四川科学技术出版社,2005：94.
[36]孙玉琦,肖小河,马永刚.大黄煎煮过程中蒽醌类成分动态变化规律研究[J].解放军药学学报,2006,22：281-283.
[37]杨洋,张蔚,黄丽琼.槲皮素联合顺铂对宫颈癌HeLa细胞增殖及凋亡的影响[J].武汉大学学报(医学版),2009,30(5)：334-336.
[38]邓生琼,周显琎,王瑶.硫化氢对氧化损伤心肌细胞凋亡的影响[J].武汉大学学报(医学版).2009,30(1)：61-65.
[39]李瑞萍,胡雪梅,吕银凤.苦参碱对人卵巢癌细胞增殖的影响及其机制[J].西安交通大学学报(医学版).2010,31(5)：621-624.
[40]孙沂,樊宏伟,王书奎.氯吡格雷对人胃黏膜上皮细胞株GES-1增殖作用的影响[J].世华人消化杂志,2010,18(4)：329-334.
[41]胡良平.现代统计学与SAS应用[M].北京：军事医学科学出版社,2000：367.
[42]刘珺,徐选福,杨文娟.红花注射液对肝星状细胞HSC-T6增殖、凋亡凋亡相关基因表达的影响[M].中草药,2009,40(8)：1270-1274.
[43]Parikh CR, Edelstein CL, Devarajan P. Biomarkers of acute kidney injury:early diagnosis, Pathogenesis, and recovery [J]. J Investig Med.2007,55(7):333-40.
[44]Djamali, A. Oxidative stress as a common Pathway to chronic tubulointerstitial injury in kidney allografts [J]. Am. J. Physiol. Renal Physiol.293:445-455; 2007.
[45]曹纯章,杨绍娟,卜丽莎.过氧化氢诱导诱导心肌细胞凋亡性和非凋亡性死亡的研究[J].中国病理生理杂志,2000,16(5)：457-461.
[46]Thannickal VJ, Fanburg BL. Reactive oxygen species in cell signaling [J]. Am J Physiol Lung Cell Mol Physiol,2000,279(6):L1005-L1028.
[47]McCord JM:Human disease, free radicals, and the oxidant/antioxidant balance [J]. Clin Biochem,1993,26(5):3511-3517.
[48]Vaziri ND, Oveisi F, Ding Y. Role of increased oxygen free radical activity in the Pathogenesis of uremic hypertension [J]. Kidney Int.1998,53:1748-1754.
[49]TePel M, Echelmeyer M, Orie NN,et al. Increased intracellular reactive oxygen Species in Patients with end-stage chronic renal failure:effect of hemodialysis[J].Kidney Int.2000,58:867-872.
[50]Parthasarathy S, Santanam N, Ramachandran S,etal.Oxidants and antioxidant in atherogenesis:an appraisal [J]. Lipid Res.1999,40:2143-2157.
[51]Baliga R, Ueda N, Walker PD, et al. Oxidant mechanisms in toxic renal failure [J]. Drug Metab Rev,1999,31(4):971-997.
[52]Valko,M., Leibfritz,D., Moncol, J.et al. Free radicals and antioxidants in normal physiological functions and human disease [J]. Int.J.Biochem. Cell.Biol.39:44-84;2007.
[53]崔瑛,许伟,海霞.试论中药毒的性能特征[J].河南中医,1997,17(5)：313-315.
[54]张廷模.临床中药学[M].北京：中国中医药出版社,2004,63.
[55]杨藻宸.医学药理学[M].北京：人民卫生出版社,1994：48.
[56]万德光.中药品种品质与药效[M].上海：上海科学技术出版社,2006：94.
[57]王筠默.中药药理学[M].上海：上海科学技术出版社,1985：12.
[58]李萌,王岩.乐陵大枣与静海金丝小枣药理作用的比较[J].中药材,1993,16(6)：35.
[59]聂淑琴,李兰芳,杨庆.5种产地苍术提取物主要药理作用比较研究[J].中国实验方剂学杂志,2002(81)：358-363.
[60]阴健,郭力弓.中药现代研究与临床应用[M].北京：学苑出版社,1993：4,6,24,596.
[61]徐楚江.中药炮制学[M].上海：上海科学技术出版社,1985：84.
[62]李仪奎,姜名瑛.中药药理学[M].北京：中国中医药山版社,1992：17.
[63]Yi Qing, Jia-bo Wang, Wei-jun Kong, et al. The diarrhoeogenic and antidiarrhoeal bidirectional effects of rhubarb and its potential mechanism [J]. Journal of Ethnopharmacology 133 (2011) 1096-1102.
[64]赵红玉,周春祥.基于药、毒效成分辩证关系探讨有毒中药研究思路[J].中草药,2008,39(11)：1601-1603.
[65]彭成.中医药动物实验方法学[M].北京：人民卫生出版社,2008：1223.
[66]王志鹏,梅其炳,刘琳娜.庆大霉素对人肾小管上皮细胞系(HK-2)热休克蛋白(HSP)70表达的影响[J].中国药理学通报,2006,22(4)：497-500.
[67]周桂凤,刘栋,乔玉峰.铅对体外培养HK-2细胞毒性及碘化钾的拮抗作用[J].毒理学杂志,2008,22(4)：283-286.
[68]唐功耀,田雪飞.马兜铃酸对人肾细胞作用的实验研究[J].中华肾脏病杂志,2002,18(4)：266-269.
[69]李彪,李晓玫,张翠英.马兜铃内酞胺对肾小管上皮细胞的损伤作用[J],中国中药杂志,2004：28(1)：79-83.
[70]王清秀.大黄及其主要成份的毒性毒理研究.军事医学科学院博士学位论文[D].2007.
[71]王清秀,吴纯启,杨红莲.大黄素对HK-2细胞周期增殖的影响毒理学杂志[J].2007,21(6)：440-444.
[72]李瑞萍,胡雪梅,吕银凤.苦参碱对人卵巢癌细胞增殖的影响及其机制[J].西安交通大学学报,2010,31(5)：621-623.
[73]卢薇娜.辛伐他汀和血脂康对糖尿病大鼠肾脏细胞凋亡的影响及其可能机制[D].2010.
[74]彭黎明,王曾礼.细胞凋亡的基础与临床[M].北京：人民卫生出版社,2000：2.
[75]王彩芳.鞣质生理活性研究进展[J].国外医学中医中药分册,2001,23(5)：278-282.
[76]Yoshioka H, Akai G, Yoshinage K. Protecting effect of a green tea percolate and its main constituents against gamma ray-induced scission of DNA[J]. Biosci Biotech Biochem, 1996,60:117-119.
[77]Yang GY, Liao J, Kim K, etal. Inhibition of growth and induction of apoptosis in human caneer cell lines by tea polyphenols[J]. Carcinogenesis,1998:19:611-616.
[78]Liang YC, Shoei L S. Lin JK, et al. Inhibition of cyclin-dependent kinases 2 and 4 activities as well as induction of cdk inhibitors P21 and P27 during growth arrest of human breast carcinoma cells by(-)-epigallocatechin-3-gallate[J]. J Cellular biochem.1999,75:1-8.
[79]Yohei MAYAMOTO, John.L HAYLOR, A.Meguid EL NAHAS. Cellular toxicity of catechin analogues containing gallte in opossum kidney Proximal tubular(OK) cells[J]. The Journal of toxicological sciences,29(1):47-52,2004.
[1]万德光.中药品种品质与药效[M].上海：上海科学技术出版社,2007：164-165.
[2]李文渊,童丽,等.唐古特大黄提取物不同成分泻下作用的比较研究[J].中国中药杂志,2007,32(2)：137.
[3]朱诗塔,李新中,雷鹏,等.不同煎煮法对大黄没食子酸含量的影响及与小肠推进作用的相关性[J].中南药学,2008,6(6)：714-717.
[4]黄建贤,陈少玲,黎惠娟,等.大黄促进小鼠小肠溶菌酶分泌的实验研究[J].中国医师杂志,2005,7(10)：1350-1351.
[5]宁康健,黄永梅,吕锦芳.大黄的不同处理对兔离体十二指肠张力的影响[J].中国中医药科技,2006,13(3)：173-174.
[6]李飞艳,邱赛红,尹健康,等.常用苦寒药对大鼠胃肠激素影响的研究[J].湖南中医药大学学报,2007,27(1)：9-11.
[7]张军.大黄对溃疡性结肠炎大鼠结肠电活动的影响[J].现代中西医结合杂志,2003,12(16)：1711.
[8]刘莉,梅其炳,周四元,等.大黄多糖对TNBS诱导大鼠结肠炎的治疗作用[J].中国中药杂志,2003,28(3)：246-249.
[9]刘海林.钙离子与肝细胞损伤及钙通道阻滞剂的保护作用[J].中华消化杂志,1993,13(2)：101.
[10]刘莉,梅其炳,李宝莉·唐古特大黄多糖对小鼠急性肝损伤的保护作用[J].第四军医大学学报,1999,20(6)：54.
[11]田萌,刘克武.大黄三种有效成分对胰蛋白酶活性及结构的影响[J].四川大学学报(自然 科学版),2010,47(1)：192-196.
[12]刘晓红,赵雩卿,钱家鸣.大黄对大鼠急性出血性胰腺炎的影响[J].中华消化杂志,2004,24(1)：14-17.
[13]李建生,王冬,方建,等.不同剂量大黄营元影响脑缺血大鼠血栓形成及血小板和凝血功能[J].中国临床康复,2005,9(21)：142-144.
[14]彭广能,周皓,徐在品,等.大黄注射液对犬血管搭桥术后血液流变学参数的影响[J].中国兽医学报,2009,29(8)：1072-1075.
[15]高雅萍,焦东海,王梅莉,等.大黄提取片对单纯性肥胖大鼠高胰岛素血症的作用[J].中国实验方剂学杂志,2002,8(6)：29-31.
[16]侯雁,于世家.大黄醇提物对糖尿病大鼠胰岛素敏感性及脂肪细胞因子影响的研究[J].实用糖尿病杂志,2006,2(3)：45-46.
[17]杜鹃,王琼书,贾汝汉.大黄对肾硬化大鼠肾小球细胞增殖和p27蛋白表达的影响[J].医药导报,2005,24(4)：278-281.
[18]崔春黎,马骏,肖自力,等.大黄对糖尿病肾病大鼠肾皮质基因表达谱的影响[J].中华肾脏病杂志,2002,18(4)：285-290.
[19]黄翠玲,李才.大黄对糖尿病大鼠肾皮质和尿前列腺素及血栓素水平的影响[J].中国中医药信息杂志,2003,10(12)：23-24.
[20]鲍军强,李锋,张文生,等.大黄对大鼠肾脏水通道蛋白2、4表达的影响[J].中国中西医结合杂志,2008,28(12)：1108-1111.
[21]王晓华,王笑云,熊明霞,等.大黄蒽醌提取物缓解小鼠肾组织纤维化作用的实验研究[J].中华肾脏病杂志,2006,22(6)：360-363.
[22]夏育民,徐世正.大黄素对狼疮样BXSB小鼠的影响[J].华中医学杂志,2003,27(2)：63-64.
[23]纪泽泉,黄翠雯,梁成结,等.大黄酸对肾小球硬化肾皮质凋亡蛋白酶-3活性及细胞凋亡的影响[J].中华医学杂志,2005,85(26)：1837-1841.
[24]张杰,屈燧林,王剑勤.大黄素对血管紧张素Ⅱ刺激人肾成纤维细胞增殖、胶原表达的抑制效应研究[J].四川医学,2002,23(11)：1114-1116.
[25]刘志红,郑敬民,吴义超,等.db/db糖尿病肾病小鼠肾脏基因表达谱及大黄酸对其的影响[J].肾脏病与透析肾移植杂志,2002,11(3)：201-204.
[26]郭啸华,刘志红,彭艾,等.大黄酸对2型糖尿病肾病大鼠疗效观察[J].中国肾脏病杂志, 2002,18(4)：280-284.
[27]鄣啸华,刘志红,王建平,等.大黄酸对NOD小鼠糖尿病肾病的治疗作用观察[J].肾脏病与透析肾移植杂志,2002,11(1)：11-16.
[28]朱加明,刘志红,黄燕飞,等.大黄酸对db/db小鼠糖尿病肾病疗效的观察[J].肾脏病与透析肾移植杂志,2002,11(1)：3-10.
[29]谭正怀,沈映君,赵军宁,等.大黄酸对人肾小球系膜细胞功能的影响[J].药学学报,2004,39(11)：881-886.
[30]赵焕利,李建国,黄祖敏,等.大黄对多发性创伤兔血中细胞因子变化的影响[J].遵义医学院学报,2007,30(1)：25-27.
[31]孙丽霞,任金荣,单保恩,等。大黄制剂对小鼠的急性毒性和自然免疫调节作用[J].癌变·畸变·突变,2006,18(1)：35.
[32]苗明三,苗艳艳.大黄多糖对衰老模型小鼠胸腺、脾脏影响的病理观察[J].中华实用中西医杂,2004,4(17)：146-147.
[33]邓廷飞,梁李广.大黄抗衰老的实验研究[J].中华中医药学刊,2007,25(7)：1510.
[34]党双锁,张正国,袁利超,等.大黄素和黄芪多糖对大鼠实验性肝癌的抑制作用[J].西安交通大学学报,2006,27(3)：250-253.
[35]李慧,杨洁,康迅雷,等.大黄素提高砷剂抑瘤效果的体内研究[J].上海第二医科大学学报,2005,25(7)：666-670.
[36]姬志伟,罗连城,解基良,等.大承气冲剂和大黄的体外抑菌作用[J].中国中西医结合外科杂志,2003,9(6)：451-453.
[37]蔡晓剑,李松龄,张荣.大黄对辣椒疫霉菌的室内抑菌研究[J].北方园艺,2006,(5)：169.
[38]吴生满.大黄提取物对氧自由基(O2-·)的清除效果研究[J].青海大学学报(自然科学版),2006,24(2)：80-81.
[39]王棋.不同提取方式对大黄抗氧化作用的影响[J].包头医学,2004,28(4)：28-30.
[40]李建生,王至婉,侯秀娟,等.大黄保护老龄大鼠脑缺血损害及其对细胞因子的影响[J].中国临床康复,2004,8(4)：676-678.
[41]李琳,段光琳,赵丽,等.大黄素对去卵巢大鼠和经β淀粉样蛋白作用的培养神经元的保护作用[J].医药导报,2005,24(2)：100-103.
[42]李琳,段光琳,赵丽,等.大黄素对去卵巢鼠行为及受β淀粉蛋白作用的培养神经元的保护作用[J].中国神经科学杂志,2004,20(5)：342-346.
[43]沈丽霞,李淑娟,张丹参,等.大黄酚对小鼠记忆障碍的作用及其机制分析[J].中国药理学通报,2003,19(8)：906-908.
[44]武海霞,张丹参,张力,等.大黄酚对β-淀粉样蛋白25-35致小鼠学习记忆障碍的改善作用[J].中国老年学杂志,2009,29(16)：2052-2054.
[45]王树,张丹参,张力,等.大黄酚对脑缺血再灌注小鼠记忆功能的保护作用[J].中国老年学杂志,2009,29(15)：1934-1936.
[46]王薇,杨占秋,钟琼,等.大黄茶抗柯萨奇病毒B2(CVB3)的实验研究[J].中医药学报,2005,33(4)：17-18.
[47]尹刚,王志强,黄美蓉.大黄和肉苁蓉提取物对感染性休克模型大鼠肺损伤的影响[J].山东中医杂志,2003,22(11)：691-693.
[48]杨明华,武影,沈海雁.大黄蒽醌衍生物对人牙周膜细胞作用的体外实验研究[J].牙体牙髓牙周病学杂志,2002,12(7)：355-357.
[49]孙庆顺,刘亚梅,李维善,等.大黄素对牙周炎大鼠牙龈组织中IL-6的影响[J].黑龙江医药科学,2008,31(6)：39-40.
[50]李淑娟,沈丽霞,张丹参,等.大黄酚对小鼠缺氧及耐力的影响[J].张家口医学院学报,2002,19(6)：10-11.
[51]林胜璋,徐贤绸,童洪飞,等.大黄素对大鼠肝移植排斥反应的抑制作用[J].中华器官移植杂志,2007,28(3)：147-149.
[52]李志刚,王章阳,刘松青.大黄素对家兔离体主动脉平滑肌细胞增殖影响的可能途径[J].中国临床康复,2005,9(26)：134-135.
[53]潘浩,刘学波,张红旗,等.大黄素对人血管平滑肌细胞增殖的影响[J].解剖学研究,2004,26(2)：121-123.
[54]王翔飞,葛均波,孙爱军,等.大黄素通过p53途径抑制血管平滑肌细胞增殖的实验研究[J].中华心血管病杂志,2006,34(1)：44-49.
[55]何烨,马力扬,李志强,等.大黄酸对大鼠离体子宫平滑肌运动的影响[J].沈阳药科大学学报,2007,24(4)：242-248.
[56]张喜平,程琪辉,裘怡.大黄素胶囊小鼠急性毒性试验[J].医学研究杂志,2007,36(6)：47-50.
[57]张陆勇.大黄总蒽醌对SD大鼠灌胃给药的长期毒性研究[J].中国生化药物杂志,2004,25(4)：206-209
[58]王清秀,吴纯启,廖明阳.大黄及其主要成分的毒性毒理研究[J].毒理学杂志,2007,21(4)：301-302.
[59]National Toxicology Program,2001. Technical report on the toxicology and carcinogenesis studies of emodin in F344/N rats and B6C3F1 mice. Series No.493. NIH Publication No. 01-3952. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC.
[60]Yan M, Zhang LY, Sun LX, Jiang ZZ, Xiao XH. Nephrotoxicity study of total rhubarb anthraquinones on Sprague Dawley rats using DNA microarrays[J]. Journal of Ethnopharmacology,2006,107(2):308-311.
[61]Mueller SO, Stopper H., Dekant W. Biotransformation of the anthraquinones emodin and chrysophanol by cytochrome P450 enzymes:Bioactivation to genotoxic metabolites[J]. Drug Metabolism and Disposition,1998,26:540-546.
[62]Wang CF, Wu XD, Chen M, Duan WG, Sun LX, Yan M, Zhang LY. Emodin induces apoptosis through caspase 3-dependent pathway in HK-2 cells[J]. Toxicology,2007,231(2-3): 120-128
[2]刘树民,罗明媚,李玉洁.中药毒性理论及研究进展[J].世界科学技术-中药现代化,2003,5(3)：45-47.
[3]王峥涛.中药毒性本质的科学研究[J].中药药性理论继承与创新研究简报,2007,(1)：16-20.
[4]肖小河,金城,王伽伯,等.中医药学科发展报告[M].北京：中国科学技术文献出版社,2006.
[5]彭成,王昌恩,林娜.有毒中药毒效相关性基础研究的意义与实践[J].中药药理与临床,2008,24(1)：71-73.
[6]雷载权,张廷模.中华临床中药学[M].北京：人民卫生出版社,1998：143.
[7]常惟智.影响中药功效及作用方向选择性之相关因素探析[J].中医药学报,2005,33(2)：12-13.
[8]余葱葱,郭力,彭成.毒效双组分控制有毒中药附子临床单煎工艺研究[J].时珍国医国药,2009,20(12)：3087-3089.
[9]王嫣虹,彭成,朱力阳.大黄素对结肠Cajal间质细胞毒效作用的研究[J].中国中西医合消化杂志,2010,18(3)：141-144.
[10]阮雯聪.影响中药功效与毒副作用的因素[J].浙江中西医结合杂志,2008,18(3)：197-198.
[11]由凤鸣,贾波,邓中甲.从剂量对中药功效发挥方向的影响论中药的矢量性[J].辽宁中医药大学学报.2010,12(5)：120-121.
[12]National Toxicology Program,2001. Technical report on the toxicology and carcinogenesis studies of emodin in F344/N rats and B6C3FI mice. Series No.493. NIH Publication No. 01-3952. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC.
[13]王伽伯,马永刚,张萍.炮制对大黄化学成分和肝肾毒性的影响及其典型相关分析[J].药学学报,2009,44(8)：885-890.
[14]谭余庆.对大黄素及大黄蒽醌化合物肾毒性和致癌性的再认识.国家药品审评中心电子刊物.
[15]Wang CF, Wu XD, Chen M,. Emodin induces apoptosis through caspase 3-dependent pathway in HK-2 cells[J]. Toxicology,2007,231(2-3):120-128.
[16]Yan M, Zhang LY, Sun LX, Jiang ZZ, Xiao XH. Nephrotoxicity study of total rhubarb anthraquinones on Sprague Dawley rats using DNA microarrays[J]. Journal of Ethnopharmacology,2006,107(2):308-311.
[17]张陆勇,江振洲,蒲存海.大黄总蒽醌对SD大鼠灌胃给药的长期毒性研究[J].中国生化药物杂志,2004,25(4)：206-209.
[18]Mueller SO, Stopper H., Dekant W. Biotransformation of the anthraquinones emodin and chrysophanol by cytochrome P450 enzymes:Bioactivation to genotoxic metabolites [J]. Drug Metabolism and Disposition,1998,26:540-546.
[19]Xiaomei Li, Haiyan Wang. Chinese herbal medicine in the treatment of chronic kidney disease [J]. Advances in Chronic Kidney Disease,2005,12(3):276-281.
[20]肖炜,邓虹珠,马云.大黄治疗慢性肾功能衰竭的临床与实验研究概述[J].中国中药杂志,2002,27(4)：241-262.
[21]Mitsuma, T., Terasawa, K., Yokozawa, T., Oura, H. Rhubarb therapy in patients with chronic renal failure (Part 1) [J]. Journal of Medical and Pharmaceutical Society for WAKAN-YAKU,1984,1:266-278.
[22]Mitsuma, T., Yokozawa, T., Oura, H., Terasawa, K. Rhubarb therapy in patients with chronic renal failure (Part 2)[J]. Japanese Journal of Nephrology,1987,29:195-207.
[23]李孜,卿平,冀玲.大黄治疗慢性肾功能衰竭的系统评价[J].中国循证医学杂志,2004,4(7)：468-473.
[24]Leishi Li. End-stage renal disease in China[J]. Kidney International,1996,49:287-301.
[25]肖相如.大黄在慢性肾衰中的正确运用[J].辽宁中医杂志,2002,29(6)：311-312.
[26]Jiabo Wang, Yanling Zhao, Xiaohe Xiao. Assessment of the renal protection and hepatotoxicity of rhubarb extract in rats[J]. Journal of Ethnopharmacology.124 (2009) 18-25.
[27]周本宏,王慧媛,郭志磊.石榴皮鞣质对羟自由基和超氧阴离子自由基的清除作用[J].国医院药学杂志,2008,28(17)：1442-1445.
[28]明玉,倪为为.大黄中鞣质成分的分离与液相色谱/质谱联用分析[J].色谱,2004, 22(6)：605-608.
[29]焦西英,杨浩,钱新宏.EGCG对过氧化氢所致神经元过氧化损伤的保护作用[J].解剖学研究,2008,30(6)：401-404.
[30]Pamela Pedrielli, Laura M. Holkeri, Leif H. Skibsted.Antioxidant activity of (+)-catechin. Rate constant for hydrogen-atom transfer to peroxyl radicals [J]. Eur Food Res Technol (2001)213:405-408.
[31]Yohei Miyamoto, Mariko Sano. Johnl.Haylor. (EGCG)-induced apoptosis in normal rat kidney interstitial fibroblast(NRK-49F)cells [J]. Eur Food Res Technol (2001) 213: 405-408.
[32]邓勇,黄珀,陈同良.肾小管细胞氧化性损伤模型的建立[J].现代泌尿外科杂志,2004,9(3)：144-145
[33]叶俊生.左卡尼汀在肾小管上皮细胞氧化应激损伤中的保护作用研究[D].南方医科大学博士论文,2010.
[34]叶任高,陆再英.内科学第五版[M].北京：人民卫生出版社,2001：569.
[35]万德光,彭成,赵军宁.四川道地中药材志[M].成都：四川科学技术出版社,2005：94.
[36]孙玉琦,肖小河,马永刚.大黄煎煮过程中蒽醌类成分动态变化规律研究[J].解放军药学学报,2006,22：281-283.
[37]杨洋,张蔚,黄丽琼.槲皮素联合顺铂对宫颈癌HeLa细胞增殖及凋亡的影响[J].武汉大学学报(医学版),2009,30(5)：334-336.
[38]邓生琼,周显琎,王瑶.硫化氢对氧化损伤心肌细胞凋亡的影响[J].武汉大学学报(医学版).2009,30(1)：61-65.
[39]李瑞萍,胡雪梅,吕银凤.苦参碱对人卵巢癌细胞增殖的影响及其机制[J].西安交通大学学报(医学版).2010,31(5)：621-624.
[40]孙沂,樊宏伟,王书奎.氯吡格雷对人胃黏膜上皮细胞株GES-1增殖作用的影响[J].世华人消化杂志,2010,18(4)：329-334.
[41]胡良平.现代统计学与SAS应用[M].北京：军事医学科学出版社,2000：367.
[42]刘珺,徐选福,杨文娟.红花注射液对肝星状细胞HSC-T6增殖、凋亡凋亡相关基因表达的影响[M].中草药,2009,40(8)：1270-1274.
[43]Parikh CR, Edelstein CL, Devarajan P. Biomarkers of acute kidney injury:early diagnosis, Pathogenesis, and recovery [J]. J Investig Med.2007,55(7):333-40.
[44]Djamali, A. Oxidative stress as a common Pathway to chronic tubulointerstitial injury in kidney allografts [J]. Am. J. Physiol. Renal Physiol.293:445-455; 2007.
[45]曹纯章,杨绍娟,卜丽莎.过氧化氢诱导诱导心肌细胞凋亡性和非凋亡性死亡的研究[J].中国病理生理杂志,2000,16(5)：457-461.
[46]Thannickal VJ, Fanburg BL. Reactive oxygen species in cell signaling [J]. Am J Physiol Lung Cell Mol Physiol,2000,279(6):L1005-L1028.
[47]McCord JM:Human disease, free radicals, and the oxidant/antioxidant balance [J]. Clin Biochem,1993,26(5):3511-3517.
[48]Vaziri ND, Oveisi F, Ding Y. Role of increased oxygen free radical activity in the Pathogenesis of uremic hypertension [J]. Kidney Int.1998,53:1748-1754.
[49]TePel M, Echelmeyer M, Orie NN,et al. Increased intracellular reactive oxygen Species in Patients with end-stage chronic renal failure:effect of hemodialysis[J].Kidney Int.2000,58:867-872.
[50]Parthasarathy S, Santanam N, Ramachandran S,etal.Oxidants and antioxidant in atherogenesis:an appraisal [J]. Lipid Res.1999,40:2143-2157.
[51]Baliga R, Ueda N, Walker PD, et al. Oxidant mechanisms in toxic renal failure [J]. Drug Metab Rev,1999,31(4):971-997.
[52]Valko,M., Leibfritz,D., Moncol, J.et al. Free radicals and antioxidants in normal physiological functions and human disease [J]. Int.J.Biochem. Cell.Biol.39:44-84;2007.
[53]崔瑛,许伟,海霞.试论中药毒的性能特征[J].河南中医,1997,17(5)：313-315.
[54]张廷模.临床中药学[M].北京：中国中医药出版社,2004,63.
[55]杨藻宸.医学药理学[M].北京：人民卫生出版社,1994：48.
[56]万德光.中药品种品质与药效[M].上海：上海科学技术出版社,2006：94.
[57]王筠默.中药药理学[M].上海：上海科学技术出版社,1985：12.
[58]李萌,王岩.乐陵大枣与静海金丝小枣药理作用的比较[J].中药材,1993,16(6)：35.
[59]聂淑琴,李兰芳,杨庆.5种产地苍术提取物主要药理作用比较研究[J].中国实验方剂学杂志,2002(81)：358-363.
[60]阴健,郭力弓.中药现代研究与临床应用[M].北京：学苑出版社,1993：4,6,24,596.
[61]徐楚江.中药炮制学[M].上海：上海科学技术出版社,1985：84.
[62]李仪奎,姜名瑛.中药药理学[M].北京：中国中医药山版社,1992：17.
[63]Yi Qing, Jia-bo Wang, Wei-jun Kong, et al. The diarrhoeogenic and antidiarrhoeal bidirectional effects of rhubarb and its potential mechanism [J]. Journal of Ethnopharmacology 133 (2011) 1096-1102.
[64]赵红玉,周春祥.基于药、毒效成分辩证关系探讨有毒中药研究思路[J].中草药,2008,39(11)：1601-1603.
[65]彭成.中医药动物实验方法学[M].北京：人民卫生出版社,2008：1223.
[66]王志鹏,梅其炳,刘琳娜.庆大霉素对人肾小管上皮细胞系(HK-2)热休克蛋白(HSP)70表达的影响[J].中国药理学通报,2006,22(4)：497-500.
[67]周桂凤,刘栋,乔玉峰.铅对体外培养HK-2细胞毒性及碘化钾的拮抗作用[J].毒理学杂志,2008,22(4)：283-286.
[68]唐功耀,田雪飞.马兜铃酸对人肾细胞作用的实验研究[J].中华肾脏病杂志,2002,18(4)：266-269.
[69]李彪,李晓玫,张翠英.马兜铃内酞胺对肾小管上皮细胞的损伤作用[J],中国中药杂志,2004：28(1)：79-83.
[70]王清秀.大黄及其主要成份的毒性毒理研究.军事医学科学院博士学位论文[D].2007.
[71]王清秀,吴纯启,杨红莲.大黄素对HK-2细胞周期增殖的影响毒理学杂志[J].2007,21(6)：440-444.
[72]李瑞萍,胡雪梅,吕银凤.苦参碱对人卵巢癌细胞增殖的影响及其机制[J].西安交通大学学报,2010,31(5)：621-623.
[73]卢薇娜.辛伐他汀和血脂康对糖尿病大鼠肾脏细胞凋亡的影响及其可能机制[D].2010.
[74]彭黎明,王曾礼.细胞凋亡的基础与临床[M].北京：人民卫生出版社,2000：2.
[75]王彩芳.鞣质生理活性研究进展[J].国外医学中医中药分册,2001,23(5)：278-282.
[76]Yoshioka H, Akai G, Yoshinage K. Protecting effect of a green tea percolate and its main constituents against gamma ray-induced scission of DNA[J]. Biosci Biotech Biochem, 1996,60:117-119.
[77]Yang GY, Liao J, Kim K, etal. Inhibition of growth and induction of apoptosis in human caneer cell lines by tea polyphenols[J]. Carcinogenesis,1998:19:611-616.
[78]Liang YC, Shoei L S. Lin JK, et al. Inhibition of cyclin-dependent kinases 2 and 4 activities as well as induction of cdk inhibitors P21 and P27 during growth arrest of human breast carcinoma cells by(-)-epigallocatechin-3-gallate[J]. J Cellular biochem.1999,75:1-8.
[79]Yohei MAYAMOTO, John.L HAYLOR, A.Meguid EL NAHAS. Cellular toxicity of catechin analogues containing gallte in opossum kidney Proximal tubular(OK) cells[J]. The Journal of toxicological sciences,29(1):47-52,2004.
[1]万德光.中药品种品质与药效[M].上海：上海科学技术出版社,2007：164-165.
[2]李文渊,童丽,等.唐古特大黄提取物不同成分泻下作用的比较研究[J].中国中药杂志,2007,32(2)：137.
[3]朱诗塔,李新中,雷鹏,等.不同煎煮法对大黄没食子酸含量的影响及与小肠推进作用的相关性[J].中南药学,2008,6(6)：714-717.
[4]黄建贤,陈少玲,黎惠娟,等.大黄促进小鼠小肠溶菌酶分泌的实验研究[J].中国医师杂志,2005,7(10)：1350-1351.
[5]宁康健,黄永梅,吕锦芳.大黄的不同处理对兔离体十二指肠张力的影响[J].中国中医药科技,2006,13(3)：173-174.
[6]李飞艳,邱赛红,尹健康,等.常用苦寒药对大鼠胃肠激素影响的研究[J].湖南中医药大学学报,2007,27(1)：9-11.
[7]张军.大黄对溃疡性结肠炎大鼠结肠电活动的影响[J].现代中西医结合杂志,2003,12(16)：1711.
[8]刘莉,梅其炳,周四元,等.大黄多糖对TNBS诱导大鼠结肠炎的治疗作用[J].中国中药杂志,2003,28(3)：246-249.
[9]刘海林.钙离子与肝细胞损伤及钙通道阻滞剂的保护作用[J].中华消化杂志,1993,13(2)：101.
[10]刘莉,梅其炳,李宝莉·唐古特大黄多糖对小鼠急性肝损伤的保护作用[J].第四军医大学学报,1999,20(6)：54.
[11]田萌,刘克武.大黄三种有效成分对胰蛋白酶活性及结构的影响[J].四川大学学报(自然 科学版),2010,47(1)：192-196.
[12]刘晓红,赵雩卿,钱家鸣.大黄对大鼠急性出血性胰腺炎的影响[J].中华消化杂志,2004,24(1)：14-17.
[13]李建生,王冬,方建,等.不同剂量大黄营元影响脑缺血大鼠血栓形成及血小板和凝血功能[J].中国临床康复,2005,9(21)：142-144.
[14]彭广能,周皓,徐在品,等.大黄注射液对犬血管搭桥术后血液流变学参数的影响[J].中国兽医学报,2009,29(8)：1072-1075.
[15]高雅萍,焦东海,王梅莉,等.大黄提取片对单纯性肥胖大鼠高胰岛素血症的作用[J].中国实验方剂学杂志,2002,8(6)：29-31.
[16]侯雁,于世家.大黄醇提物对糖尿病大鼠胰岛素敏感性及脂肪细胞因子影响的研究[J].实用糖尿病杂志,2006,2(3)：45-46.
[17]杜鹃,王琼书,贾汝汉.大黄对肾硬化大鼠肾小球细胞增殖和p27蛋白表达的影响[J].医药导报,2005,24(4)：278-281.
[18]崔春黎,马骏,肖自力,等.大黄对糖尿病肾病大鼠肾皮质基因表达谱的影响[J].中华肾脏病杂志,2002,18(4)：285-290.
[19]黄翠玲,李才.大黄对糖尿病大鼠肾皮质和尿前列腺素及血栓素水平的影响[J].中国中医药信息杂志,2003,10(12)：23-24.
[20]鲍军强,李锋,张文生,等.大黄对大鼠肾脏水通道蛋白2、4表达的影响[J].中国中西医结合杂志,2008,28(12)：1108-1111.
[21]王晓华,王笑云,熊明霞,等.大黄蒽醌提取物缓解小鼠肾组织纤维化作用的实验研究[J].中华肾脏病杂志,2006,22(6)：360-363.
[22]夏育民,徐世正.大黄素对狼疮样BXSB小鼠的影响[J].华中医学杂志,2003,27(2)：63-64.
[23]纪泽泉,黄翠雯,梁成结,等.大黄酸对肾小球硬化肾皮质凋亡蛋白酶-3活性及细胞凋亡的影响[J].中华医学杂志,2005,85(26)：1837-1841.
[24]张杰,屈燧林,王剑勤.大黄素对血管紧张素Ⅱ刺激人肾成纤维细胞增殖、胶原表达的抑制效应研究[J].四川医学,2002,23(11)：1114-1116.
[25]刘志红,郑敬民,吴义超,等.db/db糖尿病肾病小鼠肾脏基因表达谱及大黄酸对其的影响[J].肾脏病与透析肾移植杂志,2002,11(3)：201-204.
[26]郭啸华,刘志红,彭艾,等.大黄酸对2型糖尿病肾病大鼠疗效观察[J].中国肾脏病杂志, 2002,18(4)：280-284.
[27]鄣啸华,刘志红,王建平,等.大黄酸对NOD小鼠糖尿病肾病的治疗作用观察[J].肾脏病与透析肾移植杂志,2002,11(1)：11-16.
[28]朱加明,刘志红,黄燕飞,等.大黄酸对db/db小鼠糖尿病肾病疗效的观察[J].肾脏病与透析肾移植杂志,2002,11(1)：3-10.
[29]谭正怀,沈映君,赵军宁,等.大黄酸对人肾小球系膜细胞功能的影响[J].药学学报,2004,39(11)：881-886.
[30]赵焕利,李建国,黄祖敏,等.大黄对多发性创伤兔血中细胞因子变化的影响[J].遵义医学院学报,2007,30(1)：25-27.
[31]孙丽霞,任金荣,单保恩,等。大黄制剂对小鼠的急性毒性和自然免疫调节作用[J].癌变·畸变·突变,2006,18(1)：35.
[32]苗明三,苗艳艳.大黄多糖对衰老模型小鼠胸腺、脾脏影响的病理观察[J].中华实用中西医杂,2004,4(17)：146-147.
[33]邓廷飞,梁李广.大黄抗衰老的实验研究[J].中华中医药学刊,2007,25(7)：1510.
[34]党双锁,张正国,袁利超,等.大黄素和黄芪多糖对大鼠实验性肝癌的抑制作用[J].西安交通大学学报,2006,27(3)：250-253.
[35]李慧,杨洁,康迅雷,等.大黄素提高砷剂抑瘤效果的体内研究[J].上海第二医科大学学报,2005,25(7)：666-670.
[36]姬志伟,罗连城,解基良,等.大承气冲剂和大黄的体外抑菌作用[J].中国中西医结合外科杂志,2003,9(6)：451-453.
[37]蔡晓剑,李松龄,张荣.大黄对辣椒疫霉菌的室内抑菌研究[J].北方园艺,2006,(5)：169.
[38]吴生满.大黄提取物对氧自由基(O2-·)的清除效果研究[J].青海大学学报(自然科学版),2006,24(2)：80-81.
[39]王棋.不同提取方式对大黄抗氧化作用的影响[J].包头医学,2004,28(4)：28-30.
[40]李建生,王至婉,侯秀娟,等.大黄保护老龄大鼠脑缺血损害及其对细胞因子的影响[J].中国临床康复,2004,8(4)：676-678.
[41]李琳,段光琳,赵丽,等.大黄素对去卵巢大鼠和经β淀粉样蛋白作用的培养神经元的保护作用[J].医药导报,2005,24(2)：100-103.
[42]李琳,段光琳,赵丽,等.大黄素对去卵巢鼠行为及受β淀粉蛋白作用的培养神经元的保护作用[J].中国神经科学杂志,2004,20(5)：342-346.
[43]沈丽霞,李淑娟,张丹参,等.大黄酚对小鼠记忆障碍的作用及其机制分析[J].中国药理学通报,2003,19(8)：906-908.
[44]武海霞,张丹参,张力,等.大黄酚对β-淀粉样蛋白25-35致小鼠学习记忆障碍的改善作用[J].中国老年学杂志,2009,29(16)：2052-2054.
[45]王树,张丹参,张力,等.大黄酚对脑缺血再灌注小鼠记忆功能的保护作用[J].中国老年学杂志,2009,29(15)：1934-1936.
[46]王薇,杨占秋,钟琼,等.大黄茶抗柯萨奇病毒B2(CVB3)的实验研究[J].中医药学报,2005,33(4)：17-18.
[47]尹刚,王志强,黄美蓉.大黄和肉苁蓉提取物对感染性休克模型大鼠肺损伤的影响[J].山东中医杂志,2003,22(11)：691-693.
[48]杨明华,武影,沈海雁.大黄蒽醌衍生物对人牙周膜细胞作用的体外实验研究[J].牙体牙髓牙周病学杂志,2002,12(7)：355-357.
[49]孙庆顺,刘亚梅,李维善,等.大黄素对牙周炎大鼠牙龈组织中IL-6的影响[J].黑龙江医药科学,2008,31(6)：39-40.
[50]李淑娟,沈丽霞,张丹参,等.大黄酚对小鼠缺氧及耐力的影响[J].张家口医学院学报,2002,19(6)：10-11.
[51]林胜璋,徐贤绸,童洪飞,等.大黄素对大鼠肝移植排斥反应的抑制作用[J].中华器官移植杂志,2007,28(3)：147-149.
[52]李志刚,王章阳,刘松青.大黄素对家兔离体主动脉平滑肌细胞增殖影响的可能途径[J].中国临床康复,2005,9(26)：134-135.
[53]潘浩,刘学波,张红旗,等.大黄素对人血管平滑肌细胞增殖的影响[J].解剖学研究,2004,26(2)：121-123.
[54]王翔飞,葛均波,孙爱军,等.大黄素通过p53途径抑制血管平滑肌细胞增殖的实验研究[J].中华心血管病杂志,2006,34(1)：44-49.
[55]何烨,马力扬,李志强,等.大黄酸对大鼠离体子宫平滑肌运动的影响[J].沈阳药科大学学报,2007,24(4)：242-248.
[56]张喜平,程琪辉,裘怡.大黄素胶囊小鼠急性毒性试验[J].医学研究杂志,2007,36(6)：47-50.
[57]张陆勇.大黄总蒽醌对SD大鼠灌胃给药的长期毒性研究[J].中国生化药物杂志,2004,25(4)：206-209
[58]王清秀,吴纯启,廖明阳.大黄及其主要成分的毒性毒理研究[J].毒理学杂志,2007,21(4)：301-302.
[59]National Toxicology Program,2001. Technical report on the toxicology and carcinogenesis studies of emodin in F344/N rats and B6C3F1 mice. Series No.493. NIH Publication No. 01-3952. U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC.
[60]Yan M, Zhang LY, Sun LX, Jiang ZZ, Xiao XH. Nephrotoxicity study of total rhubarb anthraquinones on Sprague Dawley rats using DNA microarrays[J]. Journal of Ethnopharmacology,2006,107(2):308-311.
[61]Mueller SO, Stopper H., Dekant W. Biotransformation of the anthraquinones emodin and chrysophanol by cytochrome P450 enzymes:Bioactivation to genotoxic metabolites[J]. Drug Metabolism and Disposition,1998,26:540-546.
[62]Wang CF, Wu XD, Chen M, Duan WG, Sun LX, Yan M, Zhang LY. Emodin induces apoptosis through caspase 3-dependent pathway in HK-2 cells[J]. Toxicology,2007,231(2-3): 120-128