六神丸急性毒性及其抗肿瘤作用研究
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摘要
目的：六神丸因含雄黄,其安全性受到质疑。本文旨在探讨六神丸及其组分雄黄、蟾酥与无机砷急性毒性差异。方法：小鼠一次灌胃给予六神丸、雄黄、蟾酥、砷酸氢二钠(As5+),8h后检测肝、肾、心病理,砷蓄积及金属毒性敏感基因(MT)的表达。结果：六神丸、雄黄、蟾酥对血谷丙转氨酶(ALT)和血尿素氮(UREA)及肝、肾、心组织病理,金属硫蛋白的表达无明显影响。而As5+明显升高血清中ALT、UREA水平,组织砷蓄积量远大于六神丸和雄黄,引起严重的病理改变及诱导MT表达。结论：六神丸急性毒性远小于As5+,也不等同于雄黄和蟾酥急性毒性的简单叠加,用总含砷量去评价六神丸的毒性是不科学的。
目的探讨六神丸(Liu-Shen-Wan, LSW)的体外抗肿瘤作用及其作用机制。方法体外培养人肝癌Hep3B细胞,给予不同浓度的LSW及其他含砷化合物,MTS法检测细胞增殖；原子吸收法测定细胞内砷含量；流式细胞术检测细胞周期和细胞凋亡；PCR测定BAD基因mRNA水平。结果与顺铂及砷化物比较,小剂量LSW明显抑制Hep3B细胞增殖,可增加细胞内砷含量和细胞凋亡百分率；增加Hep3B细胞BAD基因mRNA表达。结论LSW对人肝癌Hep3B细胞具有抑制增值和杀灭的作用,机制涉及细胞毒作用、上调诱导细胞凋亡相关基因BAD的mRNA表达。
Aim To compare acute toxicity of Liu-Shen-Wan (LSW), realgar, Chan-Su, and sodium arsenate in mice. Method Mice were given orally LSW, realgar, arsenate (As5+), and Chan-Su. Acute toxicity and tissue As accumulation were determined 8 hrs later.
Results As5+ increased alanine aminotransferase (ALT) and blood urea nitrogen (UREA) levels, indicative of liver and kidney injury. No elevation of ALT and UREA was observed in LSW, realgar and Chan-Su groups. Histopathology showed more lesions in As5+ -treated liver, kidney and heart, while these lesions were mild or absent in Chan-Su, realgar and LSW groups,In Chan-Su-treated heart, cardiac lesions were observed, but were less severe as compared to As5+. As5+ administration resulted in As accumulation in liver (3560 ng·g-1), kidney (4570 ng·g-1) and heart (410 ng·g-1), much higher than that after LSW (260,180 and 160 ng·g-1). Metallothionein expression was dramatically induced by As5+ in liver (14x), kindey (25x), and heart (180x), but was basically unchanged after LWS, realgar and Chan-Su. Conclusions LSW and realgar are much less toxic acutely than As5+, and the use of total As content to evaluate LSW safety appear to be inappropriate.
Objective To examine the effect of Liu-Shen-Wan (LSW) on the proliferation of the human hepatocellular carcinoma Hep3B cells and the potential mechanisms. Methods The proliferative effects of Hep3B cells were determined by the MTS assay. The cell cycle was examined by flow cytometry. The expression of related genes was detected by real-time RT-PCR. Arsenic (As) concentration in cells was examined by Atomic Absorption Spectrometry. Results The proliferation of Hep3B cells for 48 h was suppressed significantly by LSW and NaAs2O3. Cellular As contents in LSW group were higher than other group treated with AS4S4, but were much lower than As3+, As5+ and ATO. LSW induced apoptosis of the cells as evidenced by flow cytometry,and increased expression of BAD. Conclusion LSW was effective in the growth inhibition of the human hepatic carcinoma Hep3B cells. The mechanisms appeared to be related to increased cellular As contents and increased tumor cell apoptosis by regulating gene expressions.
目的探讨六神丸(Liu-Shen-Wan, LSW)的体外抗肿瘤作用及其作用机制。方法体外培养人肝癌Hep3B细胞,给予不同浓度的LSW及其他含砷化合物,MTS法检测细胞增殖；原子吸收法测定细胞内砷含量；流式细胞术检测细胞周期和细胞凋亡；PCR测定BAD基因mRNA水平。结果与顺铂及砷化物比较,小剂量LSW明显抑制Hep3B细胞增殖,可增加细胞内砷含量和细胞凋亡百分率；增加Hep3B细胞BAD基因mRNA表达。结论LSW对人肝癌Hep3B细胞具有抑制增值和杀灭的作用,机制涉及细胞毒作用、上调诱导细胞凋亡相关基因BAD的mRNA表达。
Aim To compare acute toxicity of Liu-Shen-Wan (LSW), realgar, Chan-Su, and sodium arsenate in mice. Method Mice were given orally LSW, realgar, arsenate (As5+), and Chan-Su. Acute toxicity and tissue As accumulation were determined 8 hrs later.
Results As5+ increased alanine aminotransferase (ALT) and blood urea nitrogen (UREA) levels, indicative of liver and kidney injury. No elevation of ALT and UREA was observed in LSW, realgar and Chan-Su groups. Histopathology showed more lesions in As5+ -treated liver, kidney and heart, while these lesions were mild or absent in Chan-Su, realgar and LSW groups,In Chan-Su-treated heart, cardiac lesions were observed, but were less severe as compared to As5+. As5+ administration resulted in As accumulation in liver (3560 ng·g-1), kidney (4570 ng·g-1) and heart (410 ng·g-1), much higher than that after LSW (260,180 and 160 ng·g-1). Metallothionein expression was dramatically induced by As5+ in liver (14x), kindey (25x), and heart (180x), but was basically unchanged after LWS, realgar and Chan-Su. Conclusions LSW and realgar are much less toxic acutely than As5+, and the use of total As content to evaluate LSW safety appear to be inappropriate.
Objective To examine the effect of Liu-Shen-Wan (LSW) on the proliferation of the human hepatocellular carcinoma Hep3B cells and the potential mechanisms. Methods The proliferative effects of Hep3B cells were determined by the MTS assay. The cell cycle was examined by flow cytometry. The expression of related genes was detected by real-time RT-PCR. Arsenic (As) concentration in cells was examined by Atomic Absorption Spectrometry. Results The proliferation of Hep3B cells for 48 h was suppressed significantly by LSW and NaAs2O3. Cellular As contents in LSW group were higher than other group treated with AS4S4, but were much lower than As3+, As5+ and ATO. LSW induced apoptosis of the cells as evidenced by flow cytometry,and increased expression of BAD. Conclusion LSW was effective in the growth inhibition of the human hepatic carcinoma Hep3B cells. The mechanisms appeared to be related to increased cellular As contents and increased tumor cell apoptosis by regulating gene expressions.
引文
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