雄黄微生物转化液RTS在新型牛黄解毒片中的减毒增效作用研究
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摘要
将雄黄微生物转化液RTS替代牛黄解毒片中的雄黄,制成新型牛黄解毒片,并研究新型牛黄解毒片的药效及毒性作用。采用嗜酸性氧化亚铁硫杆菌At. f BY3对雄黄进行生物浸出,制备雄黄微生物转化液RTS并测定砷含量;按照《中国药典》制备由RTS替代雄黄的新型牛黄解毒片进行后续研究;采用灌胃的方法研究新型牛黄解毒片的急性毒性和亚急性毒性,并采用对其解热、抗炎、镇痛药理作用进行对比研究; MTT测定两种复方药物对RAW264. 7细胞的增殖抑制作用和对TNF-α炎性因子的抑制作用。雄黄微生物转化液RTS可对牛黄解毒片中的雄黄进行替换,并使药物达到减毒增效的效果。
In this paper,the realgar in Chinese traditional formula Niuhuangjiedu tablets( POS) is replaced by realgar transforming solusion( RTS) to make a novel Niuhuangjiedu tablets( REP). Previous studies showed that RTS had a greater inhibition on tumor cells. In this study,the author aim to evaluate the toxicology and antipyretic,analgesic and anti-inflammatory effects of REP and its anti-inflammatory mechanism. RTS is extracted from realgar by bioleaching with the acidophilic thiobacillus ferrooxidans At. f BY3.Acute toxicity experiment and subacute toxicity experiment were used to evaluate toxicological effect of REP,and the antipyretic,analgesic and anti-inflammatory effect was studied by carrageenan heating experiment,hot plate experiment,and ear swelling experiment. MTT assay was used to determine the inhibitory effect of the two compounds on the proliferation of RAW264. 7 cells. Additionally,the inflammatory inhibition of the two compounds on TNF-α was measured. The results showed REP had no acute toxicity,LD50>12. 5 g/kg and HE staining of liver and kidney damage caused by REP were much lighter than POS. Heating experiment showed that temperature of rats decreased after treating with REP,meanwhile hot plate experiment showed that REP could slightly increase the pain threshold of mice and the degree of ear swelling could be decreased by REP treatment,which were similar to POS. MTT assay demonstrated that REP had a relative safety on RAW264. 7 cells than POS,which showed a concentration-dependent inhibition ratio.The expression of TNF-α decreased both by REP and POS. In conclusion,REP demonstrated no acute toxicity and had an equivalent antipyretic,analgesic and anti-inflammatory effect than POS. In addition,REP showed a less cytotoxicity on RAW264. 7 than POS with decreasing expression of TNF-α. All in all,RTS could have the effect of toxicity-attenuating and efficiency-enhancement when realgar was replaced in novel Niuhuangjiedu teblets.
In this paper,the realgar in Chinese traditional formula Niuhuangjiedu tablets( POS) is replaced by realgar transforming solusion( RTS) to make a novel Niuhuangjiedu tablets( REP). Previous studies showed that RTS had a greater inhibition on tumor cells. In this study,the author aim to evaluate the toxicology and antipyretic,analgesic and anti-inflammatory effects of REP and its anti-inflammatory mechanism. RTS is extracted from realgar by bioleaching with the acidophilic thiobacillus ferrooxidans At. f BY3.Acute toxicity experiment and subacute toxicity experiment were used to evaluate toxicological effect of REP,and the antipyretic,analgesic and anti-inflammatory effect was studied by carrageenan heating experiment,hot plate experiment,and ear swelling experiment. MTT assay was used to determine the inhibitory effect of the two compounds on the proliferation of RAW264. 7 cells. Additionally,the inflammatory inhibition of the two compounds on TNF-α was measured. The results showed REP had no acute toxicity,LD50>12. 5 g/kg and HE staining of liver and kidney damage caused by REP were much lighter than POS. Heating experiment showed that temperature of rats decreased after treating with REP,meanwhile hot plate experiment showed that REP could slightly increase the pain threshold of mice and the degree of ear swelling could be decreased by REP treatment,which were similar to POS. MTT assay demonstrated that REP had a relative safety on RAW264. 7 cells than POS,which showed a concentration-dependent inhibition ratio.The expression of TNF-α decreased both by REP and POS. In conclusion,REP demonstrated no acute toxicity and had an equivalent antipyretic,analgesic and anti-inflammatory effect than POS. In addition,REP showed a less cytotoxicity on RAW264. 7 than POS with decreasing expression of TNF-α. All in all,RTS could have the effect of toxicity-attenuating and efficiency-enhancement when realgar was replaced in novel Niuhuangjiedu teblets.
引文
[1]国家药典委员会.中华人民共和国药典[S].北京:中国医药科技出版社,2015.
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[5]马斌,李彤,姜泓.我国雄黄及其复方的毒副作用研究进展[J].中华中医药学刊,2013(8):1623-1625.
[6]Zhang X.Biological extraction of realgar by acidithiobacillus ferrooxidans and its in vitro and in vivo antitumor activities[J].Pharm Biol,2010,48(1):40-47.
[7]Liu DL.Realgar bioleaching solution is a less toxic arsenic agent in suppressing the Ras/MAPK pathway in Caenorhabditis elegans[J].Environm Toxicol Pharmacol,2013,35(2):292-299.
[8]Song P.Realgar transforming solution displays anticancer potential against human hepatocellular carcinoma Hep G2 cells by inducing ROS[J].Intern J Oncol,2017,50(2):660-670.
[9]Wang X.Autophagy enhanced antitumor effect in K562 and K562/ADM cells using realgar transforming solution[J].Biomed Pharmacother,2018,98:p.252-264.
[10]Wang X.Reversal effect of arsenic sensitivity in human leukemia cell line K562 and K562/ADM using realgar transforming solution[J].Biol Pharm Bull,2013,36(4):641-648.
[11]Saklatvala J,W Davis,F Guesdon.Interleukin 1(IL1)and tumour necrosis factor(TNF)signal transduction[J].Philos Trans R Soc Lond B Biol Sci,1996,351(1336):151-157.
[2]孟海琴,高淑华.牛黄解毒片的抗炎,抑菌作用研究[J].中国中药杂志,1992,17(12):747-749.
[3]张娟,汤毅珊,刘启德.雄黄中砷的毒代动力学及组织学分布研究[J].中国中药杂志,2010,35(13):1750-1753.
[4]苗加伟.牛黄解毒片、雄黄与其他砷化物急性毒性的比较研究[J].毒理学杂志,2011,25(3):217-221.
[5]马斌,李彤,姜泓.我国雄黄及其复方的毒副作用研究进展[J].中华中医药学刊,2013(8):1623-1625.
[6]Zhang X.Biological extraction of realgar by acidithiobacillus ferrooxidans and its in vitro and in vivo antitumor activities[J].Pharm Biol,2010,48(1):40-47.
[7]Liu DL.Realgar bioleaching solution is a less toxic arsenic agent in suppressing the Ras/MAPK pathway in Caenorhabditis elegans[J].Environm Toxicol Pharmacol,2013,35(2):292-299.
[8]Song P.Realgar transforming solution displays anticancer potential against human hepatocellular carcinoma Hep G2 cells by inducing ROS[J].Intern J Oncol,2017,50(2):660-670.
[9]Wang X.Autophagy enhanced antitumor effect in K562 and K562/ADM cells using realgar transforming solution[J].Biomed Pharmacother,2018,98:p.252-264.
[10]Wang X.Reversal effect of arsenic sensitivity in human leukemia cell line K562 and K562/ADM using realgar transforming solution[J].Biol Pharm Bull,2013,36(4):641-648.
[11]Saklatvala J,W Davis,F Guesdon.Interleukin 1(IL1)and tumour necrosis factor(TNF)signal transduction[J].Philos Trans R Soc Lond B Biol Sci,1996,351(1336):151-157.