朱红膏中汞在破损皮肤模型大鼠体内的蓄积及毒性
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摘要
目的:研究朱红膏给药4周后汞在大鼠肝脏、肾脏和脑中的蓄积情况以及对肝肾的毒性作用,为朱红膏临床安全使用提供数据参考。方法:将44只SD大鼠随机分为正常组、朱红膏低、中、高剂量(1. 875,37. 5,75 mg·kg-1)组,连续给药4周后检测大鼠尿液、血液、肝脏、肾脏和脑中的汞含量,测定血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT),天门冬氨酸氨基转移酶(aspartate aminotransferase,AST),尿素氮(serum urea nitrogen,BUN)和血肌酐(serum creatinine,SCr)的浓度,尿液中β2-微球蛋白(β2-mi SCroglobulin,β2-BMG)和N-乙酰-β-D葡萄糖苷酶(N-acetyl-beta-D glucosidase,NAG)的浓度,组织切片进行苏木素-伊红(HE)染色观察肝肾组织形态。结果:朱红膏各给药组的血汞、尿汞含量明显高于正常组(P <0. 05,P <0. 01),且呈剂量依赖性;朱红膏高剂量组血清ALT明显升高,朱红膏中、高剂量组血清SCr明显升高(P <0. 05,P <0. 01),朱红膏低剂量组的生化指标没有改变;朱红膏中、高剂量会破坏肝小叶的结构,破坏肝索单层细胞放射状排列,使肾小管管腔扩张,肾小球萎缩,甚至肾小球缺失,朱红膏低剂量组对肝脏的组织形态无显著影响,轻微改变肾脏组织形态。结论:朱红膏在低剂量下使用一个月基本安全,但长期大量使用会造成汞在肝脏、肾脏和脑蓄积,并导致肝肾毒性。讨论:未发现比肝肾实质性病变更灵敏的指标用于监控其肝肾毒性,但尿汞和血汞的含量升高有助于预知其毒性,可对汞的暴露量与毒性的关系做进一步的探讨。研究为朱红膏临床合理安全使用以及毒性的监控提供了一定的参考。
Objective: To study the accumulation of mercury in liver,kidney and brain of rats and its toxicity on liver and kidney after 4 weeks of administration of different doses of Zhuhong ointment,in order to provide data reference for the safe clinical use of Zhuhong ointment. Method: Forty-four Sprague-Dawley( SD)rats were randomly divided into 4 groups: control group,normal-dose group( 1. 875 mg·kg-1),medium-dose group( 37. 5 mg·kg-1),and high-dose group( 75 mg·kg-1). After transdermal administration for consecutive4 weeks,the mercury content in the urine,blood,liver,kidney and brain of the rats was measured. In addition,serum alanine aminotransferase( ALT),serum aspartate aminotransferase( AST),serum urea nitrogen( BUN),and serum creatinine( SCr),urine β2-mi SCroglobulin( β2-BMG) and urine N-acetyl-beta-D glucosidase( NAG)contents were measured, and pathological morphology changes of liver and kidney were observed. Result:Compared with the control group,the levels of blood mercury and urine mercury in Zhuhong ointment groups showed significant increases after administration for 4 weeks( P < 0. 05,P < 0. 01). The level of serum ALT was significantly elevated in the high-dose group,and serum SCr was significantly elevated in middle and high-dose groups( P < 0. 05,P < 0. 01). Biochemical index in the normal dose remained unchanged. Medium and high doses of Zhuhong ointment destroyed the structure of the hepatic lobule and the radial arrangement of the liver monolayer,shrank the lumen and glomerular,and even led to glomerular balloon enlargement( P < 0. 05,P < 0. 01). Normal dose of Zhuhong ointment had no significant effect on liver tissue morphology,but slightly changed kidney tissue morphology. Conclusion: Zhuhong ointment is not toxic at the normal dose,but long-term use can lead to the accumulation of mercury in liver,kidney and bra,which causes liver and kidney toxicity. This study did not find a more sensitive indicator of liver and kidney toxicity than liver and kidney pathology. However,because the rising levels of urinary mercury and blood mercury may predict toxicity,the relationship between mercury exposure and toxicity could be further studied. This study provides a reference for the rational use and toxicity monitoring of Zhuhong ointment.
Objective: To study the accumulation of mercury in liver,kidney and brain of rats and its toxicity on liver and kidney after 4 weeks of administration of different doses of Zhuhong ointment,in order to provide data reference for the safe clinical use of Zhuhong ointment. Method: Forty-four Sprague-Dawley( SD)rats were randomly divided into 4 groups: control group,normal-dose group( 1. 875 mg·kg-1),medium-dose group( 37. 5 mg·kg-1),and high-dose group( 75 mg·kg-1). After transdermal administration for consecutive4 weeks,the mercury content in the urine,blood,liver,kidney and brain of the rats was measured. In addition,serum alanine aminotransferase( ALT),serum aspartate aminotransferase( AST),serum urea nitrogen( BUN),and serum creatinine( SCr),urine β2-mi SCroglobulin( β2-BMG) and urine N-acetyl-beta-D glucosidase( NAG)contents were measured, and pathological morphology changes of liver and kidney were observed. Result:Compared with the control group,the levels of blood mercury and urine mercury in Zhuhong ointment groups showed significant increases after administration for 4 weeks( P < 0. 05,P < 0. 01). The level of serum ALT was significantly elevated in the high-dose group,and serum SCr was significantly elevated in middle and high-dose groups( P < 0. 05,P < 0. 01). Biochemical index in the normal dose remained unchanged. Medium and high doses of Zhuhong ointment destroyed the structure of the hepatic lobule and the radial arrangement of the liver monolayer,shrank the lumen and glomerular,and even led to glomerular balloon enlargement( P < 0. 05,P < 0. 01). Normal dose of Zhuhong ointment had no significant effect on liver tissue morphology,but slightly changed kidney tissue morphology. Conclusion: Zhuhong ointment is not toxic at the normal dose,but long-term use can lead to the accumulation of mercury in liver,kidney and bra,which causes liver and kidney toxicity. This study did not find a more sensitive indicator of liver and kidney toxicity than liver and kidney pathology. However,because the rising levels of urinary mercury and blood mercury may predict toxicity,the relationship between mercury exposure and toxicity could be further studied. This study provides a reference for the rational use and toxicity monitoring of Zhuhong ointment.
引文
[1]吕培文,张苍,宋孝瑜,等.朱红膏治疗慢性溃疡的临床研究[J].中国中西医结合外科杂志,2003,9(5):364-366.
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[3]王乐平,董建勋,罗玲,等.外用制剂朱红膏肾脏早期毒性敏感指标NAG、RBP的监测[J].中华中医药杂志,2011,26(10):2261-2264.
[4]林含,张旭辉,董建勋,等.朱红膏对破损皮肤模型大鼠汞蓄积及肾组织形态的影响[J].中国中药杂志,2012,37(6):739-743.
[5]孙新民,王旗,牟稷征,等.玉红膏单次给药家兔体内汞的吸收及毒性[J].中国实验方剂学杂志,2011,17(24):193-196.
[6]付中祥,杨虹,陈秀芬,等.朱砂、朱砂安神丸及氯化汞在小鼠体内吸收、分布对比[J].中国实验方剂学杂志,2013,19(1):162-166.
[7]谷颖敏,李咏梅,姜昕,等.朱砂灌胃给药对大鼠生育力与早期胚胎发育毒性的研究[J].中国实验方剂学杂志,2011,17(9):226-231.
[8]程金平,刘晓洁,冀秀玲,等.总汞、甲基汞和硒在汞暴露大鼠脑、肝、肾中的分布[J].中国环境科学,2006,26(3):376-379.
[9]黄继汉,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.
[10]食品中总汞及有机汞的测定:GB 5009. 17-2014[S].北京:中国标准出版社,2015.
[11] LU Y F,WU Q,LIANG S X,et al. Evaluation of hepatotoxicity potential of cinnabar-containing An-GongNiu-Huang Wan,a patent traditional Chinese medicine[J]. Regul Toxicol Pharmacol,2011, 60(2):206-211.
[12]王晓烨,林瑞超,田悦,等.朱红膏中汞在大鼠皮肤不同损伤状态下的体外透过速率比较研究[J].环球中医药,2018,11(6):823-826.
[13] Lohren H,Bornhorst J,Fitkau R,et al. Effects on and transfer a SCross the blood-brain barrier in vitrocomparison of organic and inorganic mercury species[J]. BMC Pharmacol Toxicol,2016,17(1):63.
[14]李均,付旭,姚兰,等.丹酚酸B,C分子药对配伍对UUO大鼠肾间质纤维化的保护作用及机制[J].中国实验方剂学杂志,2015,21(20):113-117.
[2]王桂英,林含,张旭辉,等.朱红膏治疗慢性皮肤溃疡的临床安全性研究[J].北京中医药,2016,35(1):70-72.
[3]王乐平,董建勋,罗玲,等.外用制剂朱红膏肾脏早期毒性敏感指标NAG、RBP的监测[J].中华中医药杂志,2011,26(10):2261-2264.
[4]林含,张旭辉,董建勋,等.朱红膏对破损皮肤模型大鼠汞蓄积及肾组织形态的影响[J].中国中药杂志,2012,37(6):739-743.
[5]孙新民,王旗,牟稷征,等.玉红膏单次给药家兔体内汞的吸收及毒性[J].中国实验方剂学杂志,2011,17(24):193-196.
[6]付中祥,杨虹,陈秀芬,等.朱砂、朱砂安神丸及氯化汞在小鼠体内吸收、分布对比[J].中国实验方剂学杂志,2013,19(1):162-166.
[7]谷颖敏,李咏梅,姜昕,等.朱砂灌胃给药对大鼠生育力与早期胚胎发育毒性的研究[J].中国实验方剂学杂志,2011,17(9):226-231.
[8]程金平,刘晓洁,冀秀玲,等.总汞、甲基汞和硒在汞暴露大鼠脑、肝、肾中的分布[J].中国环境科学,2006,26(3):376-379.
[9]黄继汉,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.
[10]食品中总汞及有机汞的测定:GB 5009. 17-2014[S].北京:中国标准出版社,2015.
[11] LU Y F,WU Q,LIANG S X,et al. Evaluation of hepatotoxicity potential of cinnabar-containing An-GongNiu-Huang Wan,a patent traditional Chinese medicine[J]. Regul Toxicol Pharmacol,2011, 60(2):206-211.
[12]王晓烨,林瑞超,田悦,等.朱红膏中汞在大鼠皮肤不同损伤状态下的体外透过速率比较研究[J].环球中医药,2018,11(6):823-826.
[13] Lohren H,Bornhorst J,Fitkau R,et al. Effects on and transfer a SCross the blood-brain barrier in vitrocomparison of organic and inorganic mercury species[J]. BMC Pharmacol Toxicol,2016,17(1):63.
[14]李均,付旭,姚兰,等.丹酚酸B,C分子药对配伍对UUO大鼠肾间质纤维化的保护作用及机制[J].中国实验方剂学杂志,2015,21(20):113-117.