佐太、β-HgS、HgCl_2对PC12细胞活性和凋亡相关基因表达影响的比较
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摘要
目的比较佐太、β-HgS、HgCl_2对PC12细胞活性和凋亡相关基因表达的影响。方法 MTT法测定3种药物对细胞活性的影响,实时荧光定量PCR法检测Bax、Bak、Bcl2、Fas、FasL表达。结果 0.250 g/L佐太作用2 h后细胞活性下降约10%,等汞量β-HgS使其下降12%左右,而含汞量只有佐太1/10的HgCl_2却导致其下降约70%;佐太降低Bax、Bak、Bcl-2、FasL表达,β-HgS降低Bax、Bak、Fas、FasL表达,HgCl_2降低Bcl-2表达而增加Fas、FasL表达。结论佐太、β-HgS对PC12细胞的毒性远小于HgCl_2,两者可能对细胞中线粒体凋亡通路的激活有抑制作用,而HgCl_2可能通过激活死亡受体通路诱导细胞凋亡。
引文
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[12] 朱洪梅, 魏立新, 杜玉枝, 等. 藏药佐太长期给药对小鼠毒性的初步研究[J]. 时珍国医国药, 2013, 24(8): 2022-2024.
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[2] 贾敏如, 卢晓琳, 马逾英. 初论我国少数民族使用矿物药的品种概况[J]. 中国中药杂志, 2015, 40(23): 4693-4702.
[3] 嘎玛曲培,土登格桑,格穷. 药用水银粉末(坐台)的加工方法:中国,CN1038406[P]. 1990-01-03.
[4] Zhao X Y, Sun M, Wang J X, et al. Characterization of tibetan medicine Zuota by multiple techniques[J]. Bioinorg Chem Appl, 2013, 2013: 198545.
[5] Li C, Yang H X, Du Y Z, et al. Chemical species, micromorphology, and XRD fingerprint analysis of Tibetan medicine Zuotai containing mercury[J]. Bioinorg Chem Appl, 2016, 2016: 7010519.
[6] 王东平, 魏立新, 杜玉枝, 等. 藏药“佐太”中硫化汞含量测定的方法学考察[J]. 时珍国医国药, 2010, 21(6): 1359-1361.
[7] 王维恩. 藏药“佐太”的粉末X射线衍射指纹图谱分析[J]. 中国中药杂志, 2014, 39(7): 1179-1184.
[8] 李岑, 占堆, 楞本才让, 等. 藏药佐太的化学成分、汞配位结构及微观形貌分析[J]. 光谱学与光谱分析, 2015, 35(4): 1072-1078.
[9] 赵静, 张明, 耿卢婧, 等. 藏药佐太在两种抑郁小鼠模型中的抗抑郁活性[J]. 中成药, 2016, 38(7): 1461-1467.
[10] 曾勇, 何毓敏, 刘颖, 等. 藏药“佐塔”对中枢神经系统的部分药理作用研究[J]. 四川中医, 2005, 23(11): 33-34.
[11] 何海洋,康峰,颜俊文, 等. 朱砂、朱砂安神丸与氯化汞、轻粉的急性毒性对比[J]. 中国实验方剂学杂志, 2011, 17(2): 219-223.
[12] 朱洪梅, 魏立新, 杜玉枝, 等. 藏药佐太长期给药对小鼠毒性的初步研究[J]. 时珍国医国药, 2013, 24(8): 2022-2024.
[13] Wu Q, Li W K, Zhou Z P, et al. The Tibetan medicine Zuotai differs from HgCl2 and MeHg in producing liver injury in mice[J]. Regul Toxicol Pharmacol, 2016, 78: 1-7.
[14] Shi J Z, Kang F, Wu Q, et al. Nephrotoxicity of mercuric chloride, methylmercury and cinnabar-containing Zhu-Sha-An-Shen-Wan in rats[J]. Toxicol Lett, 2011, 200(3): 194-200.
[15] Chuu J J, Liu S H, Lin-Shiau S Y. Differential neurotoxic effects of methylmercury and mercuric sulfide in rats[J]. Toxicol Lett, 2007, 169(2): 109-120.
[16] Liu J, Shi J Z, Yu L M, et al. Mercury in traditional medicines: is cinnabar toxicologically similar to common mercurials?[J]. Exp Biol Med (Maywood), 2008, 233(7): 810-817.
[17] Wu Q, Lu Y F, Shi J Z, et al. Chemical form of metals in traditional medicines underlines potential toxicity in cell cultures[J]. J Ethnopharmacol, 2011, 134(3): 839-843.
[18] 阎立峰. 藏药“佐太”的微结构与成分分析[J]. 中国藏学, 2007(3): 150-152, 174.
[19] Asharani P V, Low Kah Mun G, Hande M P, et al. Cytotoxicity and genotoxicity of silver nanoparticles in human cells[J]. ACS Nano, 2009, 3(2): 279-290.
[20] Hussain S M, Javorina A K, Schrand A M, et al. The interaction of manganese nanoparticles with PC-12 cells induces dopamine depletion[J]. Toxicol Sci, 2006, 92(2): 456-463.
[21] Gerlier D, Thomasset N. Use of MTT colorimetric assay to measure cell activation[J]. J Immunol Methods, 1986, 94(1-2): 57-63.
[22] Liu J, Lu Y F, Li W K, et al. Mercury sulfides are much less nephrotoxic than mercury chloride and methylmercury in mice[J]. Toxicol Lett, 2016, 262: 153-160.
[23] Dong W, Liu J, Wei L X, et al. Developmental toxicity from exposure to various forms of mercury compounds in medaka fish (Oryzias latipes) embryos[J]. Peer J, 2016, 4: e2282.
[24] Carranza-Rosales P, Said-Fernandez S, Sepulveda-Saavedra J, et al. Morphologic and functional alterations induced by low doses of mercuric chloride in the kidney OK cell line: ultrastructural evidence for an apoptotic mechanism of damage[J]. Toxicology, 2005, 210(2-3): 111-121.
[25] 孟祥东, 严云勤. BCL-2家族与线粒体对细胞凋亡的调控[J]. 细胞与分子免疫学杂志, 2005, 21(S1):S77-S79.
[26] Ormerod M G, O’neill C F, Robertson D, et al. Cisplatin induces apoptosis in a human ovarian carcinoma cell line without concomitant internucleosomal degradation of DNA[J]. Exp Cell Res, 1994, 211(2): 231-237.
[27] Kiefer M C, Brauer M J, Powers V C, et al. Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak[J]. Nature, 1995, 374(6524): 736-739.
[28] Munoz L E, van Bavel C, Franz S, et al. Apoptosis in the pathogenesis of systemic lupus erythematosus[J]. Lupus, 2008, 17(5): 371-375.
[29] Nagata S. Fas ligand-induced apoptosis[J]. Annu Rev Genet, 1999, 33:29-55.