CYP1A2 Depletion by shRNA Enhances Hepatotoxicity of Extract of Polygoni Multiflori Radix and Related Monomer Compositions

doi:10.19803/j.1672-8629.2021.03.03

Abstract

Abstract: Objective To construct the CYP1A2 depletion human liver cell lines by shRNA, and to explore the relationship between depletion of CYP1A2 and hepatotoxicity of extract of Polygoni Multiflori Radix (PMR)and related monomer compositions. Method We constructed stably transfected L02-shCYP1A2 and HepaRG-shCYP1A2 cells with depletion of CYP1A2 by shRNA, verified by RT-qPCR. CYP1A2 depletion L02 and HepaRG cells were treated for 48 h with extract of PMR, and related monomer compositions included 2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-glucoside (TSG), emodin (EM), aloe-emodin (AE), emodin-8-O-β-D-glucopyranoside (EG), gallic acid (GA), physcion (PH), rhein (RH). Cell viability was detected by MTT assay. Result Successfully constructed two stably transfected cell lines, named L02-shCYP1A2 and HepaRG-shCYP1A2 cells, and the mRNA expression of CYP1A2 decreased by 59.81% in L02-shCYP1A2 and 66.60% in HepaRG-shCYP1A2, respectively. After L02-shCYP1A2 were treated with PMR for 48 h, cytotoxicity were significantly increased (P<0.01). After HepaRG-shCYP1A2 were treated with PMR for 48 h, cytotoxicity were significantly increased (P<0.05, P<0.01). The cytotoxicity of GA was significantly increased in L02-shCYP1A2 cells and HepaRG-shCYP1A2 cells (P<0.01, P<0.01). The cytotoxicity of AE was also significantly increased in HepaRG-shCYP1A2 cells (P<0.01). Conclusion The depletion of CYP1A2 significantly increased the hepatotoxicity of PMR, of which GA and AE were suggested to be the related monomer compositions.

Key words: polygoni multiflori radix, liver toxicity, RNA interference, CYP1A2, Gallic acid, Aloe-emodin

CLC Number:

R994.13

QUAN Zhengyang, LI Dengke, LI Yiqun, WANG Chengyu, ZHOU Ming, HU Yinghuan, SUN Zhenxiao. CYP1A2 Depletion by shRNA Enhances Hepatotoxicity of Extract of Polygoni Multiflori Radix and Related Monomer Compositions[J]. Chinese Journal of Pharmacovigilance, 2021, 18(3): 213-219.

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