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Safety evaluation of chinese medicine injections with a cell imaging-based multiparametric assay revealed a critical involvement of mitochondrial function in hepatotoxicity.

Wang M, Liu CX, Dong RR, He S, Liu TT, Zhao TC, Wang ZL, Shen XY, Zhang BL, Gao XM, Zhu Y - Evid Based Complement Alternat Med (2015)

Bottom Line: The safety of herbal medicine products has been a widespread concern due to their complex chemical nature and lack of proper evaluation methods.Our results suggested that the reported hepatotoxicity by one of the drugs, Fufangkushen injection, could be attributed at least in part to the interference of mitochondrial function in human HepG2 cells by some of its constituents.This method should be useful for both preclinical screen in a drug discovery program and postclinical evaluation of herbal medicine preparations.

View Article: PubMed Central - PubMed

Affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Nankai District, Tianjin 300193, China ; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tianjin 300457, China.

ABSTRACT
The safety of herbal medicine products has been a widespread concern due to their complex chemical nature and lack of proper evaluation methods. We have adapted a sensitive and reproducible multiparametric cell-based high-content analysis assay to evaluate the hepatic-safety of four Chinese medicine injections and validated it with classical animal-based toxicity assays. Our results suggested that the reported hepatotoxicity by one of the drugs, Fufangkushen injection, could be attributed at least in part to the interference of mitochondrial function in human HepG2 cells by some of its constituents. This method should be useful for both preclinical screen in a drug discovery program and postclinical evaluation of herbal medicine preparations.

No MeSH data available.


Related in: MedlinePlus

Representative images of the HCA analysis of HepG2 cells for the evaluation of nuclei number and morphology stained by 4 dyes in control (a, f, k, p, and u), Danhong injection (DHI) 100-fold dilution (b, g, l, q, and v), Xiangdan injection (XDI) 100-fold dilution (c, h, m, r, and w), Mailuoning injection (MLNI) 100-fold dilution (d, i, n, s, anad x), and Fufangkushen injection (FFKSI) 100-fold dilution (e, j, o, t, and y) groups. The cell number (CN) decreased by XDI 100-fold dilution and FFKSI 100-fold dilution (c, e). The intensity of Rhodamine 123 (green) was lower in FFKSI-treated group than control group indicating that the MMP decreased (o). Note: IC 50 of cell parameters associated with cell number (CN) was IC 501 and IC 50 of cell parameters associated with nuclear area (NA) was IC 502.
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fig3: Representative images of the HCA analysis of HepG2 cells for the evaluation of nuclei number and morphology stained by 4 dyes in control (a, f, k, p, and u), Danhong injection (DHI) 100-fold dilution (b, g, l, q, and v), Xiangdan injection (XDI) 100-fold dilution (c, h, m, r, and w), Mailuoning injection (MLNI) 100-fold dilution (d, i, n, s, anad x), and Fufangkushen injection (FFKSI) 100-fold dilution (e, j, o, t, and y) groups. The cell number (CN) decreased by XDI 100-fold dilution and FFKSI 100-fold dilution (c, e). The intensity of Rhodamine 123 (green) was lower in FFKSI-treated group than control group indicating that the MMP decreased (o). Note: IC 50 of cell parameters associated with cell number (CN) was IC 501 and IC 50 of cell parameters associated with nuclear area (NA) was IC 502.

Mentions: Five fixed concentrations (1000-, 800-, 500-, 300-, 100-fold dilution) of four injections, DHI, XDI, FFKSI, and MLNI were selected. As shown in Figure 3, the representative images were captured by the HCA method established for typical cytotoxic effects on hepatocyte induced by FFKSI 100-fold dilution, XDI 100-fold dilution, MLNI 100-fold dilution, and DHI 100-fold dilution. It is noteworthy that 1000-, 800-, 500-, 300- (P < 0.05), and 100-fold dilution (P < 0.01) of FFKSI significantly decreased the MMP by 11.1%–22.8% as compared to blank group (Figure 4(d)). XDI and FFKSI at 100-fold dilution caused a significant decrease of CN by 50.8% and 48.4% (P < 0.05), respectively (Figure 4(a)). XDI at 100-fold dilution also showed a significant increase of MS (3237.24 ± 78.38) by 34.8% compared with blank group (2265.40 ± 109.58) (P < 0.01) (Figure 4(b)). As shown in Figure 4, DHI and MLNI at all concentrations did not have significant changes in any of the parameters studied. The result of HCA assay suggested that the high concentration of FFKSI and XDI may cause mitochondrial damage of HepG2 cells. In addition, for DHI, XDI, MLNI, and FFKSI, the IC 50 of CN was 0.41-fold, 8.6 × 10−3-fold, 30.77-fold, and 2725.78-fold; the IC 50 of MS was 741.85-fold, 3.77-fold, 0.35-fold, and 2725.68-fold; the IC 50 of NA was 404.25-fold, 821.73-fold, 109.64-fold, and 2199.53-fold; the IC 50 of MMP was 5.95 × 10−6-fold, 220.91-fold, 345.43-fold, and 4.95 × 107-fold; the IC 50 of PMP was 660.16-fold, 90.01-fold, 268.64-fold, and 1.31 × 109-fold.


Safety evaluation of chinese medicine injections with a cell imaging-based multiparametric assay revealed a critical involvement of mitochondrial function in hepatotoxicity.

Wang M, Liu CX, Dong RR, He S, Liu TT, Zhao TC, Wang ZL, Shen XY, Zhang BL, Gao XM, Zhu Y - Evid Based Complement Alternat Med (2015)

Representative images of the HCA analysis of HepG2 cells for the evaluation of nuclei number and morphology stained by 4 dyes in control (a, f, k, p, and u), Danhong injection (DHI) 100-fold dilution (b, g, l, q, and v), Xiangdan injection (XDI) 100-fold dilution (c, h, m, r, and w), Mailuoning injection (MLNI) 100-fold dilution (d, i, n, s, anad x), and Fufangkushen injection (FFKSI) 100-fold dilution (e, j, o, t, and y) groups. The cell number (CN) decreased by XDI 100-fold dilution and FFKSI 100-fold dilution (c, e). The intensity of Rhodamine 123 (green) was lower in FFKSI-treated group than control group indicating that the MMP decreased (o). Note: IC 50 of cell parameters associated with cell number (CN) was IC 501 and IC 50 of cell parameters associated with nuclear area (NA) was IC 502.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Representative images of the HCA analysis of HepG2 cells for the evaluation of nuclei number and morphology stained by 4 dyes in control (a, f, k, p, and u), Danhong injection (DHI) 100-fold dilution (b, g, l, q, and v), Xiangdan injection (XDI) 100-fold dilution (c, h, m, r, and w), Mailuoning injection (MLNI) 100-fold dilution (d, i, n, s, anad x), and Fufangkushen injection (FFKSI) 100-fold dilution (e, j, o, t, and y) groups. The cell number (CN) decreased by XDI 100-fold dilution and FFKSI 100-fold dilution (c, e). The intensity of Rhodamine 123 (green) was lower in FFKSI-treated group than control group indicating that the MMP decreased (o). Note: IC 50 of cell parameters associated with cell number (CN) was IC 501 and IC 50 of cell parameters associated with nuclear area (NA) was IC 502.
Mentions: Five fixed concentrations (1000-, 800-, 500-, 300-, 100-fold dilution) of four injections, DHI, XDI, FFKSI, and MLNI were selected. As shown in Figure 3, the representative images were captured by the HCA method established for typical cytotoxic effects on hepatocyte induced by FFKSI 100-fold dilution, XDI 100-fold dilution, MLNI 100-fold dilution, and DHI 100-fold dilution. It is noteworthy that 1000-, 800-, 500-, 300- (P < 0.05), and 100-fold dilution (P < 0.01) of FFKSI significantly decreased the MMP by 11.1%–22.8% as compared to blank group (Figure 4(d)). XDI and FFKSI at 100-fold dilution caused a significant decrease of CN by 50.8% and 48.4% (P < 0.05), respectively (Figure 4(a)). XDI at 100-fold dilution also showed a significant increase of MS (3237.24 ± 78.38) by 34.8% compared with blank group (2265.40 ± 109.58) (P < 0.01) (Figure 4(b)). As shown in Figure 4, DHI and MLNI at all concentrations did not have significant changes in any of the parameters studied. The result of HCA assay suggested that the high concentration of FFKSI and XDI may cause mitochondrial damage of HepG2 cells. In addition, for DHI, XDI, MLNI, and FFKSI, the IC 50 of CN was 0.41-fold, 8.6 × 10−3-fold, 30.77-fold, and 2725.78-fold; the IC 50 of MS was 741.85-fold, 3.77-fold, 0.35-fold, and 2725.68-fold; the IC 50 of NA was 404.25-fold, 821.73-fold, 109.64-fold, and 2199.53-fold; the IC 50 of MMP was 5.95 × 10−6-fold, 220.91-fold, 345.43-fold, and 4.95 × 107-fold; the IC 50 of PMP was 660.16-fold, 90.01-fold, 268.64-fold, and 1.31 × 109-fold.

Bottom Line: The safety of herbal medicine products has been a widespread concern due to their complex chemical nature and lack of proper evaluation methods.Our results suggested that the reported hepatotoxicity by one of the drugs, Fufangkushen injection, could be attributed at least in part to the interference of mitochondrial function in human HepG2 cells by some of its constituents.This method should be useful for both preclinical screen in a drug discovery program and postclinical evaluation of herbal medicine preparations.

View Article: PubMed Central - PubMed

Affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Nankai District, Tianjin 300193, China ; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tianjin 300457, China.

ABSTRACT
The safety of herbal medicine products has been a widespread concern due to their complex chemical nature and lack of proper evaluation methods. We have adapted a sensitive and reproducible multiparametric cell-based high-content analysis assay to evaluate the hepatic-safety of four Chinese medicine injections and validated it with classical animal-based toxicity assays. Our results suggested that the reported hepatotoxicity by one of the drugs, Fufangkushen injection, could be attributed at least in part to the interference of mitochondrial function in human HepG2 cells by some of its constituents. This method should be useful for both preclinical screen in a drug discovery program and postclinical evaluation of herbal medicine preparations.

No MeSH data available.


Related in: MedlinePlus