Chinese Herbal Medicine, Guilu Erxian Glue, as Alternative Medicine for Adverse Side Effects of Chemotherapy in Doxorubicin-Treated Cell and Mouse Models

Figures

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  Figure 1. Chromatographic fingerprint analysis for the herbal formula Guilu Erxian Glue (GEG). GEG is a combination of traditional Chinese medicine with four main ingredients of ginseng (Radix Ginseng), wolfberry (Fructus Lycii), tortoise plastron (Carapax et Plastrum Testudinis), and antler (Cornu Cervi) from a famous Chinese prescription. (a) 3D-HPLC fingerprint of Ginsenoside (200–400 nm, 0–65 min). (b) 3D-HPLC fingerprint of Betaine and Nucleosides (200–400 nm, 0–30 min). Chromatographic fingerprint analysis was conducted through the HPLC and LC/MS. Abbreviations: GEG, Guilu Erxian Gum; AU, arbitrary perfusion units; and HPLC, high-performance liquid chromatography.

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  Figure 2. Free-radical scavenging activity and HIG-82 cell viability after GEG treatment. (a) Quantified DPPH radical scavenging activity at various concentrations of GEG (N = 3). (b) Quantified relative HIG-82 cell viability at various concentrations of GEG with (+DOX) and without (−DOX) DOX treatment (N = 3 for each). Values are expressed as the mean ± SEM (∗∗p < 0.01, ∗p < 0.05, two-way ANOVA followed by a Student–Newman–Keuls multiple-comparison posttest). Abbreviations: GEG, Guilu Erxian Gum; DPPH, 1,1-diphenyl-2-picrylhydrazyl; DOX: doxorubicin; SEM, standard error of the mean; and ANOVA, analysis of variance.

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  Figure 3. Effects of GEG treatment on the body weight and exhaustive swimming time of mice with and without DOX treatment. (a) Comparison of quantified body weight among the mice treated with sham, GEG, DOX, and DOX + GEG (N = 8 for each group). (b) Comparison of EST among mice treated with sham, GEG, DOX, and DOX + GEG (N = 8 for each group). Values are expressed as the mean ± SEM (∗∗p < 0.01, ∗p < 0.05, two-way ANOVA followed by a Student–Newman–Keuls multiple comparison posttest). Abbreviations: GEG, Guilu Erxian Gum; EST, exhaustive swimming time; DOX, doxorubicin; SEM, standard error of the mean; ANOVA, analysis of variance.

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  Figure 4. Effects of GEG treatment on subcutaneous microcirculation and RBC count in mice treated with and without DOX. (a) Dorsal imaging of subcutaneous microcirculation performed using moorFLPI laser Doppler imaging (left) and comparison of quantified subcutaneous blood flow (right) among mice treated with sham, GEG, DOX, and DOX + GEG (N = 8 for each group). (b) Peripheral blood smear analysis (left) and comparison of the quantified number of RBCs (right) among mice treated with sham, GEG, DOX, and DOX + GEG (N = 8 for each group). WBCs are marked with arrows. Values are expressed as the mean ± SEM (∗∗p < 0.01, ∗p < 0.05, one-way ANOVA followed by a Student–Newman–Keuls multiple-comparison posttest). Abbreviations: AU, arbitrary unit; RBC: red blood cell; WBC: white blood cell; GEG, Guilu Erxian Gum; DOX, doxorubicin; SEM, standard error of the mean; and ANOVA, analysis of variance.

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  Figure 5. Effects of GEG treatment on antioxidant-stress-related SOD2 expression in the myocardial tissue of mice treated with and without DOX. Horizontal and longitudinal cross sections of IHC staining results showing myocardial SOD2 expression in mice treated with sham, GEG, DOX, and DOX + GEG. Bar scale = 30 μm. Abbreviations: HS, horizontal cross sections; LS, longitudinal cross sections; SOD2, superoxide dismutase 2; GEG, Guilu Erxian Gum; and DOX, doxorubicin.

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  Figure 6. Effects of GEG treatment on inflammation-related TNF-α expression in the myocardial tissue of mice with and without DOX treatment. Horizontal and longitudinal cross sections of IHC staining showing myocardial TNF-α expression in mice treated with sham, GEG, DOX, and DOX + GEG. Bar scale = 30 μm. Abbreviations: HS, horizontal cross sections; LS, longitudinal cross sections; TNF-α, tumor necrosis factor alpha; GEG, Guilu Erxian Gum; and DOX, doxorubicin.

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  Figure 7. Effects of GEG treatment on bone density in the tibia tissue of mice with and without DOX treatment. (a) Cross-sectional imaging of the tibia and trabecular bone among mice treated with sham, GEG, DOX, and DOX + GEG performed by high-resolution microcomputed tomography. Bar scale = 100 μm. (b) Comparison of the quantified bone volume percentage of the tibia among mice treated with sham, GEG, DOX, and DOX + GEG (N = 3 for each group). Values are expressed as the mean ± SEM (∗∗p < 0.01, ∗p < 0.05, one-way ANOVA followed by a Student–Newman–Keuls multiple-comparison posttest). Abbreviations: GEG, Guilu Erxian Gum; DOX, doxorubicin; SEM, standard error of the mean; and ANOVA, analysis of variance.