Efficacy of Jasminum subtriplinerve Extract against 7,12-Dimethylbenz[ a]anthracene-Induced Cancer in Mice

Abstract

Jasminum subtriplinerve Blume tea is a traditional Vietnamese medicine used to treat impetigo, menstruation issues, and painful menstrual hematometra. Previous studies have shown that extracts and isolated compounds from J. subtriplinerve possess diverse pharmacological properties, such as antioxidant, antibacterial, and antidiabetic effects. However, their potential anticancer effects and underlying mechanisms of action have not been clear. Here, we examined the effects of J. subtriplinerve extracts against three human cancer cell lines. We also conducted in vivo analyses using a mouse model of 7,12-dimethylbenz[a]anthracene-induced breast cancer, including an investigation of changes in histological sections. The effect of the J. subtriplinerve ethyl acetate fraction on cytokine levels (IL-2, PGE2, TNF-α) in serum was determined using ELISA kits. Results showed that the ethyl acetate (EtOAc) fraction had the highest anti-proliferative activity (IC50 = 13.7 mg/ml) against the breast cancer (MCF-7) cell line, while the butanol (BuOH) and water fractions did not show any anticancer effects. Additionally, the EtOAc fraction at a dose of 14.4 mg/kg was able to elevate IL-2 levels and suppress the expression of PGE2 in the serum of mice. A remarkable decrease in the percentage of death and tumor incidence in mice was achieved following treatment with the EtOAc fraction at a dose of 14.4mg/kg. No abnormal parameters in blood were observed in the J. subtriplinerve treatment groups. These results suggest that J. subtriplinerve, when used as tea or a functional food, is nontoxic and has clear chemopreventive effects against breast cancer.

Keywords: DMBA; IL-2; Jasminium subtriplinerve Blume; breast cancer.

Fig. 1. Screening secondary metabolites from an MeOH extract of J. subtriplinerve using LC-QTOF MS/MS in positive mode.

(A) Positive mass (MS) chromatogram. (B) Ultraviolet (UV; 254 nm) chromatogram. (C) Ultraviolet (UV; 210 nm) chromatogram of the MeOH extract from J. subtriplinerve.

Fig. 2. Anti- cancer effects of J. Subtriplinerve extract (EtOH) and EtOH fractions in-vitro experiment.

EtOH extract and its fractions was treated on three human cancer cell lines in 48 h. The cytotoxicity of J. Subtriplinerve on cancer cell lines was assessed using the MTT assay. IC₅₀ values was calculated (A). Population doubling times are shown (B). Data are presented as the means ± SD of three independent experiments.

Fig. 3. Effects of J. subtriplinerve fractionated extract (EtOAc) on clinical symptoms in a mouse model of DMBA-induced cancer.

Clinical symptoms were observed during the experimental period. (A) Experimental scheme. (B) Survival rate. (C) Number of mice suspected to have tumors and confirmed via H&E analysis. (D) Cancer incidence. Data are presented as means ± standard deviations of 15 mice per group. Statistical significance was assessed via one-way ANOVA followed by Bonferroni’s post hoc test; *p < 0.05, **p < 0.01, vs. group 2.

Fig. 4. Changes in histological sections.

Histological sections of mouse mammary tissues and ovarian tissues were viewed under 200 × magnification after H&E staining. (A) Groups 1 and 4 showed normal mammary gland structures, with small mammary ducts surrounded by a small amount of fibrous connective tissue. Group 2 showed various histopathological changes such as hyperplasia in some ducts, mild ductal proliferations, and focal epithelial hyperplasia with enlarged, hyperchromatic nuclei. Group 5 exhibited dysplastic mammary glands, an increase in channel number, and irregular cell division. (B) Normal ovarian structure in groups 1 (controls) and 4. Group 2 displayed diffuse tumors that disrupted the normal ovarian histological structure. Cells were large and had irregular nuclei, coarse chromatin, and a high nucleus-tocytoplasm ratio. They also showed a high rate of division.

Fig. 5. Effects of extracts on cytokines and lymphocytes.

Treated mice were dissected 1 day after the last treatment. Blood and draining lymph nodes were collected for tests. (A) The gating of CD3, CD8, CD4 T cells by flow cytometer analysis, (B) percentage CD3 T cells, (C) percentage CD8 T cells and (D) percentage CD4 T cells were counted by CD3-CD4-CD8 test. The level of (E) IL-2 and (F) PGE2, (G) IL-6 expression in serum were measured by ELISA. Data are presented as means ± standard deviations of three mice per group. Statistical significance was assessed via one-way ANOVA followed by Bonferroni’s post hoc test; *p < 0.05, **p < 0.01, vs. group 2.

Fig. 6. Dose toxicity effects.

Female mice were administered J. subtriplinerve daily for 4 weeks and clinical observations were made. (A) Changes in hematological results. (B) Levels of liver marker enzymes. (C) Changes in body weight.

Fig. 7. Molecular docking simulation of acteoside with 3ERT.

(A) Three-dimensional binding interactions and (B) two-dimensional diagram of observed ligand–receptor interactions between acteoside and breast cancer protein (3ERT).