The Antitumor Activities Of Sapindus Water Extracts

Sapindus, also was known as soapberry, fruit contain a great deal of saponin and sesquiterpene glucosides. The extract of sapindus has strong biological activities such as anti-inflammatory, sterilization and antitumor. Currently, the researches on material basis of antitumor effect on sapindus extract were not systematic, most of which were in vitro experiments. Therefore in this paper the antitumor activities of compositions from sapindus water extract were carried out.The grading separation of sapindus water extract. The concentrated solution of sapindus pericarp water extract was extracted by same volume ethyl acetate for two times, removed the residual ethyl acetate by heating, and then separated by X-5 and AB-8 macroporous resin column. Two elution parts were obtained by water and 95% ethanol(volume fraction) respectively. The water elution part was named as Fraction I. The 95% ethanol elution part was further eluted by water, 30% ethanol and 95% ethanol from MCI gel column and Fraction II, III and IV were obtained respectively.Antitumor activity of sapindus fractions in vitro. The in vitro effect was determined by MTT assay, which indicated that Fraction II and IV had no obvious growth inhibition activity on human breast normal cell lines MDA-kb2. Fraction II and IV also had strong growth inhibition activity on human breast cancer cell lines MCF-7(estrogen receptor positive), MDA-MB-231(estrogen receptor negative), and human breast ductal cancer cell lines MDA-MB-435 s respectively and showed a significant dose-dependent relationship. The IC50 of Fraction II against these three cell lines were 49.90、39.55 and 33.11 μg/mL, which of Fraction IV were 40.35, 32.18 and 30.06 μg/m L respectively.Antitumor activity of sapindus fractions in vivo. The in vivo antitumor effects of Fraction II, III and IV were carried out by establishing mice models of 4T1 mice breast cancer. The results showed that, three fractions could make mice tumor cells necrosis such as karyopyknosis and nuclear fragmentation, and had no obvious effect on the immune system of mice; Fraction II and IV had excellent antitumor effects at a dose of 162.5 mg/kg/d with inhibition rates of 40.43% and 38.77% respectively. Analysis by flow cytometry revealed that three fractions could induce significant apoptosis in tumor cells compared with the negative control(P<0.05) with an apoptosis rates of 56.19%, 1.91% and 56.70% respectively. Further immunohistochemical staining revealed that Fraction II caused the Bax positive cells in the integral optical density(IOD) value significantly higher than that in negative control(P<0.05), and the Bcl-2 positive cells IOD was significantly lower than that in negative control(P<0.05), indicated that Fraction II increased the pro-apoptotic proteins Bax expression and decreased the expression of anti-apoptotic protein Bcl-2 in mice tumor to induce cell apoptosis. Fractions IV only caused the Bcl-2 positive cells IOD value significantly lower than that in negative control(P<0.05), indicating that the Fraction IV may induce cell apoptosis by decreasing the expression of Bcl-2.The further separation and structure identification of sapindus anti-breast cancer active fraction. Using the ODS chromatography column to further separate Fraction II, two monomer compounds were acquired, namely compound 1 and 2, and they were belong to one sugar chain of sesquiterpene glucoside by HPLC-TOF/MS, 1D and 2D NMR analysis. The in vitro effect was determined by MTT assay, which indicated that Component 1 and 2 had medium intensity growth inhibition activities on MCF-7, MDA-MB-231, and MDA-MB-435 s respectively. The IC50 of Component 1 were 128.28, 154.01 and 134.21 μg/m L, while Component 2 were 120.73, 134.75 and 133.54 μg/mL respectively which were not a patch on the IC50 of Fraction II, and indicated that further research needed.