Seed oil of Brucea javanica induces apoptosis through the PI3K/Akt signaling pathway in acute lymphocytic leukemia Jurkat cells / 中国天然药物

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3β was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.

Formulation development and evaluation of gastroretentive floating beads with Brucea javanica oil using ionotropic gelation technology / 中国天然药物

In the present study, a gastric retention floating system for Brucea javanica oil, composed of alginate and carrageenan, was prepared using ionotropic gelation. Parameters for floatability, drug load, encapsulation efficiency, bead morphology, in vitro release, and in vivo gastric retention were evaluated. The optimized formulation via Box-Behnken design consisted of 1.7% alginate (W/V), 1.02% carrageenan (W/V), 1.4% CaCO (W/V), and a gelling bath of pH 0.8. The alginate-carrageenan-Brucea javanica oil beads had a porous structure and exhibited up to 24 h of in vitro floatability with a load capacity of 45%-55% and an encapsulation efficiency of 70%-80%. A 6-h sustained release was observed in vitro. The beads had a prolonged gastric retention (> 60% at 6 h) in fasted rats, compared to non-floating beads (15% at 6 h), as measured by gamma scintigraphy with single-photon emission tomography/computed tomography (SPET/CT). In conclusion, the alginate-carrageenan-Brucea javanica oil system showed enhanced oil encapsulation efficiency, excellent floating and gastric retention abilities, and a favorable release behavior.

Effect of Brucea javanica oil emulsion on proliferation, migration and autophagy of non-small cell lung cancer A549 cells and the underlying mechanisms / 中南大学学报(医学版)

To investigate the effect of Brucea javanica oil emulsion on proliferation, migration and autophagy of non-small cell lung cancer A549 cells.
 Methods: First, A549 cells were divided into a control group and a low, medium or high dose of Brucea javanica oil emulsion groups (0, 2.5, 5.0 or 10.0 mg/mL); then, the cells were divided into a 3-MA+Brucea javanica oil emulsion group and a Brucea javanica oil emulsion group in the presence or absence of 3-methyl adenine (3-MA). Cell counting kit-8 (CCK-8) and clone formation assay were used to detect cell proliferation, while the wound scratch and Transwell assay were used to measure cell migration. Cell immunofluorescence and Western blot were used to analyze autophagy.
 Results: Compared with the control group, the numbers of cell proliferation and colony-formation, the relative cell migration rate and numbers of trans-membrane cells were reduced in a dose-dependent manner in the Brucea javanica oil emulsion groups (all P<0.05). Meanwhile, compared with the control group, the aggregation of microtubule associated protein 1 light chain3 (LC3) green fluorescence and the LC3-II/LC3-I ratios were increased, while p62 level was decreased (all P<0.05) in the high dose group. Compared with the Brucea javanica oil emulsion group (5.0 mg/mL), the cell proliferation, numbers of cell clone formation, cell migration rate and numbers of Transwell transmembrane cells were increased in the 3-MA+Brucea javanica oil emulsion group (all P<0.05).
 Conclusion: Brucea javanica oil emulsion can promote the autophagy of non-small cell lung cancer A549 cells and inhibit the cell proliferation and migration.

Formulation development and evaluation of gastroretentive floating beads with Brucea javanica oil using ionotropic gelation technology / 中国天然药物

In the present study, a gastric retention floating system for Brucea javanica oil, composed of alginate and carrageenan, was prepared using ionotropic gelation. Parameters for floatability, drug load, encapsulation efficiency, bead morphology, in vitro release, and in vivo gastric retention were evaluated. The optimized formulation via Box-Behnken design consisted of 1.7% alginate (W/V), 1.02% carrageenan (W/V), 1.4% CaCO (W/V), and a gelling bath of pH 0.8. The alginate-carrageenan-Brucea javanica oil beads had a porous structure and exhibited up to 24 h of in vitro floatability with a load capacity of 45%-55% and an encapsulation efficiency of 70%-80%. A 6-h sustained release was observed in vitro. The beads had a prolonged gastric retention (> 60% at 6 h) in fasted rats, compared to non-floating beads (15% at 6 h), as measured by gamma scintigraphy with single-photon emission tomography/computed tomography (SPET/CT). In conclusion, the alginate-carrageenan-Brucea javanica oil system showed enhanced oil encapsulation efficiency, excellent floating and gastric retention abilities, and a favorable release behavior.

Research of Brucea javanica against cancer / 中国结合医学杂志

Brucea javanica, a Chinese herbal medicine, combined with conventional anticancer modalities, has been widely used for treatment of various cancers. Based on researches over the last decades, authors briefly summarized its active constituents, molecular mechanisms and clinical application for cancer treatment.

Effect of Brucea javanica fruit oil emulsion combined cisplatin on the growth inhibition of transplanted tumor in human ovarian cancer SKOV3 nude mice: an experimental study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the growth inhibition effect of Brucea javanica Fruit Oil Emulsion (BJFOE) on human ovarian caner SKOV3 cells and the transplanted tumor of SKOV3 nude mice.</p><p><b>METHODS</b>Growth inhibition effects of different concentrations BJFOE alone or its combination with cisplatin on human ovarian cancer cell SKOV3 were measured using MTT method. The orthotopic transplantation tumor model of human ovarian cancer SKOV3 cell lines was established in nude mice. Totally 32 ovarian cancer nude mice were randomly divided into 4 groups, i.e., the blank control group (Group A), the BJFOE group (Group B), the BJFOE combined Cisplatin group (Group C), and the Cisplatin control group (Group D), 8 in each group. Mice in Group A were intraperitoneally injected with normal saline (0.2 mL/ 20 g), once per two days. Mice in Group B were intraperitoneally injected with BJFOE (0.2 mL/20 g), once per two days. Mice in Group C were intraperitoneally injected with cisplatin (3 mg/kg) 0.2 mL on the first day, and intraperitoneally injected with BJFOE on the second day. Mice in Group D were intraperitoneally injected with cisplatin (3 mg/kg) 0.2 mL, once per two days. All mice were injected for six times, and sacrificed 48 h after the last injection. The lesion formation of the abdominal tumor tissue was observed. Tumor specimens were obtained to perform HE staining. Expression levels of MRP-1/CD9 and integrinα-5 were detected using Western blot.</p><p><b>RESULTS</b>The inhibition of BJFOE was time-dose depend- ently correlated with its inhibition effect of SKOV3 cells. The inhibition effect of BJFOE in combination of cisplatin was significantly superior to that of using any of the two drugs alone. Western blot results showed expression levels of MRP-1/CD9 and integrinα-5 were up-regulated in Group B and Group D with statistical difference (P < 0.05). But they were down-regulated in Group C with statistical difference (P < 0.05).</p><p><b>CONCLUSIONS</b>Intraperitoneal injecting BJFOE was feasible and effective for treating ovarian cancer. BJFOE also could inhibit the invasion and migration of tumor cells targeting at MRP-1/CD9 and integrinα-5. But its specific anti-tumor mechanism was not clearly probed.</p>

A new indole alkaloid from the stems of Brucea mollis / 药学学报

Eight compounds were isolated from the stems of Brucea mollis by various chromatographic techniques such as column chromatography on silica gel and Sephadex LH-20, and preparative HPLC, and their structures were elucidated as bruceolline O (1), 1-(1-beta-glucopyranosyl)-1H-indole-3-carbaldehyde (2), canthin-6-one (3), 11-hydroxycanthin-6-one (4), 9-methoxycanthin-6-one (5), 4-methoxycanthin-6-one (6), infractin (7), and beta-carboline-1-propionic acid (8). The cytotoxic activities of compounds 1-8 against HCT-8 and A549 human cell lines were determined, but none of them exhibited significant activity (IC 50 > 10 micromol x L(-1)). Among them, compound 1 is a new indole alkaloid, and compounds 2 and 5-7 were isolated from this plant for the first time.

Chemical constituents from stems of Brucea mollis and their cytotoxic activity / 中国中药杂志

Ten compounds were isolated from the stems of Brucea mollis by various chromatographic techniques such as column chromatography on silica gel and Sephadex LH-20, and preparative HPLC, and their structures were elucidated as deacetylated isobrucein B (1), indaquassin X (2), cleomiscosin A (3), cleomiscosin B (4), (+)-lyoniresinol (5), (+)-epipinoresinol(6), (+)-pinoresinol (7), (+)-syringaresinol (8), 4,5-dihydroblumenol A (9) and adenosine (10) on the basis of spectroscopic data analysiS. All compounds were obtained from this plant for the first time, moreover, compound 1 was a new natural product. Compound 2 showed significant cytotoxic activities against the human cell lines HT-29, HepG2, BGC-823 and SKOV3 with IC50 values of 0.84-3.97 micromol x L(-1).

Effects of Brucea javanica oil emulsion on human papilloma virus type 16 infected cells and mechanisms research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the in vitro inhibitive effect and underlying mechanisms of Brucea Javanica oil emulsion (BJOE) on human papilloma virus (HPV) type 16 infected cells.</p><p><b>METHODS</b>The HPV16 E61E7 immortalized human ectocervical Ect1/E6E7 cell line and the CaSki cell line were selected as the in vitro models of premalignant cervical lesion and cervical cancer respectively. After treated with BJOE at different concentrations (5, 10, 20, and 40 microg/mL) at the operation time points (24, 48, and 72 h), the effects of BJOE on proliferative activities were measured by MTT assay. The morphologic changes of cell apoptosis stained with Hochest 33,258 were observed by fluorescence microscope. The effect on the cell apoptosis rate was analyzed by Annexin V-FITC/PI double-labeled flow cytometry. The mRNA expressions of HPV16 E6 and E7 were determined by semi-quantitative RT-PCR. The protein expressions of HPV16 E6, E7 oncogene, and specifically interacted p53, Rb antioncogene were stained by immunocytochemical staining (Elivison two-step procedure).</p><p><b>RESULTS</b>(1) The proliferative activities of the Ect1/E6E7 cell and the CaSki cell treated with BJOE at different concentrations (5, 10, 20, and 40 p g/mL) at the operation time points (24, 48, and 72 h) were obviously inhibited, showing dose- and time-dependent manners (P <0.05). (2) Typical changes of apoptosis were observed in both HPV 16 positive cell lines after treated with BJOE. The cell apoptosis rates increased markedly after being cultured with BJOE at different concentrations (5, 10, and 20 microg/mL) for 48 h (P < 0.05). (3) After treated with BJOE at different concentrations (5, 10, and 20 microg/mL) for 48 h, the HPV16 E6 and E7 mRNA relative expressions in both HPV 16 positive cell lines decreased significantly (P < 0.05). (4) After treated with BJOE at different concentrations (5, 10, and 20 microg/ mL), the expressions of HPV16 E6, E7, and mutant p53 protein decreased gradually (P < 0.05), while the Rb protein expression increased gradually (P < 0.05).</p><p><b>CONCLUSIONS</b>BJOE showed obvious in vitro inhibitory effects on HPV type 16 infected cells. Its underlying mechanisms might be possibly associated with down-regulating expressions of HPV16 E6 and E7 oncogenes.</p>

Meta-analysis on treatment of non-small cell lung cancer with brucea javanica oil emulsion in combination with platinum-contained first-line chemotherapy / 中国中药杂志

<p><b>OBJECTIVE</b>To study the efficacy and safety of brucea javanica oil emulsion (BJOE) combining platinum-contained first-line chemotherapy in treating non small cell lung cancer (NSCLC).</p><p><b>METHOD</b>Cochrane library, PubMed, EMBASE, VIP, CBM and CNKI were searched through computers. The search was finished in February, 2011. Randomized controlled trials (RCTs) of BJOE combining platinum-contained first-line chemotherapy were included. Two researchers extracted data and assess literature quality separately,and made a meta-analyses by RevMan 5.1.2 software.</p><p><b>RESULT</b>Totally 22 RCTs involving 1512 patients were included. The Meta-analysis showed that compared with the pure application of platinum-contained first-line chemotherapy,the combination of BJOE and chemotherapy can enhance the near-term curative effect (RR = 1. 31, 95% CI: 1.18-1.45, P < 0. 000 01), improve the quality of life (RR = 1.78, 95% CI: 1.51-2. 09, P < 0.00001) and reduce the suppression of bone marrow (OR = 0.37, 95% CI: 0. 27-0. 51, P < 0.00001) and the gastrointestinal reactions (OR = 0.59, 95% CI: 0.44-0.80, P = 0.0007) ,with an improvement in organism immunity.</p><p><b>CONCLUSION</b>The current evidence indicates that BJOE can enhance the chemotherapeutic effect on NSCLC patients, improve the quality of life and reduce adverse effect of platinum-contained chemotherapeutics and thus it is worth referring in clinic.</p>

Quantitative determination of contents of three components in Brucea javanica by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for quantitative determination of three quassinoids in Brucea javanic.</p><p><b>METHOD</b>The determination was carried out on a phenomenex C18 column (4.6 mm x 250 mm, 5 microm) with gradient elution program of methol-water at a flow rate of 1.0 mL x min(-1), and the detection wavelength was 270 nm.</p><p><b>RESULT</b>Linearites of bruceine D, brusatol and bruceine H were good (r = 0.9996, 0.9996, and 0.9998) in ranges of 2.52-12.60, 2.19-10.95, and 2.91-14.55 microg, respectively. The average recoveries of bruceine D, brusatol and bruceine H were 100.01%, 100.95% and 100.43% respectively, and RSD of the above three compounds were 0.31% (n = 6), 1.7% ( n = 6) and 1.7% (n = 6), respectively.</p><p><b>CONCLUSION</b>The determination results of three batch samples showed that the method was simple, accurate and could be used in the quantitative determination of three components in the B. javanica.</p>

Treatment of middle/late stage primary hepatic carcinoma by Chinese medicine comprehensive therapy: A prospective randomized controlled study / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the efficacy of Chinese medicine comprehensive therapeutic project in treating the middle/late stage primary hepatic carcinoma (PHC).</p><p><b>METHODS</b>With prospective randomized controlled design, 97 patients with PHC were assigned to the test group (49 cases) treated with Chinese medicine comprehensive therapy using Oleum fructus bruceas intervention combining oral intake of Ganji Decoction and external application of Ailitong, and the control group (48 cases) treated with chemotherapeutic agents combining iodized oil chemo-embolization and analgesics. The immediate and long-term efficacy, adverse reaction, pain-relieving initial time (PRIT) and pain-relieving sustained time (PRST) of the treatment, as well as the change in relieving patients' quality of life (QOL) were observed.</p><p><b>RESULTS</b>The difference between the two groups in illness control rate was statistically insignificant (P>0.05), but the adverse reaction occurrence rate in the test group was lesser than that in the control group (P<0.05). PRIT was insignificantly different in the two groups (P>0.05), but the PRST was significantly superior in the test group than that in the control group (10.37+/-2.18 h vs 7.78+/-1.95 h, P<0.01). After treatment, the increased Karnofsky scores in the test group indicated that the patients' somatic activity, symptoms and QOL were improved significantly, which were significantly superior to those in the control group (P<0.05). The survival rate in the two groups was similar at the 3rd month after treatment, but the test group did show superiority in terms of half- and 1-year survival rate (65.9% vs 42.5% and 38.6% vs 18.1%, respectively, P<0.05). The median survival time in the test group was 8.9 months and that in the control group was 5.3 months.</p><p><b>CONCLUSION</b>Chinese medicine comprehensive therapy is an effective treatment for the middle/late stage patients of PHC, and it could extend the PRST, improve the patients' QOL and long-term survival with less adverse reaction.</p>

Effect of oleum fructus brucease injection via bronchial arterial infusion in treating advanced lung cancer / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the clinical effects of bronchial arterial infusion (BAI) with oleum fructus bruceae (OFB) Injection and chemotherapeutic agents (CTA) in treating advanced non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>One hundred and three patients with advanced NSCLC were randomized into 2 groups, the 98 patients in the treatment group treated by BAI with OFB + CTA and the 50 in the control group by BAI with CTA alone. The incidence of adverse reaction, change of tumor size and patients' quality of life (QOL) in the two groups were observed and compared.</p><p><b>RESULTS</b>The objective effective rate (CR + PR) was 63.3% in the treatment group and 46.0% in the control group (P < 0.01); the median survival duration in them was 363 days and 305 days; the 1-year cumulative survival rate was 70.4% and 44.0%, and the QOF improving rate was 83.7% and 62.0% respectively, the difference between groups were all statistically significant (P < 0.01). In addition, the incidence of adverse reactions of digestive symptoms, bone marrow suppression and the hepato-, renal and cardiac toxicities were lower in the treatment group than those in the control group (P < 0.01).</p><p><b>CONCLUSION</b>BAI with OFB + CTA in treating NSCLC could enhance the objective therapeutic effect of simple chemotherapy, as well as raise the QOL and protect immune and medulla function in patients.</p>

Effect of ganji recipe combined with Fructus Bruceae oil emulsion intervention on quality of life in patients with advanced primary hepatic cancer / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe and compare the quality of life (QOL) and survival time in patients with advanced primary hepatic cancer (PHC) after they have been treated by the combination of ganji recipe and interventional therapy with Fructus Bruceae Oil Emulsion (FBE) or by the trans-hepatic arterial chemical embolization (TACE) adopting Seldinger's technique.</p><p><b>METHODS</b>Seventy-seven patients with advanced PHC were randomly assigned to two groups, 37 patients in the control group treated with TACE alone, and 40 in the treatment group with the combined therapy. One therapeutic course was 4 weeks, and the intervention therapy was repeated generally after an interval of 4-6 weeks according to patients'condition ,the average intervention frequency was 2.3 for one case. The QOL and the survival time in patients were evaluated after treatment.</p><p><b>RESULTS</b>Significant difference was shown in the changes of the Karnofsky score between the two groups (P < 0.05). The scores suggested that the improvements in the treatment group, including patients'physical energy enhancing, symptoms alleviating and overall QOL improving, were superior to those in the control group (P < 0.05). No significant difference of the 3-month survival rate between the two groups was found (P > 0.05), but the 0.5- and 1-year survival rate were significantly superior in the treatment group to those in the control group (67.6% vs. 42.4% and 38.2% vs. 16.1%, respectively). The median survival time in the two groups was 8.9 and 5.3 months respectively.</p><p><b>CONCLUSION</b>Combination of ganji recipe and interventional therapy with FBE in treating PHC could improve patients' QOL, raise the 0.5- and 1-year survival rate, and prolong the survival time, suggesting TCM treatment has an affirmative significance for enhancing the therapeutic efficacy and improving the prognosis in patients.</p>

Effects of brucea javanica oil on expression of vascular endothelial growth factor in A549 cell line / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of brucea javanica oil on the expression of vascular endothelial growth factor (VEGF) in A549 cell line.</p><p><b>METHOD</b>A549 cells were incubated with different concentrations of brucea javanica oil (0.5, 1.25, 2.5, 5 g x L(-1) for 48 h respectively. VEGF level in supernatant was determined by VEGF ELISA kits and mRNA expression of VEGF was evaluated by RT-PCR. PMN in health volunteers was treated as control groups.</p><p><b>RESULT</b>Supernatant VEGF protein and mRNA expression were significantly elevated in A549 cells compared with the mononuclear cells (120.73 vs 21.21, P < 0.05). Brucea javanica oil (2.5 g x L(-1)) could reduced supernatant VEGF protein in A549 cells (20.30 vs 120.73, P < 0.05), but had no effect on the expression of VEGF mRNA (1.0230 vs 0.9573). It was found that brucea javanica oil (5 g x L(-1)) significantly reduced VEGF mRNA expression (0.4682 vs 0.9573, P < 0.05).</p><p><b>CONCLUSION</b>Brucea javanica oil can depress the VEGF mRNA expression and secretion in A549 cells, which may be one of the mechanisms of its antitumor effect.</p>

Effect of Fructus Bruceae oil emulsion on cellular immune function and quality of life in patients with non-small cell lung cancer / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Fructus Bruceae oil emulsion (FBE) on cellular immune function (CIF) and quality of life (QOF) in patients with non-small cell lung cancer (NSCLC) after chemotherapy.</p><p><b>METHODS</b>One hundred and fifteen patients with mid-late stage NSCLC were randomly assigned to two groups, the 57 patients in the control group were only treated with chemotherapy of GP regimen, 58 in the treatment group with the chemotherapy of the same regimen and combined with FBE. The clinical efficacy was evaluated after two cycles of chemotherapy.</p><p><b>RESULTS</b>The effective rate was 51.8% and 47.4% in the treatment group and the control group respectively, the difference between them was insignificant (P > 0.05). CIF and QOF in the treatment group were better than those in the control group after chemotherapy respectively (P < 0.01), in the latter, CIF and QOF were desreased after chemotherapy (P < 0.05).</p><p><b>CONCLUSION</b>FBE combined with chemotherapy can improve the cellular immune function and quality of life in patients with mid-late stage NSCLC.</p>

Experimental studies on the apoptosis of HL-60 cells induced by Brucea javanica oil emulsion / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the inducing effect of Brucea javanica on the apoptpsis of HL-60 cells.</p><p><b>METHOD</b>HL-60 cells were treated with Brucea javanica 1:100, 1:40, 1:20 (v/v) respectively for 6 h and DNA agarose gel electrophoresis, flow cytometry, fluorescence microscope, electron microscope were used to observe the apoptosis inducing effect of Brucea javanica.</p><p><b>RESULT</b>DNA ladder was seen in the 1:40 group. The apoptosis cell percentages of 1:40 and 1:20 group were 86.8% and 97% respectively. Cells of 1:40 group showed obvious apoptosis character under fluorescence microscope. Cells were induced apoptosis in 1:20 and 1:40 Brucea javanica under electron microscope.</p><p><b>CONCLUSION</b>1:20 and 1:40 Brucea Javanica showed obvious apoptosis inducing effect of HL-60.</p>