| Seed oil of Brucea Javanica is extracted from the seeds of herb medicine Brucea Javnica L. Merr by petroleum ether, and its emulsion is emulsified by soybean phospholipids (BJOE). Brucea Javanica oil emulsion (BJOE) was approved in 1978, and it has been used clinically to treat carcinomas for many years in China, including lung cancer, prostate cancaer and gastrointestinal cancer. However, its potential as a treatment for acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) has not been evaluated. In this thesis, we first examined the components of BJOE and elucidated its anti-cancer components. Then, the anti-cancer effect of BJOE was investigated in vitro and in vivo, and its underlying mechanisms were also investigated.Our results showed that, BJOE contains six components, including linoleic acid (44.85%), oleic acid (29.24%), hexadecanoic acid (11.75%), octadecanoic acid (5.45%), linolenic acid (4.22%) and phenol (0.59%). Among them, oleic acid and linoleic acid are the active anti-leukemia components. According to the proportion of BJOE, oleic acid (37.5,75 and 150μg/ml) and linoleic acid (56.25,112.5,225μg/ml) can decrease the cell viabilities. The mixture of oleic acid and linoleic acid (low:37.5μg/ml+56.25μg/ml; midde:75μg/ml +112.5μg/ml; high:150μg/ml+225μg/ml) also showed cytotoxic effect on leukemia cells, and the effect is almost like the BJOE. After the mixture of oleic acid and linoleic acid treated, the U937 and HL-60 cells showed apoptotic characteristics. These results suggest that, oleic acid and linoleic acid are the active components of BJOE.BJOE (67.5,125,250 and 500μg/ml) had significant cytotoxic effect on U937, NB4, HL-60 and Jurkat cells and the IC50 concentrations are 265.4,217.5,312.7和329.9μg/ml, respectively. The sensitivity of these cells to BJOE are NB4>U937>HL-60.Then we further investigated the underlying mechanisms. The increase of subGl phase, cleaved PARP and DNA ladder showed that BJOE can induce apoptosis of U937 and HL-60 cells. The ROS release, mitochondrial membrane potential collapse, down-regulation of c-FLIP, involvement of Bcl-2 family proteins and activation of caspases were all involved in the apoptosis process. Pretreatment with NAC or CAT can reverse the ROS release and MMP collapse induced by BJOE, but the apoptosis induced by BJOE can not be reversed. These results suggest that, the mitochondrial pathway and death receptor pathway were all involved in the apoptosis induced by BJOE.For the AML patients'leukemia cells, BJOE can increase the subG1 phase cells and cause PARP cleavage. This suggests that BJOE has strong apoptotic effect on AML patient's leukemia cells.Our further study found that, the apoptotic effects of BJOE on peripheral blood lymphocytes were much smaller than AML cell lines. Under the same circumstances, the cell numbers of PBLs cells in subGl phase and the cell numbers with low MMP after BJOE treated were much lowere than that in AML cells. These results show that, the cytotoxic effect of BJOE on PBLs is much smaller than the AML cell lines.Intravenously injected BJOE (5.5,16.5和49.5mg/kg)for seven days can inhibitor tumor growth in p388 mouse model and BJOE did not affect the thymus index of mouse significantly. However, BJOE can increase the spleen index. BJOE can inhibit U937 tumor growth in the xenograft mouse model. These results suggest that BJOE has strong anti-leukemia effect in vivo and it may increase the immunological function of the mouse.Our results found that BJOE has strong anti-leukemia effect in vitro and in vivo. Its underlying mechanism was to induce apoptosis in leukemia cell lines. BJOE also has strong apoptotic effect on the AML patient's leukemia cells. The apoptotic effect of BJOE on PBLs was much smaller than that on the AML cell lines. BJOE shows therapeutic role in the treatment of AML, and this study shows evidence for BJOE in the clinical treatment of AML.
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