Anti-leukaemic and chemotherapy supportive effects of berberis vulgaris l. Fruit crude extract on in vitro and in vivo models
Cancer is one of the most deadly diseases affecting human, and the second cause of death accounting for about 12.8% worldwide. Among the prominently worldwide cancer-affecting people is leukaemia. The disease is characterized by increasing of immature white blood cells called "blasts",...
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| Format: | Thesis |
| Language: | English |
| Published: |
2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/64035/1/IB%202015%2015IR.pdf http://psasir.upm.edu.my/id/eprint/64035/ |
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| Summary: | Cancer is one of the most deadly diseases affecting human, and the second cause of
death accounting for about 12.8% worldwide. Among the prominently worldwide
cancer-affecting people is leukaemia. The disease is characterized by increasing of
immature white blood cells called "blasts", and is estimated to account for about 3% of
incident cases for all kind of cancer worldwide. There are different kinds of treatment
methods for leukaemia including chemotherapy, which is known to have side effects.
However, herbal therapy is considered to have fewer side effects. Hence, the used of
Berberis vulgaris crude extract (BVFCE) in treating leukaemia is highlighted. Barberry
is an herb derived from Berberidaceae shrub and is known as treat cancer. Total
flavonoid and phenolic content (TFC and TPC) and DPPH assay were analyzed by
spectrophotometer to determine the antioxidant activity of the Barberry. BVFCE was
evaluated for its anti-leukaemic activities against two leukaemic cell lines; Human
Promyelocytic (HL-60) and Mouse Myelomonocytic (WEHI-3) versus the normal
mouse fibroblast cell (3T3) in both in vitro and in vivo leukaemic model. For the in
vitro method, cytotoxic effects of BVFCE were evaluated using MTT assay to
determine the IC50 values and compare with non-leukaemic cell line (The 3T3 cell).
Observation of the morphological changes in the treated leukaemic cell lines with
BVFCE and Doxorubicin (Doxo) was examined using inverted microscope. Moreover,
real time PCR was used to amplify mRNA and determine the expression of some
specific related genes in leukaemia. Whereas, for the in vivo model, leukaemia was
induced in male BALB/c mice using mouse leukaemic cell line (WEHI-3) and
treatment was carried out using different concentrations of BVFCE and chemotherapy
drug, separately for comparison. After the treatment period (5 weeks), the mice were
fasted overnight and sacrificed for biochemical, haematological and histological
analyses of their liver, kidney, heart and spleen. All data were analyzed using one-way
ANOVA followed by Duncan's multiple post hoc tests. Differences between groups
were considered significantly when the p value is less than 0.05. Results showed that
BV100 has a higher level of total phenolic and flavonoids contents as well as
antioxidant activity as compared with other concentrations (p<0.05). Biochemical assay
on alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine
aminotransferase (ALT) showed significantly lower level of these markers in control
and treated groups as compared to cancer group (p<0.05). Leukaemic group showed significantly higher total serum than other groups as well (p<0.05), which indicated the
possible beneficial effect of BVFCE treatment on the above named markers.
From the haematological finding, the total white blood cell (WBC) count in the cancer
group was high. In contrast, the amount of WBC decreased dramatically in mice treated
with BVFCE, and doxorubicin compared to the mice group without treatment, which
showed a high number of leukaemic cells. Liver, kidney, heart and spleen were stained
with haematoxylin and eosin (H&E) and viewed under a light microscope. Spleen and
liver weights were significantly low (p< 0.05) in the mice treated groups compared to
the untreated group, indicating a significant splenomegaly and hepatomegaly. From the
results above, it can be seen that the BVFCE has high anti-leukaemic activity, which
might be due to its antioxidant. In conclusion, daily intake of this fruit by groups
suffering from leukaemia could suppress or decrease cancer development with no or
fewer side effect than the normal chemotherapeutic drug. |
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