Thesis (M. Sc.(Biochemistry)) -- University of Limpopo, 2020 / Diabetes mellitus has become a global epidemic, particularly type 2 diabetes.
Obesity is one of the causes of type 2 diabetes mellitus due to its link with induced
insulin resistance. There is no cure for diabetes mellitus and, as such, it is managed
by using standard drugs which have side effects, and can be toxic, expensive and
unavailable. People have resorted to the use of medicinal plants to treat diabetes
and its complications. The aim of this study was to test the anti-obesity and anti diabetic properties of Bidens pilosa crude extract and its sub-fractions using C2C12
myoblasts and 3T3-L1 adipocytes. The crude extract and the most active sub fractions were selected for further analysis because of their ability to stimulate
glucose uptake and induction of adipogenesis.
Bidens pilosa leaves were selected for this current study. They were firstly extracted
using absolute methanol and further subjected to solvent-solvent fractionation to
obtain the n-butanol, ethyl acetate, water, hexane, chloroform and 35% water in
methanol sub-fractions. Qualitative phytochemical analysis was performed using
thin layer chromatography (TLC) and standard chemical tests. Total phenolic and
flavonoid content were determined quantitatively using a calorimetric method with
Folin-Ciocalteu’s reagent. For their antidiabetic potential, the extracts were
evaluated chromogenically and calorimetrically for antiglycation and α-amylase
inhibitory activity. The cytotoxicity of the extracts on 3T3-L1 preadipocytes and
C2C12 myotubes were determined using the MTT assay. The adipogenesis inducing
effect of the extract was tested using the adipogenesis kit.
More compounds were found on chromatograms eluted in EMW mobile phase (Ethyl
acetate: methanol: water). The extracts were shown to contain a variety of secondary
metabolites, and high phenolic and flavonoids contents. Crude, chloroform, n butanol and water sub-fractions had high antioxidant activity. Alpha amylase activity
was highly inhibited in the crude extract and all sub-fractions, with the highest
inhibitory activity observed in the crude extract and the chloroform, n-butanol and
water Sub-fractions (IC50 1.25 ± 2.5 mg/ml). The cytotoxic profiles indicated that all
extracts are non-cytotoxic at concentrations of 15.63 µg/ml. Extracts at a
concentration of 31.25 µg/ml were shown to stimulate the accumulation of
triglycerides using 3T3-L1 adipocytes. The extracts also exhibited significant (P < 0.05) glucose uptake activity. In conclusion, Bidens pilosa contains constituents
that inhibit α-amylase, antiglycation formation and modulates uptake of glucose in
3T3-L1 adipocytes. The use of B. pilosa in combination with insulin revealed the
synergistic effects in facilitating glucose uptake in both C2C12 myotubes and 3T3-
L1 adipocytes. This suggests that there might be some binding compounds found in
the plant extracts that are responsible for the stimulation of expression of several
genes that encode for proteins involved in the metabolism of glucose. However, the
use of B. pilosa, in combination with metformin, results in a decreased glucose
uptake. Bidens pilosa have the fast-acting insulin mimetic properties. Furthermore,
the plant was shown to stimulate the accumulation of triglycerides in 3T3-L1
adipocytes, signifying the plant can induce adipogenesis at 30µg/ml / South African Medical Research Council (SAMRC)
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:ul/oai:ulspace.ul.ac.za:10386/3836 |
| Date | January 2020 |
| Creators | Tolo, M. M. |
| Contributors | Mokgotho, M. P., Mampuru, L. J., Bagla, V. P., Poopedi, K. W. |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | vi, 103 leaves |
| Relation |
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