Microsoft word - 2008 dec 22 fulvimed sol 396 - 401 report _2_.doc

Anti-diabetic assessment of Solutions #396 to #401


Testing Institution
Diabetes Discovery Platform, South African Medical Research Council,
Tygerberg, 7505, South Africa.


22 December 2008


Testing Institution
Diabetes Discovery Platform, South African Medical Research Council,
Tygerberg, 7505, South Africa.

OBJECTIVE
To investigate the in vitro glucose uptake by C2C12 muscle and Chang liver
cells fol owing exposure to solution # 396 - #401 samples supplied by Fulvimed.
Solution samples tested
¾ Sol 396 [concentration 5.3%] ¾ Sol 397 [concentration 6.8%] ¾ Sol 398 [concentration 8% ] ¾ Sol 399 [concentration 3.2%] ¾ Sol 400 [concentration 4% ] ¾ Sol 401 [concentration 5.2%]
In vitro Cell Culture Models

Solutions were tested in a C2C12 muscle cell in vitro glucose uptake model
For glucose uptake experiments cells were exposed to the relevant solutions at a
concentration of 0.05 µg/µl, metformin or insulin at concentration of 1 µM in
serum-free media supplemented with 8 mM glucose for one hour. Thereafter, the
glucose concentration in the media was determined by a colourometric glucose
oxidase method (Biovision Inc, USA).
Solutions were tested in a Chang liver cell in vitro glucose uptake model
Solutions were added to fresh culture media at a concentration of 0.05 µg/µl and
cultured on Chang cells for three days prior to performing the glucose uptake
experiments. As positive controls metformin or insulin were added to the media
at concentration of 1 µM. Acute glucose uptake was performed after exposing
the Chang cells to fresh serum-free media with 8 mM glucose, supplemented
with either metformin, insulin as controls and the relevant solutions for three (3)
hours. Thereafter, the glucose concentration in the media was determined by a
colourometric glucose oxidase method (Biovision Inc, USA).

Results:

C2C12 muscle cell glucose uptake

C2C12 muscle cell glucose uptake %
The above graph shows the percentage glucose uptake from the culture media supplemented with 8 mM glucose by C2C12 cells over one (1) hour.
C2C12 muscle cell glucose uptake data

Glucose conc.
Glucose Uptake
(nM/ul) SD (nM/ul) %
increase
Metformin
The above table shows the glucose uptake data of C2C12 cells following 1 hour exposure with the relevant solutions in serum free media containing 8 mM glucose. The glucose concentration column represents the glucose
concentration remaining in the media fol owing one (1) hour exposure to the cells.
The glucose uptake column represents the glucose uptake from the media during
the 1 hour exposure. SD represents the standard deviation. The percentage
increases calculated from the relevant solvent vehicle and the P= values are
reflected in the last two columns respectively.
Chang liver cell glucose uptake

Chang liver cell glucose uptake %
ose uptake %
c
lu
G
The above graph shows the percentage glucose uptake from the culture media supplemented with 8 mM glucose by Chang cells over a three (3) hour period. Chang cells were pre-exposed to the solutions tested for 3 days prior to the glucose uptake experiments. Chang Liver Cell: Glucose Uptake Data

Glucose conc.
Glucose Uptake
(nM/ul) SD (nM/ul) %
increase
Metformin

The above table shows the glucose uptake data of Chang liver cells fol owing 3
days of pre-sensitization with the relevant solutions, followed by a 3 hour glucose
uptake assay with media containing 8 mM glucose. The glucose concentration
column represents the glucose concentration remaining in the media following
three (3) hour exposure to the cells. The glucose uptake column represents
glucose uptake from the media during a 3 hour exposure. SD represents the
standard deviation. The percentage increases calculated from the relevant
solvent vehicle and the P= values are reflected in the last two columns
respectively.


Discussion of Results

Solutions 398, 399, 400 and 401 induced statistically significant increases in the
glucose uptake of the C2C12 muscle cel s in culture. The increase in the glucose uptake was comparable to that of metformin an established anti-diabetic agent. In the Chang liver cells solutions 397, 398, 400 and 401 induced statistically significant increases in the glucose uptake (p≤0.0005). The respective solutions induced an increase in glucose uptake similar to that achieved by insulin or Conclusion
• The solutions tested significantly enhanced glucose uptake both in the

Source: http://www.fulvihealth.co.za/pdf/diabetes/diabetes_in_vivo.pdf