Name: Noreen Mitzi L.

Lopez Date: July 5, 2022

Title of the Study: Anti-Diabetic Activity of Diplazium esculentum (Fiddlehead Fern) Plant Extracts In Reducing the Blood Sugar Levels of
Home Based Mouse (Mus musculus)
Exercise 6: Proposed Tables for Every Objective
Objective 1. To determine if fiddlehead fern (Diplazium esculentum) has effects on reducing the blood sugar levels of home based mouse (Mus
musculus);
Methodological approach No. of Treatments No. of Control Group Data Collection

Induction of Diabetes in Group 4 - Diabetic + VS Group 1 – Normal Control To find out if a treatment
Experimental Animals. works, researcher are
Methanolic Ext (250 mg/kg) Group 2 – Diabetic Control
randomly assigned to a
Using the method of (Graham et al., Group 5 - Diabetic + VS Group 3 – Glibenclamide Control treatment or control group. In
2011) Diabetes will be induced in the treatment group, they get
mouse by a single intraperitoneal Methanolic Ext (500 mg/kg)
the treatment that they
injection of streptozotocin (140-160 Group 6 - Diabetic + VS supposed to be, and in the
mg/kg it depends) in overnight control group, they don't get it
fasted mouse weighing 10-15 g. The Aqueous Ext (250 mg/kg)
the usual and specific solution.
researcher followed the dose-range Group 7 - Diabetic + VS
recommendations of the Clinical
Islet Transplant (CIT) consortium Aqueous Ext (500 mg/kg)
regarding procedures for the mouse
bioassay in testing human islet cell
preparations to be used in clinical
trials (150 to 240 mg/kg). The
presence of hyperglycemia will
confirmed if the presence of elevated
glucose levels after 72 hours later.
Diabetic animals will be defined as
those with a blood glucose level
greater than 150 mg/dl.
Objective 2. To evaluate what anti-diabetic biological properties can be found in fiddlehead fern (Diplazium esculentum) extracts;
Methodological approach List of Biological/Chemical Compound Data Collection

Experimental Design  Flavonoids By conducting experiments using known

materials or processes, the chemical or
Phytochemical studies on leaves and in vitro  High phenolics
biological properties of a substance can be
callus, preparation of folk plants and leaf  Alkaloids evaluated. If a material has a certain effect on
extracts, methanolic and aqueous preparation a substance, then the substance has a particular
and induction of diabetes.  Glycosides
property. Additional qualities can be deduced
 Tannins when a method alters the substance. The more
Using the method of (Tomar and Sisodia, experiments are carried out, the more quality
2012) the mouse will be divided into seven  Terpenoids
may be determined. As time goes on, the
groups of six each. For 15 days, the extracts  Steroids properties that were found during the
will be administered. Group I: Normal control experiment can be used to clearly identify the
mouse will try to give saline; Group II:  Carbohydrates
drug as well as all of its known properties.
Diabetic control mouse administered saline;  Fats
Group III: Mouse will be given glibenclamide
(2.5mg/kg) daily for 15 days; Group IV:
Diabetic mouse will be given VS methanolic
extract around (250 mg/kg); Group V:
Diabetic mouse will be given VS methanolic
extract (500 mg/kg); Group VI: Diabetic
mouse will be given VS Aqueous extract (250
mg/kg); Group VII: Diabetic mouse will be
given VS aqueous extract (500 mg/kg). On
days 0, 5, 10, and 15 following extract
administration, fasting blood glucose will be
calculated.
Objective 3. To test the efficacy, safety, and toxicity of fiddlehead fern (Diplazium esculentum) as an alternative medicine for reducing the blood
sugar level of home based mouse (Mus musculus);

Methodological approach No. of time/days spent observing lethality Data Collection

Acute Toxicity Assessment  After 24 hrs The goal of acute toxicity studies is to identify
the dose that, whether administered once or
Excellent condition of mouse of either sex were  72 hours
over several administrations, will result in
deprived overnight and placed into two groups
 14 days fatality or major toxicological consequences.
of five mouse each. The extracts will orally
They also provide details on the doses that
administered at escalating doses of 100, 500,
ought to be applied in future research.
1000, 3000, and 5000 mg/kg body weight
(Barik et al., 2008). According to OECD
recommendations, the study of acute toxicity
was carried out. For a period of two hours, the
mouse should monitored continually under the
following profiles of (Barik et al., 2008). (I)
Behavior pattern. A state of alertness,
restlessness, irritation, and terror. (II)
Neurological pattern. Activities that occur
spontaneously, responsiveness, touch reaction,
pain response, and gait. (III) Profile of
autonomic function. The act of defecating and
urinating. They were observed for lethality or
death after 24 hours, 72 hours, and 14 days.
Objective 4. To evaluate which extracts have the potential to lower the blood sugar level of diabetic mouse (Mus musculus);
Methodological approach Effects of Aqueous Extracts Effects of Methanolic Extracts Data Collection
(Checklist, Yes or No) (Checklist, Yes or No)

Evaluation of potential extracts on  From 0 to 15 days,  From 0 to 15 days,does In an experiment, the

lowering mouse blood sugar levels researcher chooses a course of
does the blood sugar the blood sugar level of action and monitors the
The method of (Shirwaikar et al.,
2006) was imitate to test the oral level of mouse mouse gradually drop outcome. To compare and
glucose tolerance of normal mouse evaluate the efficacy of a
gradually drop using using methanolic treatment, a control group (a
that had been fasted overnight (18
h). Six groups of six mouse each aqueous extracts? extracts? group getting no treatment or
were given 0.9 percent (w/v) saline, a placebo) may occasionally
 Does the blood sugar  Does the blood sugar be utilized.
2.5 mg/kg glibenclamide, and
Diplazium esculentum methanol level of those who are level of those who are
and aqueous extracts 250 and 500
normaglycaemic normalgylcaemic
mg/kg respectively. After that
glucose (3 g/kg) will fed 30 minutes mouse drops mouse drops
after the extracts were
significantly using D. significantly using D.
administered. At 0, 30, 60, and 120
minutes after orally administered to esculemtum aqueous esculenetum
the experimental mouse using a
extract? methanolic extracts?
force feeding syringe, blood
samples will be collected for the
measurement of blood glucose by
puncturing the lateral tail vein. Blood
glucose will be measured by using a
one-touch electronic glucometer
using a glucose test strip monitoring
meter kit.
Using the method of (Graham et al.,
2011) Diabetes will be induced in
mouse by a single intraperitoneal
injection of streptozotocin (140-160
mg/kg it depends) in overnight
fasted mouse weighing 10-15 g.
Then the mouse will be divided into
seven groups of six each. For 15
days, the extracts will be
administered. Group I: Normal
control mouse will try to give saline;
Group II: Diabetic control mouse
administered saline; Group III:
Mouse will be given glibenclamide
(2.5 mg/kg) daily for 15 days; Group
IV: Diabetic mouse will be given VS
methanolic extract (250 mg/kg);
Group V: Diabetic mouse will be
given VS methanolic extract (500
mg/kg); Group VI: Diabetic mouse
will be given VS Aqueous extract
(250 mg/kg); Group VII: Diabetic
mouse will be given VS aqueous
extract (500 mg/kg).