Rationale

Diabetes mellitus is a complex disorder that is characterized by hyperglycemia resulting

from malfunction in insulin secretion and/or insulin action both caused by impaired metabolism

of glucose, lipids and protein (Scheen, 1997). The chronic hyperglycemia of diabetes is

associated with long term damage, dysfunction and failure of various organs (Lyra et al., 2006).

In diabetic rats, the utilization of impaired carbohydrate leads to accelerated lipolysis, resulting

in hyperlipidemia (Morel and Chisolm, 1989; Murray, 1996).

Despite the presence of known anti-diabetic medicine in the pharmaceutical market,

diabetes and the related complications continued to be a major medical problem. Recently, some

medicinal plants have been reported to be useful in diabetes worldwide and have been used

empirically as anti-diabetic and anti-hyperlipidemic remedies (Mitra et al., 1996; Shukla et al.,

2000; Bhattaram et al., 2002; Mahomed and Ojewole, 2003; Hou et al., 2005; Huang et al.,

2005).

Herbal medicine development against the non-communicable disease like diabetes is one

of the thrust area of research in the field of global medicine. Many traditional medicines in use

are derived from medicinal plants, minerals and organic matter. The World Health Organization

(WHO) has listed 21,000 plants, which are used for medicinal purposes around the world.

In the Philippines, Vaughn Alviar a journalist of the Philippine Daily Inquirer stated in

his article last March 2016 that the Philippines is now a diabetes hotspot. Philippines is ranked in

the top 15 in the world for diabetes prevalence, it being is home to more than 4 million people

diagnosed with the disease – and a worryingly large unknown number who are unaware they

have diabetes.(International Diabetes Federation).

 Diabetes mellitus is known as one of the most epidemic disease in the Philippines.

Locally, diabetes ranked 5th among the top ten causes of death in the Philippines (Department of

Health, 2016). The mortality rate caused by diabetes mellitus is largely related to the

complications developed by persons affected by this disease. Yu-Gan (2013) related that diabetes

is considered as the primary cause of cases of amputations, stroke, blindness and kidney failures

in the Philippines. The cases of death in the Philippines due to diabetes is parallel with the study

by Mathers and Loncar (2006). It states that about 80% of the people who die from

complications of diabetes belongs to low- and middle-income countries, i.e. developing countries

such as the Philippines.

Pinto peanut or perennial peanut (Arachis pintoi Krap. & Greg.), locally known as mani-

manian in the Philippines, may be found growing along roadsides, and has now become a

popular landscape ground cover. It also shows some drought resistance but not too waterlogged

and poorly structured clay. In addition, it tolerates soils with 70% or greater aluminum (Al)

saturation (Pinto et al. 2004; Cook 1992). Establishment of the plant can either be through the

seed, cuttings or stolons. Mani-manian is primarily used as a cover crop in coffee, banana, citrus,

pineapples and other crops and as a ground cover that usually gives aesthetic value.

However, no scientific investigation has so far been conducted on the anti-diabetic

activity of mani-manian. The present work was therefore undertaken to study the glucose-

lowering effects of of mani-manian leaves in alloxan-induced albino mice.

Research Questions

1. Does the Mani-manian leaves extract have the potential to decrease the blood sugar level

of the alloxan-induced albino rats?

2. Is there a difference on the blood sugar level of the alloxan-induced albino rats between

before and after the administration of the following?

- Metformin (Positive Control)

- Mani-Manian Extract

- No Treatment (Negative Control)

3. Will there be differences on the blood sugar levels of the alloxan-induced albino rats after

the administration of the different treatments on the following intervals?

a) 1st hour

b) 2nd hour

c) 3rd hour

4. Is the Mani-manian leaves extract better than the commercially purchased drug

(metformin) in terms of its glucose-lowering effect?

Hypothesis

Null Hypothesis

1. The Mani-manian leaves extract have no potential in decreasing blood sugar level of the

alloxan-induced albino rats.

2. There are no differences on the blood sugar level of the alloxan-induced albino rats

between before and after the administration of the following:

- Metformin (Positive Control)

- Mani-Manian Extract

- No Treatment (Negative Control)

3. There are no differences on the blood sugar levels of the alloxan-induced albino rats after

the administration of the different treatments on the following intervals:

a) 1st hour

b) 2nd hour

c) 3rd hour

4. The glucose-lowering effect of the commercially-purchased drug (metformin) has more

potential than the Mani-manian leaves extract.

Expected Outcomes

The objective of the study is to prove the glucose-lowering effects of Mani-manian leaves

in alloxan-induced albino mice. Specifically, this study aims to determine the decrease of their

blood sugar level in every hour in three hours after the administration of the test drug.
Procedures

Research Design

The research study used experimental method to know if Mani-Manian (Arachis pintoi)

leaves extract glucose-lowering effect on Alloxan-induced Diabetic Rats. The design that was

used in the study is the experimental research design particularly the completely randomized

design. Methodologies of other research studies were used and adopted in this study.

Plant Extraction

300 grams of Mani Manian (Arachis pintoi) leaves were boiled in 900mL of water for

fifteen (15) minutes. The mixture was filtered with cheesecloth to separate the liquid part from

the solid part.

Preparation of Diabetic Mice

The test animals were acclimatized for one week prior to the administration of the

different doses of Alloxan monohydrate. Before the administration of Alloxan, the blood glucose

level was acquired through the use of tail cut technique and was read with Accu-Check

glucometer for their initial blood glucose level.

500mg of alloxan monohydrate were dissolved on 25mL Natural Saline Solution (NSS).

The overnight fasted rats were induced to diabetes mellitus by a single dose of alloxan (120

mg/kg) was injected intraperitoneally (Stanely, Prince & Menon, 2000; Madhavan et al., 2008;

Venkatesh et al., 2010; Antido et al., 2017).

 1 gram of glucose was dissolved 20mL water. 2 days after the injection of alloxan, 1mL

of glucose were orally given. The blood sample was acquired 30 minutes after the administration

of glucose.

Hyperglycemia was certified in the rats by measuring fasting blood glucose two days

following alloxan injection (Ragavan & Krishnakumari, 2006; Rajagopal & Sasikala, 2008;

Antido et al., 2017). An increase of their blood sugar levels is considered diabetic and is used for

the study.

Administration of the Test Drug and Control

The test animals were immediately subjected to the administration of test drug and

control after measuring their blood sugar level. The leaf extract and the control were

administered orally once, effects on blood glucose levels were measured every after 1 hour in 3

hours.

The test animals were divided into 3 treatments with 4 replicates. The administration of

drug or control was followed as indicated in the table.

Table 1. Categorization of Alloxan-induced albino rats after the administration of Glucose.

Group Administration
I Negative Control (No Treatment)
II Positive Control (Treated with Metformin)
III Treated with 1mL of Mani-manian extract
Collection of Data

Blood samples were collected by cutting the tail-tip of the rats. The blood glucose levels

of the animals were determined after 30 minutes of the administration of glucose, and the 1st

hour, 2nd hour and 3rd hour upon the administration of treatments. This will serve as the data that

will be used in the study.

Data analysis

Gathered data was represented through graphical presentations to identify the glucose-

lowering effect of the test drugs. Decreasing curve resulted from the graph is considered an

effective glucose-lowering drug.

For the comparison of the glucose-lowering effect between Mani-manian leaves extract

and the commercially purchased drug (Metformin), the researcher compared the number of the

decreased blood sugar level of the albino rats every after hour in 3 hours. The greater decrease of

blood-glucose level of the test drug every hour is considered and concluded as more effective in

terms of anti-hyperglycemic property.