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ANTI DIABETIC ACTIVITY OF ETHANOLIC LEAVES EXTRACT

OF ANDROGRAPHIS PANICULATA ON ALLOXAN INDUCE
DIABETIC IN RAT
Binita Dutta*1, Md Kabirul Islam Mollah*2, Snehali Banerjee*3,
Rakiba Sultana*4, Anjusha Pahari*5
*1,2,3,4,5 Department Of Pharmacology, Bharat Technology, Uluberia, Howrah, West Bengal, India,
711316
ABSTRACT
Aims: The anti-diabetic effect of ethanolic leaves extract of Andrographis paniculata was inspected on Albino
rats. Place and Duration of study was carried out between October 2020- January 2021 in the pharmacology
Laboratory at Bharat Technology, Uluberia, Howrah, West Bengal, India. Methodology: Diabetic condition was
introduced with an intraperitoneal injection of 120mg/kg BW alloxan monohydrate in the rats. After stable
diabetic condition, the rats were administered with ethanolic extract of Andrographis paniculata for 10
consecutive days. Blood glucose profiles in both preprandial and postprandial were monitored at the day of 0,
3, 7and 10. In the study, the diabetes of the rat may decrease after administration of the herbal extract of A.
paniculata. The extract may display good hypoglycemic effects. In conclusion, the ethanolic extract of A.
paniculata is potential to improve as an antidiabetic agent.
Keywords: Andrographis Paniculata Leaves, Andrographiloid, Diabetes, Alloxan Induce Diabetes.

I. INTRODUCTION
Diabetes is a disease in which the body’s respond to the hormone insulin is impaired, causing abnormal
metabolism of carbohydrates as a result of which glucose level elevated in the blood. Three types of diabetes
are there:
(1) Type 1- It is chronic medical condition where the pancreas produces small or no insulin
(2) Type 2- It is chronic medical condition that the body becomes resistant to insulin
(3) Gestational diabetes - This condition is due to insulin-blocking hormones produced during pregnancy. [1]
According to the IDF (International Diabetes Federation) report, India, china and United states have higher
rates of most cases of Diabetes. Around 470 million adults (18-80 years) worldwide were living with Diabetes.
The total number of diabetes patients is likely to be more than double in 2040. [2]
Alloxan which is an organic compound also known as 5, 5-dihydroxyl pyrimi-dine-2, 4, 6-trione, a urea
derivative, cytotoxic and carcinogenic glucose analogue. The molecular formula of this compound is C4H2N2O4
and a relative molecular mass of Alloxan is used to introduced diabetes in mice. [3, 4] Kalmegh (Andrographis
paniculata) is commonly known as “king of bitter” and it is belonging in family Acanthaceae. It is used to treat a
variety of infectious and chronic diseases as well as Diabetes. A. paniculata is one of the most commonly used
medicinal plants and it is found in part of Asia and European countries. This local plant that are potentially
developed as antidiabetic agents. [5] This study was aimed to explore antidiabetic effect of herbal extract of A.
paniculate in alloxan-induced mice. Diabetic condition was introduced with an intraperitoneal injection of
120mg/kgBW alloxan monohydrate in the mice. After constant diabetic condition, the mice were administered
with the extract for 10 consecutive days. Andrographis paniculata is known for its curative and preventive
properties as whole plant is used such as stem, roots and leaves, which is the source of several diterpenoids of
which andrographolide is important constituent which shows the Antidiabetic activity. Recent experiments
have also shown that it’s has some antibiotic and antityphoid activity. The aerial part and leaves of plant are
used in treatment of diabetes, jaundice, liver disease, fever, diabetes, snake bite, dysentery, sore throat and
chronic malaria. [4]
In India, the whole plant is used to acquire andrographolide, the most important pharmacologically active
compound. Having such a broad geographical distribution and important medicinal value of this plant all over
the country, its undiscriminating collection from wild sources without compensating any attention towards its

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domestication and conservation in steady agriculture has caused a sharp drop in the availability of drug to the
industries and acceleration in its prices. Therefore, the dense demand of phytochemicals especially diterpene
lactone like andrographolide in India as well as international markets has encouraged Indian farmers to begin
commercial cultivation of kalmegh. [5, 6]
II. MATERIALS AND METHOD
Plant material collection
The sample were collected from rural belt of Purba Medinipur, West Bengal, India in the month of September
2020. After collection of Plants, it is authentified by Pharmacognosist Dr. Sanjit Das, Asst. Professor,
Department of Pharmacognosy, Bharat Technology, Banitabla, Uluberia, Howrah, West Bengal, India. The plant
was separated from undesirable materials, cleaned, washed with distilled water and shade dried in room
temperature and then powdered.
Preparation of extract
The shade dried leaves were powdered using a mechanical grinder and passed through 40 mesh sieves. Powder
300gm of powdered of leaves was successively extracted with 1.5 L of petroleum ether, chloroform and ethanol,
in a soxhlet apparatus at 60–70°C each for 10–12 h consecutively. Solvents used were of analytical grade.
Ethanol was removed from the extract by simple distillation and Evaporated to dryness and a semisolid mass
was obtained. The semisolid mass was collected carefully sand packed in eppendorf, stored in refrigerator until
use. Now the extracts were subjected to preclinical screening.
Experimental animal
Adult male and female, non-pregnant albino rats (average weighing 160 to 250 gm respectively) were used in
this study. The animals were obtained from the department of pharmacology of Bharat Technology, Uluberia,
Howrah, West Bengal, India. They were maintained under standard environmental condition (24 ± 2 °C, 60 to
70 % RH) in animal house approved by the committee for the purpose of control and supervision on
experiments on animals (CPCSEA) and fed with commercial pellet diet and water and libitum. They were taken
out to the laboratory environment for at least two weeks before experiment. The Institution of animal ethics
committee, Bharat Technology, Uluberia, Howrah, West Bengal, India approved the experiment protocol and
following the guidelines and procedures of the” Principle of laboratory animal care” (National Institute of
Health- NH publication number 85-23). [1, 7, 8]
Experimental Protocol
The animals will randomly be assigned into four groups of 6 in each group and received the treatments:
Group I: Normal control rat will be treated with vehicle alone.
Group II: Diabetic control rat will be treated with Alloxan. (120 mg/kg i.p)
Group III: Diabetic rat will be treated with Glimipride. (5mg/kg i.o)
Group IV: Diabetic rat will be treated with Andrographis paniculata extract. (400 mg/kg i.o)
All the 3 groups except normal control were injected with alloxan and were fasted for 18 hours. Then standard
drug and A. paniculata extract were given orally to the rats group at the interval of 3, 7, 10 days. Check the
blood sugar level of the rats at 2 days interval. [1]
Statistical analysis of data
The data obtained were examined by using the Graph pad Prism, version 9.1.1. All the values were presented in
the table, they expressed as mean ± standard error mean (SEM) of six animals. The important difference
between the mean diabetic index of treated group and that of the control group was tested with one-way
analysis of variance (ANOVA) followed by Dunnett’s post-test and p values <0.05 deliberated significant.[8]
III. RESULTS
Phytochemical screening
Analysing of various chemical compounds within the extract, represents the preliminary phytochemical studies.
Minor quantity of ethanolic extracts of Andrographis paniculata were subjected to preliminary quantitative
phytochemical investigation for detection of phytochemicals like Tannins, Flavonoid, Alkaloid & Glycosides,
Phenolic compounds & Tannins, Saponins, Steroid, Terpenoids. [8, 9,]

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Pharmacological study
A. Effect of administration of Alloxan (120mg/kg; i.p.) in rat.
The glucose level of Alloxan treated rat in “0” days (before induced), “3” days (after induced) “7” days and “10”
days were found to 89± 2.83, 210± 6.37, 270± 3.40, 274± 1.78. Thus, Alloxan increases the level of glucose
against vehicle treated (group I).
B. Effect of administration of Glimipride (5mg/kg orally) in Alloxan induced diabetic rat.
The glucose level of Glimipride treated rat in ‘0’ days (before induced), ‘3’ days (after induced), ‘7’ days and ‘10’
days are found to 92± 1.86, 190± 13.26, 154± 2.31, 98± 2.90. Thus, Glimipride significantly decrease the levels
of glucose against Alloxan induced (Group II).
C. Effect of administration of ethanoilc extract of leaves of Andrographis paniculata (400mg/kg orally)
in Alloxan induced diabetic rat.
The glucose level of Andrographis paniculata treated rat in ‘0’ days (before induced) ‘3’ days (after induced), ‘7’
days and ‘10’days were found 86± 2.13, 182± 3.86, 110± 2.49, 81± 2.26. Thus, Andrographis paniculata
significantly decrease the level of glucose against Alloxan induced (Group II)
Table -1: Effect of ethanolic extract of the leaves of Andrographis paniculata on Alloxan induced diabetic rat
Group Treatment ‘0’ days ‘3’ days ‘7’ days ‘10’ days

I Normal control 86±1.96 88± 2.01 90± 1.96 87 ± 1.47

Alloxan
II (120mg/kg ip) 89± 2.83 210±6.37 270± 3.40 274±1.78

Standard
III (Alloxan +Glimipride) 92 ±1.86 190±13.26 154± 2.31 98± 2.90
(5mg/kg i.o)

Test
IV (Alloxan + A.paniculata) 86± 2.13 182± 3.86 110±2.49 81± 2.26
(400mg/kg i.o)
Values are expressed as Mean SEM; (n=5); Significance relative to control: ***p< 0.001

Figure -1: Effect of ethanolic extract of leaves of Andrographis paniculata in Alloxan induced rat at ‘0’days

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Figure -2: Effect of ethanolic extract of leaves of Andrographis paniculata on Alloxan induced rat ‘3’ days

300
Blood glucose level(mg/dl)

250
*
200
*
150

*
100

50

0
l l ad st
ro tro ar Te
nt on
l co c tand etic
a c
rm eti tic
s ab
ab Di
No Di ibe
D
7 days of Treatment

Figure -3: Effect of ethanolic extract of leaves of Andrographis paniculata on Alloxan induced rat ‘7’ days

Figure -4: Effect of ethanolic extract of leaves of Andrographis paniculata in Alloxan induced rat at ‘10’ days

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IV. DISCUSSION
Nearly the kalmegh plant and their parts were discovered to be wealthy in phytochemicals, for example,
terpenes, terpenoid. Terpenes are the most widely familiar terpenoids. Terpenoids containing of Andrographis
paniculata has a great anti diabetic property, which is effectually comprehended in this examination. On the
Other hand in this exploration we examine about the number of cycle were included a medication
advancement, how potency we direct a preclinical preliminary as well as clinical preliminaries. How we
acknowledge the process of isolation, separation and recognition of the medication particle, enthusiasm a dose
structure and so on and portion valuation by LD50 (intense harmfulness test).[1, 2]
Alloxan, one of the popular diabetogenic agent which is used to assess the antidiabetic capacity of the test
compound, is a urea derivative, which is likely to inhibits glucose sensor of the Beta cell, and causes a state of
insulin dependent diabetes through its ability to introduce ROS formation, as a result of which necrosis of beta
cell occur. This experiment gives us the evidence that Kalmegh [Andrographis paniculata] has potent
antidiabetic properties in it.
Carcinogeneces, mutagenesis, and teratogenesis, diabetogenic, nephrotoxic, hepatotoxic, gastric ulceration,
these all are caused by Alloxan. Alloxan is given by intraperitoneally, intravenously to laboratory rat in multiple
sub-diabetogenic doses to produced pancreatic insulities with eventual ruin the insulin secretion in the beta
cells and as a result of which diabetes mellitus occur. The frequency and brutality of lesion produced by Alloxan
in pancreas, kidney GIT and Liver, gradually increased with the time of the post treatment.
Andrographis paniculata was able to successfully decrease the level of high blood sugar level to normal range.
The compound succeeded to improve the uptake of glucose in alloxan induced rat. Kalmegh possess some other
activities such as anti inflammatory,antispasmodic, hepatoprotective and antioxidant.
The ethanolic extract of leaves of Andrographis paniculata reduce the glucose level in Alloxan induced diabetes
in rat. Hence, Andrographis paniculata extraction may real in some pathway which aggravate the diabetes
mellitus. Such this experiment indicates this Andrographis paniculata extraction possibly will stimulate the
secretion of insulin or the activity of insulin increase. [9, 10]
V. CONCLUSION
The present study exposes that the ethanolic extract of Andrographis paniculata significantly recover the effect
of diabetes in Alloxan induced diabetic rats. Thus it concluded that the ethanolic extract of leaves of
Andrographis paniculata is potential in contradiction of to diabetic.
COMPETING INTERESTS
Authors have declared that no competing interests exist
ACKNOWLEDGEMENTS
The authors are thankful to management of Bharat Technology for providing the required facilities to carry out
the research work.
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Volume:03/Issue:05/May-2021 Impact Factor- 5.354 www.irjmets.com
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