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EFFECTS OF CATECHINS ISOLATE OF UNCARIA GAMBIR ROXB ON BURN WOUND HEALING IN MALE RAT

Muhammad Yanis Musdja, Lina Elvita & Nursetyowati Rahayu,

Faculty of Medicine & Health Sciences, State Islamic University, Jakarta

Email : myanis88@gmail.comPhone +62 812 8168 1197

Bromo Conference 2018, UNAIR11-12 July, 2018, Surabaya, Indonesia

ABSTRACTBackground. Indonesia is the largest Uncaria gambir, Roxb (gambir) producing country in the world, but its benefits as a medicinal plant has not been optimally developed. Traditionally, gambir plants have been widely used by the community as wound healers. Objective: This study aims to determine the efficacy of catechin isolate as a healing burn on white rats male Sprague Dawley strain through observation of anatomical and histopathological pathology.Methods: Catechins of gambir were isolated by using partition method with ethyl acetate solvent. Rats were made to suffer burns with Akhoondinasab method, et al. A total of 30 male white rats were divided into 5 groups, given daily test preparations. A total of 3 groups were given catechin isolates prepared by gel with variations of concentrations of 1%, 2% and 4%). One group was used as a comparison with a patent medicine for healing burns and one other group was used as a negative control. After administration of the dosage for 21 days, the pathology and histology changes was observed in the burns that occurred. Results: Statistical analysis showed that gel isolate catechins of Gambir with concentrations 2% showed the effect of decreasing the burn area and an increase in the percentage of healing of burns that do not differ significantly from positive controls (P <0.05). Histopathologic observations showed that gambir catechin isolates were able to reduce the number of inflammatory cells and increase neocapillary formation compared with the negative control group. Conclusion: Catechin isolates of gambir with concentration 2 % can help the healing process of second degree of burns in the inflammatory phase and proliferation.

Key words : Uncaria gambir, Catechins isolate, burn wound healing, male rats

INTRODUCTION:

Gambier (Uncaria gambier Roxb) has since long been used by many people in the ASEAN region traditionally as anti-microbial, anti-diarrhea, and agent for the treatment of burns. Gambier is also one component for chewing medicine to strengthen teeth and antimicrobial.

O

OH

HOH

OH

(OH)

OH

(+) - catechin

Dry extract of gambir water

Methods: Gambir was obtained from Limapuluhkota districts, Indonesia, These districts are the largest gambir producer in the world. Screening of chemical compound group of gambir extract was done based on Harbone methods. Catechins isolation was done based on National Standard of Indonesia: SNI 01-3391-2000, “A total of 250 g of gambir powder was extracted with a water solvent at temperature of 90 - 96 °C for 15 minutes while stirring. Then the infusion was filtered in hot conditions using a funnel coated with filter paper. The extract obtained was partitioned with ethyl acetate, the ratio of extract with ethyl acetate (1 : ½.). Ethyl acetate phase was taken and the water phase was partitioned repeatedly with ethyl acetate until a clear solution was obtained. The ethyl acetate phase was condensed with an evaporator, then washed with cold water and filtered. Catechin that obtained was dried in an oven at temperature 70 °C.

The yield of catechins obtained from the gambirisolate was determined as (+)-catechins by comparing with standard (+)-catechins and measured by a spectrophotometer UV-VIS at wavelength 279 nm.

Quality of gambir was determined based on National Standard Indonesia: SNI 01-3391-2000.

Characteristics Requirements Gambir sample

Color Yellow to yellow brownish Yellow to yellow brownish

Smell Typical Typical

Shape Solid, irregular cube Solid, irregular cube

Flavors A little bitter ending taste

rather sweet

A little bitter ending taste rather

sweet

Water content Maximum 14% 1,07%

Ash content Maximum 5% 3,61%

Catechin levels Minimum 60% 88,65%

Results of Quality Control of The GambirRequirements According to SNI (2000) and Pharmacopoeia Herb I (2008)

Histopathology Observation Scoring is based on Asian Journal Animal and Veterinary Advances (Hosseini, 2011)

Skor Inflammatory Cells Angiogenesis

0 13-15 inflammatory cells per field of view There is no angiogenesis, There is

congestion, hemorrhagic and edema

1 10-13 Inflammatory cells per field of view 1-2 blood vessels per field of view,

edema, hemorrhagic, congestion

2 7-10 inflammatory cells per field of view 3-4 blood vessels per field of view,

edema, hemorrhagic, congestion

3 4-7 inflammatory cells per field of view 5-6 of blood vessels per field of

vision, edema, hemorrhagic,

congestion

4 1-4 inflammatory cells per field of view More than 7 blood vessels per field

of view, edema, hemorrhagic,

congestion

Results of Histopathology Scoring

Group Score

Neocapilerization Infiltration of

inflammatory cells

Positive Control 3 4

Negative Controls 0 0

Test Concentration

1%

1 2

Test Concentration

2%

2 3

Test Concentration

4%

0 1

Testing Healing of Wound on Rats caused of Effects of catechin isolates Gambir

Group Number of rats

Treatment Description

I 6 Positive control, given patent burns medicine

21 days

II 6 Negative control, given gel, without catechins

21 days

III 6 Gel with catechins isolates 1 % 21 days

IV 6 Gel with catechins isolates 2 % 21 days

V 6 Gel with catechins isolates 4 % 21 days

Giving of the gel is done topically every day for 21 days with a dose of 2 times a day in the morning and evening.

Histopathological image of white mouse skin tissue with Hematoxylin-Eosin staining at 200x and 400x magnification

CONCLUSION• Gambir catechin isolate (Uncaria gambir) in the

concentration group of 1%, 2% and 4% there was nosignificant difference to decreasing wound area andpercentage of second degree burn healing which was giventopically on anatomical pathology observation.

• There was a significant difference in infiltration ofinflammatory cells and neocapillaries on day 7 in the 2%concentration test group compared with the 1%concentration test group, 4% concentration test andnegative control on microscopic observation.

• Gambir catechin isolates can assist in the healing process ofburns in the inflammatory phase and proliferation.