114 AMJ March, 2015

Effect of Centella asiatica Leaves on Gastric Ulcer in Rats

Ester Mariska1, Trully D. Sitorus2, Januarsih A. Rachman3 1Faculty of Medicine, Universitas Padjadjaran, 2Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Padjadjaran, 3Department of Biology Cell, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia

Abstract

Background: Gastric ulcer is disruption of gastric mucosal integrity due to lack of smaller defensive factors (e.g. mucus levels) than gastric aggressive factors. Centella asiatica is a traditional medicine that contains Centella asiatica leaves for preventing gastric mucus level reduction of aspirin-induced gastric ulcer model triterpenoidsin rats and compared and flavonoids. it to misoprostol. The objective of this study was to analyze the effect of ethanol extract of Methods: aspirin 450mg/kg, An experimental group III laboratory (tested group) study usingreceived 24 ratsCentella that wereasiatica divided 400 mg/kginto 4 groups. and aspirin Group 450 I (negative mg/kg, control) received carboxymethy cellulose (CMC) solution, group II (positive control) received CMC and group IV (compared group) received misoprostol 72μg/kg and aspirin 450mg/kg for three days. Then theResults: rats were laparatomied and their gastric mucus levels were measured. The results were statistically analyzed by Independent T-test. The mean levels of gastric mucus of group I, II, III, IV, respectively were 0.112; 0.035; 0.537; 0.455 opticalgroup and densities compared per group. gram of tissue. The statistical test suggested a significant difference between the positiveConclusions: control Centella group asiaticaand tested leaves group. can preventMoreover gastric there mucus is no significantlevel reduction difference of aspirin-induced between the gastrictested ulcerKeywords: model Centella in rats and asiatica has similar, gastric effect mucus, in gastricgastric mucusulcer, misoprostol level with misoprostol.

Introduction

2 Gastric ulcer is disruption of gastric Traditional medicines The percentage have been of Indonesian used for treatment from generation to generation based mucosal integrityand arises from the 3 smaller level of defensive factors (e.g. on experiences. Traditional medicine mucus levels) than gastric aggressive ispopulation medicine who derived use traditionalfrom plants, medicine animals, is about 22.26% in 2008. One of these traditional medicines is Centella and antiplatelet over a long period of time. minerals,asiatica. or a mixture of these materials. factors.1This condition can be triggered Centella asiatica is a short-stemmed by the use of aspirin as anti-inflammatory for leprosy, syphilis, fever, and plays a role andPatients vomiting. with The gastric pain ulcers arises often after complain eating. ofThis epigastric situation pain, led burning to decreased sensation, patient’s nausea, creeping plant that have been used as a cure productivity. Therefore, the patients require in wound healing, increasing work capacity medicine, such as misoprostol, or traditional of neurotransmitters in the brain,Centella and asiatica treatmentsmedicine. The which mechanism may include of action modern of revitalizing body tissues. It grows in tropical misoprostol as a prostaglandin analogue and subtropical regions, e.g. Indonesia. is stimulating the secretion of mucus and and small has fruits. branched The height tap root, of Centella green leaves asiatica in ais fan-shaped 10-50 cm.4 or kidney-shaped, round flowers, 1 the effect of Centella asiatica against gastric bicarbonate, reducing mucosal cell turnover, The objective of this study was to analyze andCorrespondence: increasing mucosalEster Mariska, blood Faculty flow. of Medicine, Universitas Padjadjaran, Jalan Raya Bandung-Sumedang Km.21, Jatinangor, Sumedang, Indonesia, Phone: +62 813 8684 1321 Email: [email protected]

Althea Medical Journal. 2015;2(1) Ester Mariska, Trully D. Sitorus, Januarsih A. Rachman: Effect of Centella asiatica Leaves on Gastric Ulcer in Rats 115

Figure 1 Procedures of The Experiment ulcers and to compare the effect of Centella asiatica 24 male wistar strain rats with inclusion Methodswith misoprostol. criterias as follows: 8-12 weeks old, 150-200 grams weight, active movement, clean hair, no wound on the body, and normal stools. The exclusion criteria are weight lost more than AnAlthea experimental Medical Journal. study 2015;2(1) was carried out, using 10% of the initial body weight during the 116 AMJ March, 2015

adaptation and sickness after the adaptation. of Sekolah Ilmu Teknologi Hayati Institut repair, as well as stimulating mucosal damage. The rats were obtained from the Laboratory Theif NSAIDs NSAIDs are arein acidic weak environment. acids. The NSAIDsIn that will remain as non-ionized lipophilic form Teknologiof Dr.Hasan BandungSadikin General and wereHospital adapted Bandung for seven days in the Pharmacological Laboratory condition,in the cell. NSAIDsIf the drugs will crossare trapped through inside the lipid the environment.5 The leaves of Centella asiatica membranes of epithelial cells and be1The trapped dose to prevent or minimize the stress in a new cell, then the cell will be damaged. were obtained from Kebun Percobaan Manoko, of aspirin was 450 mg/kg body weight. The Kecamatan Lembang, West Bandung, West dosesand humans.used in this7 study were the results from Java and extracted in Balai Penelitian Tanaman conversion using table experimental animals Rempah dan Obat, Bogor, West Java. The fresh ml CMC solution and Centella given orally asiatica to the extracts, rats.8 leavesfrom dirt were or harvested foreign material at the age and of reduce 3-6 months. from aspirin, and misoprostol were dissolved in 5 The leaves were sorted and washed to clean celluloseThe rats (CMC) were dividedsolution, into group 4 groups. II (positive Group microbial. Then, the leaves were pulverized. Icontrol) (negative received control) CMC received and aspirin carboxymethy 450mg/ Next, the sample was mixed with ethanol kg, group III (tested group) received Centella (70%) with a ratio of 1:5 in shaking water asiatica 400 mg/kg and aspirin 450 mg/ bathconcentrated for 3 hours in a and rotatory allowed evaporator to steady at for 500C 16 kg, group IV (compared group) received hours. The sample was filtered by filter paper, for 6 hours, and dried with a freezer dryer for misoprostolto determine 72μg/kgthe normal and gastric aspirin mucus 450mg/kg level in 2 days. Twenty kg of wet Centella asiatica plant during three days. The function of group I was (all parts) became 836.5 grams of dried leaves6 and became 96 grams of dried extract. The rats. After 3 days, all the rats were scarified by dose of extract was 400 mg/kg body weight. ketamine and were laparatomied. Their gastric Misoprostol was used as a comparative drug mucususing the levels Green were AP, collectedLander JE, and Turner, measured DH andanalogue obtained has from mechanisms PT Novell Pharmaceuticalof action for by spectrophotometer (wavelength 605 nm) Laboratories.stimulating the Misoprostol secretion as of a prostaglandinmucus and (1981)Unit Penelitian method. Kesehatan The measurement Rumah Sakit of gastricUmum 1 The dose mucusPusat Dr. levels Hasan was Sadikin done Bandung. in the Laboratory The results of bicarbonate, reducing mucosal cell turnover, and increasing mucosal blood flow. of misoprostol was 72 μg/kg body weight. In were analyzed using Independent T-test. The this study, Aspirin was used to create a gastric test in this study used Confidence interval (CI) ulcer condition in the rats and was obtained 95% (α= 5%). The measurement from(NSAIDs). PT Bayer Mechanism Indonesia. of Aspirin action belongsof NSAIDs to a The procedures were based9 on the research class of Nonsteroidal Anti-inflammatory Drugs of M. Sancar et. al. (2009). 10 (Figure mechanism can harm mucosal defense and of1) mucus levels were based on the Green AP, is to inhibit synthesis of prostaglandin. This Lander JE, TurnerDH (1981) method.

Table 1 Gastric Mucus Levels of the Gastric Ulcer Induced Rats Gastric Mucus Level (optical density units per gram of tissue) Group Group I Group II Group III Group IV 1 0.186 0.047 0.585 0.528 2 0.050 0.028 0.720 0.370 3 0.070 0.061 0.652 0.573 4 0.140 0.015 0.361 5 0.134 0.011 0.411 0.556 0.339 6 0.048 0.513 0.342 Mean(SD) 0.112(0.051) 0.035(0.020) 0.537(0.145) 0.455(0.108) 0.092

Althea Medical Journal. 2015;2(1) Ester Mariska, Trully D. Sitorus, Januarsih A. Rachman: Effect of Centella asiatica Leaves on Gastric Ulcer in Rats 117

Table 2 Effect of Aspirin in Reducing Gastric Mucus Levels in Rats n Mean (SD) Mean Difference (CI 95%) P* Group I 6 0.112 (0.051) 0.077 (0.024-0.130) 0.006 Group II 6 (0.035 0.020)

prevent declining of mucus levels of aspirin- Results induced gastric ulcer model in rats (p<0.05). Centella asiatica consists of triterpenoids The results of measurement of rat gastric common component in Centella asiatica.4 and flavonoids. Triterpenoid is the most mucus levels can be seen in Table 1. et al. There was a significant difference in the Centella asiatica has been proved to have induction of aspirin (450 mg/kg) can decrease rolesstated asthat wound asiaticoside healing, derived anti-inflammatory, from Centella mean of mucus level of gastric between asiatica groupmucus I leveland IIand (Table create 2). Thisa gastric result ulcer states model that and antioxidant. A research of Shukla in rats. had a role in wound healing in vitro and in vivo. It was proved by increasing tensile strength, collagen, fibroblast proliferation, and There was a significant difference in the the increasing angiogenesis in vitro.12 The mean of mucus level of gastric between wound epithelization in guinea pig and rat. Its declining of mucus levels of aspirin-induced researchrole in wound also stated healing that was saponin, also proved asiatic fromacid, group II and III (Table 3). This result state madecassoside derived from Centella asiatica thatgastric Centella ulcer model asiatica in rats. extract can prevent a the production of collagen type I to stimulate 4 There was no significant difference in the had the effect of wound healing by inducing mean of mucus level of gastric between group preventing gastric mucus level reduction of the wound healing process. III and IV (Table 4). This result state that and remodeling. Centella asiatica has a role Centellaaspirin-induced asiatica gastric extract ulcer has similar model effectin rats in Normal phases of wound healing consist ofin the hemostasis, proliferative inflammation, phase. This proliferation, phase is withDiscussions misoprostol. dominated by granulation tissue formation andfrom epithelization. platelets to Instimulate this phase migration there will and be cells on the surface of gastroduodenal releasing of macrophage and growth factor The gastric mucus is produced by epithelial 13 mucin, and lipid. The functions of gastric activating of fibroblast. Then, fibroblast will produceResearch an of important Frederico substance et al. stated for that wound there organ. The mucus consists of water (95%), healing. One of the substances is collagen. mechanical damage, protect gastric lining received from Centella asiatica mucus are lubricate to protect mucous from isCentella a positive asiatica correlation between the flavonoids radicals. Free radicals can damage and antioxidant. proteins, from self-digestion from pepsin (without 14 1,11 can absorb or neutralize Research free disturbingthe function the activityof HCl ofin pepsin) the lumen). and from of Huang et al. stated that asiatic acid derived acid by neutralizing HCl (without disturbing from Centella asiatica had the effect as anti- Centella asiatica DNA, and lipids in cell membrane. to administrationThis study showed of aspirin that rats (450 which mg/kg) received for three consecutive extractdays had (400 mean mg/kg) mucus priorlevel 0.537 optical density per gram of tissue. It inflammatory. Anti-inflammatory response can demonstrated that Centella asiatica be seen from declining malondyaldehyde and nitric oxide levels. Both of them are produced by impact of free radicals that attack plasma Table 3 Effect of Centella asiatica extractand Aspirin-induced can Gastric Ulcer in Gastric Mucus Level n Mean (SD) Mean Difference (CI 95%) p* Group II 6 0.035 (0.020) 0.502 (0.350-0.653) <0.001 Group III 6 0.537 (0.145)

Althea Medical Journal. 2015;2(1) 118 AMJ March, 2015

Table 4 Effect of Centella asiatica and Misoprostol in Gastric Mucus Level Induced rats n Mean (SD) Mean Difference (CI 95%) p* Group III 6 0.537 ± 0.145 0.081 (0.083-0.246) 0.147 Group IV 6 0.455 ± 0.108

editors. Evaluation of drug activities: membrane. Anti-inflammatory response In: Lawerence DR, Bacharach AL, can also be seen by increasing antioxidant enzyme,tissue.15 A e.g. research catalase, of Somchit superoxide et al. stated dismutase that 8. pharmacometrics. In:. New York: Academic (SOD),asiatic acid and and glutathione madecassic peroxidase acid derived in from liver Press. 1964. p. 135–65. Centella asiatica had the effect as analgesic and dari eceng gondok Eichornia crassipes 16 Wijayani A, Ummah K, Tjahjani S. Karakterisasi karbosimetil selulosa (CMC) 31. anti-inflammatory. Sancar(Mart) Solms.M, Hantash Indo J T, Chem. Okuyan 2005;5(3):228– B, Apikoglu- misoprostolMoreover, in this preventing study showed gastric thatmucus Centella levels asiaticareduction (400 of mg/kg)aspirin-induced had similar gastric effect ulcer with 9. study is Centella asiatica leaves can prevent Rabus S, Cirakli Z, Gulluoglu MG, et al. treatment of aspirin-induced gastric ulcers. modelgastric inmucus rats (p ≥level 0.05). reduction The conclusion of aspirin- of this Comparative effectiveness of Glycyrrhiza induced gastric ulcer model in rats. glabravs. omeprazole and misoprostol for 20. References 10. GreenAfr J Pharm AP, Lander Pharmacol. JE, Turner 2009;3(12):615– DH. The Effect 1. Valle JD. Peptic Ulcer Disease and Related Disorders. In: Fauci AS, Kasper DL, Longo of Stress and indomethacin, carbenoxolone, JL, et.al., editors. Harrison’s Principles salbutamol, metiamide, zolimidine and of Internal Medicine. 17thed. The US: 11. CF19415 on rat gastric mucus. J Pharm DL, Braunwald E, Hauser SL, Jameson toPharmacol. systems. 1981;33(6):348–52.7th ed. Canada: Brooks/Cole, 1855–72. Sherwood L. Human physiology: from cells 2. McGraw-Hill Companies, Inc.; 2008. p. 12. Shukla A, Rasik AM, Jain GK, Shankar R, Cengage Learning; 2010.

Badan Pengawas Obat dan Makanan asiaticoside isolated from Centella asiatica. Republik Indonesia. Kriteria dan tata Kulshrestha DK, Dhawan BN. In Vitro laksana pendaftaran obat tradisional, obat and In Vivo wound healing activity of 3. herbal terstandar dan fitofarmaka. Jakarta: 13. MacKay D, Miller AL. Nutritional support BPOM; 2005. J Ethnopharmacol. 1999;65(1):1–11. Kementrian Kesehatan Republik Indonesia. Profil kesehatan tahun 2008. 14. Pittellafor wound F, Dutra healing. RC, Junior Altern DD, Lopes Med MTP, Rev. 4. Jakarta: Departemen Kesehatan Republik 2003;8(4):359–77. Indonesia; 2009. Winarto WP, Surbakti M. Khasiat dan Barbosa NR. Antioxidant and cytotoxic manfaat2003. pegagan tanaman penambah daya 15. activities of Centella asiatica (L) Urb. Int J 5. Badaningat. Jakarta:Tenaga Nuklir PT AgroMedia Nasional. Pedoman Pustaka; Mol Sci. 2009;10(9):3713–21. Huangactivities S-S, and Chiu the C-S, mechanisms Chen H-J, Hou of anti- W-C, Sheu M-J, Lin Y-C, et al. Antinociceptive Evid Based Complement Alternat Med. 6. Etik Penggunaan dan Pemeliharaan Hewan HassanPercobaan. MIA. Jakarta: Anti-ulcer BATAN; activity 2011. of Centella inflammation of asiatic acid in mice. 16. Abdulla MA, Al-Bayaty FH, Younis LT, induced Gastric Mucosal Injury in Rats. J Samsuddin2011:2011;895857. L, Somchit N, Israf DA, et al. Somchit MN, Sulaiman MR, Zuraini A, asiatica Leaf Extract Against Ethanol- Effects of Centella asiatica. Indian J 7. Med Plant Res. 2010;4(13):1253–59. Antinociceptive and Antiinflammatory Paget GE, Barnes JM. Toxicity tests. Pharmacol. 2004;36(6):377–80.

Althea Medical Journal. 2015;2(1)