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- In vivo Evaluation of the Pharmacological Activities of Syzygium samarangense (Blume) Merr. L.M. Perry
- Key words
- MATERIALS AND METHODS
- Anti-inflammatory Activity
- CNS Depression Activity
- RESULTS
- Anti-Diarrheal Activity
- Effect on Gastrointestinal Motility
- DISCUSSION
| Advances in Biological Research 8 (3): 107-115, 2014 ISSN 1992-006 © IDOSI Publications, 2014 DOI: 10.5829/idosi.abr.2014.8.3.81225 Corresponding Author: M.D. Badrul Alam, Department of Food Science and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea. Tel: +8801717057701. 107
Graduate School of Biotechnology, Yeungnam University, Geyoungsan, South Korea 1 Department of Pharmacy, Atish Dipankar University of Science and Technology, Dhaka, Bangladesh 2 Department of Clinical Pharmacy and Pharmacology, University of Dhaka, Dhaka, Bangladesh 3 Graduate school of Food Science and Biotechnology, Kyungpook National University, Daegu, South Korea 4 Abstract: Syzygium samarangense is very popular and usually used in the traditional system of medicine. The present studies were carried out to investigate the analgesic, anti-inflammatory, CNS depression as well as the anti-diarrheal activity of the methanolic extract of Syzygium samarangense leaves (MSSL) and barks (MSSB). MSSL and MSSB were used to investigate the analgesic effect by acetic acid induced writhing and formalin induced licking method whereas carrageenan induced inflammation was used for anti-inflammatory activity. CNS depression activities were evaluated in hole-cross and open field test methods. Furthermore castor oil-induced diarrhea and charcoal-induced gastrointestinal motility were used to investigate the anti-diarrheal activity of MSSL and MSSB. MSSBat doses of 100, 200 and 300 mg/kg body weight p.o., significantly (p<0.05) reduced the writhing caused by acetic acid and the number of licks induced by formalin in a dose dependent manner. At 400 mg/kg doses of MSSB showed highest anti-inflammatory activity (% inhibition 72.82%) after 4 hrs. A statically significant CNS depression activity was also observed in both hole cross and open field tests in a dose dependent manner. MSSL and MSSB significantly reduced the frequency and severity of diarrhea in test animals throughout the study period in a dose dependent manner and also showed a significant (p<0.05) reduction in the gastrointestinal motility in charcoal meal test. Altogether, these results suggest that the Syzygium samarangense have good pharmacological effects and provide as a part of scientific support for the use of this species in traditional medicine. Key words: Anti-Inflammatory Analgesic Anti-Diarrheal CNS Syzygium samarangense INTRODUCTION methoxychalcone; six quercetin glycosides: reynoutrin; Syzygium samarangense (syn. Eugenia javanica) one flavanone: (S)-pinocembrin and two phenolic acids: is a plant species in the Myrtaceae, native to an area gallic acid and ellagic acid were isolated from the pulp and that includes the Malay Peninsula and the Andaman seeds of the fruits of S. samarangense [8]. Furthermore, and Nicobar Islands, but introduced in prehistoric times mearnsitrin; 2-C-methyl-5-O-galloylmyricetin-3-O-a- to a wider area [1] and now widely cultivated in the lrhamnopyranoside; desmethoxymatteucinol[9]; 4,6- tropical countries. Leaves of Syzygium samarangense dihydroxy-2-methoxy-3,5-dimethylchalcone;methyl 3-epi- have been reported to have antibacterial [2], antidiabetic betulinate; oleanolic acid; jacoumaric acid; ursolic acid; [3] anti-diarrheal [4], antioxidant [5], analgesic and arjunolicacid[10]; samarangenin A and samarangenin anti-inflammation [6] as well as immunostimulant [7] B[11] were also isolated. In addition from the hexane activities. Three C-methylated chalcones; 2',4'-dihydroxy- extract of the leaves a mixture of a-carotene and ß- 3',5'-dimethyl-6'-methoxychalcone; 2',4'-dihydroxy-3'- carotene;lupeol; botulin;epi-betulinic acid; 2,4-dihydroxy- methyl-6'-methoxychalcone; and
2',4'-dihydroxy-6'- 6-methoxy-3-methylchalcone; 2-hydroxy-4,6-dimethoxy-3- hyperin; myricitrin; quercitrin; quercetin and guaijaverin,
Advan. Biol. Res., 8 (3): 107-115, 2014 108
methylchalcone; 2,4-dihydroxy-6-methoxy- Institutional Animal Ethics Committee, Atish Dipankar 3,5dimethylchalcone; 2,4-dihydroxy-6-methoxy- University of Science & Technology, Dhaka, Bangladesh. 3–methyldihydrochalcone and7-hydroxy-5-methoxy-6,8- Animal treatment and maintenance for acute toxicity and dimethylflavanone; sitosterol and sitosteryl analgesic effects were conducted in accordance with the glucosidewere also isolated [12]. Principle of Laboratory Animal Care (NIH publication Literature reviews indicated that no combined studies No. 85-23, revised 1985) and the Animal Care and Use in analgesic, anti-inflammatory, anti-diarrheal and CNS Guidelines of Atish Dipankar University of Science and depression effects of the bark of Syzygium samarangense Technology, Dhaka, Bangladesh. have so far been undertaken. Taking this in view and as a part of our ongoing research [13] on Bangladeshi
medicinal plants, the present study was aimed to performed in healthy adult male and non-pregnant adult evaluate the comparison of analgesic, anti-inflammatory, female albino Swiss mice (28-32g) divided into different anti-diarrheal and CNS depression activities of the groups. The test was performed using increasing oral methanolic extract of the leaves and bark of Syzygium dose of the MSSL and MSSB (50, 100, 200, 500, 1000
mg/kg body weight p.o.), in 20 ml/kg volume to different MATERIALS AND METHODS allowed to feed ad libitum, kept under regular observation Plant Materials and Extraction: The fresh leaves and bark of Syzygium samarangense were collected from Analgesic Activity Ramna, Dhaka, Bangladesh in July, 2012 and identified by Acetic Acid-Induced Writhing Test: The analgesic DR. M.A. Razzaque Shah PhD, Tissue Culture Specialist, activity of the samples was studied using acetic BRAC Plant Biotechnology Laboratory, Bangladesh and acid-induced writhing model in mice. The animals were the voucher specimen no. maintained in our laboratory divided into eight groups with five mice in each group. for future reference. The both plant materials were Group I animals received vehicle (1% Tween 80 in water, shade-dried with occasional shifting and then powdered p.o.), animals of Group II received Diclofenac-Na at 10 with a mechanical grinder, passing through sieve#40 and mg/kg body weight while animals of groups III, IV, V, VI, stored in an air-tight container. The dried powder material VII and VIII were treated with 100, 200 and 300 mg/kg of leaves (1.0 kg) and barks (1.0 kg) was refluxed with body weight (p.o.) of the MSSL and MSSB, respectively. MeOH for three hours. The total filtrate was concentrated Test samples and vehicle were administered orally 30 min to dryness; in vacuo at 40°C to render the MeOH extract before intra-peritoneal administration of 0.7% v/v acetic 160 g and 180 g for leaves and barks, respectively. acid but Diclofenac-Na was administered intra-peritonially
5 min, the mice were observed for specific contraction of carrageenan were purchased from E. Merck (Germany). body referred to as ‘writhing’ for the next 10 min [14]. Atropine, Loperamide, Diazepam, Indomethacin, Diclofenac-Na were collected from Square
Pharmaceuticals Ltd., Bangladesh. All other chemicals and was determined using the formalin test described by reagents were of analytical grade. Dubuission and Dennis[15]. Control group received 2.5%
dorsal surface of the right hind paw 30 min after either sex weighing about 140-160 g and Swiss Albino administration of MSSL and MSSB (100, 200 and 300 mice (25-30g) were used for assessing biological activity. mg/kg, body weight p.o. respectively) and 15 min after The animals were maintained under standard laboratory administration of Diclofenac Na (10 mg/kg, body weight conditions and had free access to food and water ad p.o.). The mice were observed for 30 min after the injection libitum. The animals were allowed to acclimatize to the of formalin and the amount of time spent licking the environment for 7 days prior to experimental session. injected hind paw was recorded. The first 5 min post The animals were divided into different groups, each formalin injection is referred to as the early phase and the consisting of five animals which were fasted overnight period between 15 and 30 min as the late phase. The total prior to the experiments. Experiments on animals were time spent licking or biting the injured paw (pain behavior) performed in accordance with guidelines of the was
measured with
a stop
watch. test groups. Normal group received water. The mice were for 48 hrs, for any mortality or behavioral changes [13]. 15 min before injection of acetic acid. After an interval of formalin, 20 µl of 2.5% formalin was injected into the
Advan. Biol. Res., 8 (3): 107-115, 2014 109
Anti-inflammatory Activity performed according to the method described by Carrageenan Induced Rat Paw Edema: Long-Evan rats Shoba and Thomas [19]. Briefly, mice fasted for 24 hrs (140-160 g) of both sexes were divided into six groups of were randomly allocated to four groups of five animals five animals each. The test groups received 100, 200 and each. The animals were all screened initially by giving 300 mg/kg body weight p.o. of the extract MSSL and 0.5 ml of castor oil. Only those showing diarrhea were MSSB. The reference group received Indomethacin selected for the final experiment. Group I received 1% (10 mg/kg body weight, p.o.) while the control group CMC (10 ml/kg, p.o.),Groups III, IV, V,VI, VII and VIII were received 3 ml/kg body weight normal saline. After 30 min, treated with 100, 200 and 300 mg/kg body weight (p.o.) 0.1 ml 1% carrageenan suspension in normal saline was of the MSSL and MSSB, respectively. Group II was given injected into the sub-planatar tissue of the right hind paw. Loperamide (3 mg/ kg, p.o.) in suspension. After 60 min, The paw volume was measured at 1, 2, 3 and 4 hr after each animal was given 0.5 ml of castor oil, each animal was carrageenan injection using a micrometer screw gauge. placed in an individual cage, the floor of which was lined The percentage inhibition of the inflammation was with blotting paper which was changed every hour, calculated from the formula: observed for 4 hrs and the characteristic diarrheal % inhibition = (1-D D ) x 100 t/ o where, D was the average inflammation (hind paw edema) into four groups of five mice each and each animal was o of the control group of mice at a given time, D was the given orally 1 ml of charcoal meal (5% activated charcoal t average inflammation of the drug treated (i.e., extract or suspended in 1% CMC) 60 min after an oral dose of drugs reference Indomethacin) mice at the same time [16]. or vehicle. Group I was administered 1% CMC (10 ml/kg
and VIII were treated with 100, 200 and 300 mg/kg body Hole Cross Test: The method used was done as weight (p.o.) of the MSSL and MSSB, respectively. Group described by Takagi et al. [17]. The animals were divided II received atropine sulfate (0.1 mg/kg body weight p.o.) into different groups and each group contains 6 animals. as the standard drug. After 30 min, animals were killed by The control group received vehicle (1% Tween 80 in water light ether anaesthesia and the intestine was removed at the dose of 10 ml/kg p.o.) whereas the test group without stretching and placed lengthwise on moist filter received MSSL and MSSB extract (at the doses of 100, 200 paper. The intestinal transit was calculated as a and 300 mg/kg p.o.) and standard group received percentage of the distance travelled by the charcoal meal Diazepam at the dose of 1mg/kg body weight p.o. orally. compared to the length of the small intestine [20]. Each animal was then placed on one side of the chamber and the number of passages of each animal through the Statistical Analysis: All values were expressed as the hole from one chamber to the other was recorded for 3 min mean ± SEM (Standard Error Mean) of three replicate on 0, 30, 60, 90 and 120 min during the study period. experiments. The analysis was performed by using SPSS
Inc, Chicago). Results related to the reducing power described by Gupta et al. [18]. The animals were divided activities were statistically analyzed by applying the into control standard and test groups (n = 6 per group). Student t-test and p<0.05 were considered to be The control group received vehicle (1% Tween 80 in water statistically significant. All in vivo data are subjected to at the dose of 10 ml/kg p.o.). The test group received the ANOVA followed by Dunnett’s test and p<0.05 were crude extract (at the doses of 100, 200 and 300 mg/kg p.o) considered to be statistically significant. and standard group received Diazepam at the dose of 1mg/kg body weight orally. The animals were placed on RESULTS the floor of an open field (100 cm×100 cm×40 cm hr) divided into a series of squares. The number of squares
visited by each animal was counted for 3 min on 0, 30, 60, aim at establishing the therapeutic index, i.e., the ratio 90, 120, 180 and 240 min during the study period. between the pharmacologically effective dose and the
MSSL and MSSB were safe up to a dose of 1000 mg/kg Castor Oil-Induced Diarrhea: The experiment was (p.o.) body weight. Behavior of the animals was closely droppings were recorded.
body weight p. o.) and animals in groups III, IV, V,VI, VII statistical package for WINDOWS (version 16.0; SPSS lethal dose on the same strain and species. Both extract of Advan. Biol. Res., 8 (3): 107-115, 2014 110
Table 1: Effects of methanolic extract of MSSL and MSSB on acetic acid-induced writhing in mice Groups
Dose (mg/kg) No. of writhing % inhibition Group I
Vehicle 32.77 ± 1.10 Group II 10 8.70 ± 1.30 73.45 a Group III 100 27.09 ± 1.16 17.33 Group IV
200 16.19 ± 2.16 50.59 a
300 11.37 ± 1.96 65.30 a
100 25.02 ± 1.06 23.64 Group VII 200 14.10 ± 1.24 56.97 a
300 9.10 ± 1.04 72.23 a
a (1% Tween 80 in water), group II received Diclofenac Na as standard 10 mg/kg body weight (p.o.), groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight (p.o.) of MSSL and MSSB, respectively. Table 2: Effects of methanolic extract of MSSL and MSSB in the hind paw licking in the formalin test in mice Groups Dose (mg/kg) Early phase (Sec) % protection Late phase (Sec) % protection Group-I Vehicle
37.20 ±1.89 - 40.16 ± 1.82 - Group-II
10 10.82 ±1.13* 70.91 11.03 ± 0.36* 72.53 Group-III 100 30.50±1.19 18.01 31.16 ± 1.82 22.41 Group-IV
200 21.10 ±1.61* 43.27 19.23 ± 1.12* 52.11 Group V
300 16.70 ±1.79* 55.10 15.83 ± 1.22* 60.58 Group VI
100 26.20 ± 1.39 29.56 27.16 ± 1.42 32.37 Group VII 200 15.0 ±1.12* 59.67 12.16 ± 1.99* 69.72 Group VIII 300 11.12 ±1.03* 70.10 11.13 ± 1.36* 72.28 Values are mean ± SEM, (n = 5); * p <0.05 as compared to vehicle control (One way ANOVA followed by Dunnet test). Group I animals received vehicle (1% Tween 80 in water), group II received Diclofenac Na as standard 10 mg/kg body weight p.o., groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight (p.o.) of MSSL and MSSB,respectively. Table 3: Effect of methanolic extract of the MSSL and MSSB on hole cross test in mice Number of Movements ------------------------------------------------------------------------------------------------------------------------------ Group
Dose 0 min
30 min 60 min
90 min Group-I
10ml/kg, 14.60± 1.35 17.00 ± 1.71 17.20 ± 0.96 18.60 ± 1.12 Group-II
1mg/kg, 15.00 ± 1.15 5.20± 0.61* 3.40± 0.57* 1.60± 0.44* Group-III 100 mg/kg 14.80 ± 1.50 13.10 ± 1.11 10.20 ± 0.96 9.60 ± 1.12* Group-IV
200 mg/kg 15.10 ± 1.50 9.40±1.57* 6.20±0.65* 4.80± 1.41* Group-V
300 mg/kg 13.20 ± 0.65 7.80±1.65* 4.60±0.57* 3.10± 2.02* Group-VI
100 mg/kg 13.10 ± 1.50 11.10±1.47 8.20±0.65* 5.10± 1.41* Group-VII 200 mg/kg 14.00±0.61 7.60± 1.14* 5.90±0.57* 4.16± 2.02* Group-VIII 300 mg/kg 14.00± 0.23 5.80± 1.54* 3.79± 0.14* 2.10±0.65* Values are mean ± SEM, (n = 6); * p <0.05, as compared to vehicle control (One way ANOVA followed by Dunnet test). Group I animals received vehicle (1% Tween 80 in water), group II received Diazepam 1 mg/kg body weight, groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight p.o. of the MSSL and MSSB, respectively. Table 4: Effects of methanolic extract of the MSSL and MSSB on open field test in mice Number of Movements ------------------------------------------------------------------------------------------------------------------------------ Group
Dose 0 min
30 min 60 min
90 min Group-I
10ml/kg, 118.4 ± 1.20 118.01±1.30 115.41±0.50 117.48± 1.16 Group-II
1mg/kg, 117.2 ± 1.15 64.65±0.43* 40.85± 0.58* 12.86± 0.86* Group-III 100 mg/kg 110.4 ± 0.81 90.87±1.02* 77.89±1.35* 52.88± 0.02* Group-IV
200 mg/kg 117.8 ± 1.43 75.21± 0.16* 51.61± 0.92* 34.61±0.92* Group-V
300 mg/kg 116.2 ± 1.15 57.25± 0.06* 48.62± 1.12* 31.65±2.32* Group-VI
100 mg/kg 121.2 ± 1.15 81.21± 1.06* 57.11± 1.12* 40.11±1.92* Group-VII 200 mg/kg 111.4 ± 1.81 71.52±1.12* 46.88±1.15* 28.25± 0.62* Group-VIII 300 mg/kg 117.8 ± 1.13 65.91± 0.66* 41.12± 0.42* 13.12±0.22* Values are mean ± SEM, (n = 6); * p <0.05, as compared to vehicle control (One way ANOVA followed by Dunnet test). Group I animals received vehicle (1% Tween 80 in water), group II received Diazepam 1 mg/kg body weight, groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight p.o.of the MSSL and MSSB, respectively. Advan. Biol. Res., 8 (3): 107-115, 2014 111
Table 5: Effect of Methanolic extracts of the MSSL and MSSB extracts on castor oil-induced diarrhea in mice. Treatment Dose Onset of diarrhea (min) Animals with diarrhea No. of faeces in 4 hrs % inhibition of defaecation Group I
10ml/kg, 25.45 ± 1.19 5/5 21.16 ± 0.68 Group II 1mg/kg,
160 ± 0.13 1/5
3.91 ± 0.58 81.52
* ** Group III 100 mg/kg 30.67 ± 2.73 4/5 20.02 ± 1.05 5.38 Group IV
200 mg/kg 51.23 ± 3.03 3/5 13.61± 0.29 35.68 * * Group V 300 mg/kg 65.13 ± 3.03 2/5
8.61± 0.29 59.31
* * Group VI 100 mg/kg 35.83 ± 3.03 3/5 14.81± 0.29 30.00 * Group VII 200 mg/kg 58.61± 2.73 2/5 10.22±1.05 51.70 * ** Group VIII 300 mg/kg 85.23± 3.03 1/5
4.11±0.29 80.57
* ** Values are presented as mean ± SEM, (n=5); p<0.05, respectively, compared to control by student’s t-test. Group I animals received vehicle (1% CMC in * water), group II received Loperamide 10 mg/kg body weight p.o., groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight p. o. of the MSSL and MSSB, respectively. Fig. 1: Effects of the extracts of MSSL and MSSB on carrageenan induced paw edema test. Values are mean ±SEM, (n = 5); *P<0.05 **P<0.005 as compared to vehicle control (One way ANOVA followed by Dunnet test). Group I animals received vehicle (1% CMC in water), group II received Indomethacin 10 mg/kg body weight p.o., groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight (p.o.) of MSSL and MSSB, respectively Fig. 2: Effect on gastrointestinal motility of MSSL and MSSB values are presented as mean ± SEM, (n=5); p<0.05, * respectively, compared to control by student’s t-test. Group I animals received vehicle (1% CMC in water), group II received Atropine sulfate 0.1 mg/kg body weight (p.o.) groups III, IV, V, VI, VII and VIII were treated with 100, 200 and 300 mg/kg body weight (p.o.) of MSSL and MSSB, respectively Advan. Biol. Res., 8 (3): 107-115, 2014 112
observed for the first 3 hr then at an interval of every 4 hrs dependent manner when compared with the untreated during the next 48 hrs. The extract did not cause mortality controls. At 300 mg/kg doses, MSSL showed 59.31% and in mice during 48 hrs observation but little behavioral MSSB 80.57% reduction in the number of fecal episodes, changes, locomotor ataxia, diarrhea and weight loss were whereas Loperamide offered 81.52% protection (Table 5). observed. Food and water intake had no significant difference among the group studied. Effect on Gastrointestinal Motility: With the Analgesic Activity showed significant difference compared with control Acetic Acid Induced Writhing Method: Table 1. Shows (p< 0.05). The intestinal transit of charcoal meal was 75. the effect of both extracts of MSSL and MSSB on acetic 37% in control group, but at 300 mg/kg b.wt. the dose of acid induced writhing in mice. The analgesic activity of MSSB was 27.68% (Fig. 2). MSSL and MSSB was significantly (p<0.05) inhibited writhing response induced by acetic acid in a dose DISCUSSION dependent manner. Formaline Induced Writhing Method: Both MSSL and procedure to evaluate peripherally acting analgesics and MSSB significantly (P<0.05) suppressed the licking represents pain sensation by triggering localized activity in either phase of the formalin-induced pain in inflammatory response mediated by peritoneal mast cells, mice in a dose dependant manner (Table 2). MSSB at the acid sensing ion channels and the prostaglandin dose of 300 mg/kg body weight (p.o.) showed the almost pathways [14]. It is well known that non-steroidal similar analgesic activity against both phases of formalin- anti-inflammatory and analgesic drugs mitigate the induced pain than that of the standard drug diclofenac inflammatory pain by inhibiting the formation of pain Na. mediators at the peripheral target sites where Anti-Inflammatory Activity: Figure 1 represents the significant role in the pain process [21]. MSSL and MSSB anti-inflammatory activity of MSSL and MSSB. Both might have exerted its peripheral antinociceptive action by extracts showed dose dependent anti-inflammatory interfering with the local reaction caused by the irritant or activity and statistically significant (P<0.05). At 300 mg/kg by inhibiting the synthesis, release and/or antagonizing dose, MSSB showed remarkable anti-inflammatory effects the action of pain mediators at the target sites. However, (% inhibition 72.82%) compared with the Indomethacin the biphasic formalin model is represented by neurogenic (% of inhibition 75.02%). (0-5 min) and inflammatory pain (15-30 min), respectively CNS Depression Activity pains by the extract might imply that it contains active Hole-Cross Test: In the Hole- cross test, MSSL and analgesic principle that may be acting both centrally and MSSB extracts exhibited statistically significant (P<0.05) peripherally. This is an indication that the extract can be of decrease in the movements of the test animals at all used to manage acute as well as chronic pain. Since dose levels tested and followed a dose-dependent flavonoids inhibiting the writhing will have analgesic response. The depressing effect was most intense during effect preferably by inhibition of prostaglandin synthesis, the second (60 min) and third (90 min) observation periods a peripheral mechanism of pain inhibition [23] and in both extract (Table 3). previous studies [6, 9] have evidenced the presence of Open Field Test: Results of the hole-cross test followed augmenting the analgesic activity. a similar trend to the ones observed in the open-field test. Carrageenan induced edema has been commonly They were statistically significant for all dose levels and used as an experimental animal model for acute followed a dose-dependent response. The depressing inflammation and is believed to be biphasic. The early effect was most intense during the second (60 min) and phase (1-2hrs) of the carrageenan model is mainly third (90 min) observation periods (Table 4). mediated by histamine, serotonin and increased synthesis Anti-Diarrheal Activity The late phase is sustained by prostaglandin release and Castor Oil-Induced Diarrhea: The extracts significantly mediated by bradykinin, leukotrienes, polymorphonuclear reduced the number of diarrheal episodes in a dose cells and prostaglandins produced by tissue macrophages gastrointestinal transit experiment, the treated groups Acetic acid induced writhing response is a sensitive prostaglandins and bradykinin are proposed to play a [22]. The suppression of neurogenic and inflammatory flavonoid in S. samarangensemay play the vital role for of prostaglandins in the damaged tissue surroundings. Advan. Biol. Res., 8 (3): 107-115, 2014 113
[13]. Since the extract significantly inhibited paw edema which was used as a diarrhea inducing agent in the induced by carrageenan in the second phase and this experimental protocol. Several mechanisms have been finding suggests a possible inhibition of cyclooxygenase previously proposed to explain the diarrheal effect of synthesis by the extracts and this effect is similar to that castor oil including inhibition of intestinal Na , K - produced by non-steroidal anti-inflammatory drugs ATPase activity to reduce normal fluid absorption, such as Indomethacin, whose mechanism of action is activation of adenylate cyclase or mucosal cAMP inhibition of the cyclooxygenase enzyme. Flavonoids, mediated active secretion, stimulation of prostaglandin Cycloartenyl stearate, lupenyl stearate, sitosteryl stearate formation, platelet activating factor and recently nitric and 24-methylenecycloartanyl stearate from the oxide has been claimed to contribute to the diarrhoeal Syzygiumsamarangense exhibited potent analgesic and effect of castor oil [27]. However, it is well evident that anti-inflammatory activities at different doses form [6] castor oil produces diarrhea due to its most active and also well known for their ability to inhibit pain component recinoleic acid which causes irritation and perception as well as anti-inflammatory properties due to inflammation of the intestinal mucosa, leading to release their inhibitory effects on enzymes involved in the of prostaglandins, which results in stimulation of production of the chemical mediator of inflammation. secretion [28]. Previously flavonoids isolated from This hypothesis is strongly supported by the previous
study, which has shown that MSSL and MSSB possess spasmolytic activity [29]. These indicate that the presence anti-inflammatory activity due to the presence of high of compounds with spasmolytic and calcium antagonist flavonoids content [24]. activity may be responsible for the anti-diarrheal effect. In Locomotor activity considered as an increase in the small intestinal transit test, both extracts of MSSL and alertness and decrease in locomotor activity indicated MSSB suppressed the propulsion of charcoal marker in a sedative effect [25]. Extracts of MSSL and MSSB dose dependent manner. This finding suggests that the decreased locomotor activity indicates its CNS depressant extracts act on all parts of the intestine. A decrease in the activity. Gamma-aminobutyric acid (GABA) is the major motility of gut muscles increases the stay of substances inhibitory neurotransmitter in the central nervous system. in the intestineallows better water absorption [30]. It is Different anxiolytic, muscle relaxant, sedative-hypnotic therefore presumed that the reduction in the intestinal drugs are elucidation their action through GABA, propulsive movement in the charcoal meal model may be therefore it is possible that extracts of MSSL and MSSB due to antispasmodic properties of the extracts. may act by potentiating GABAergic inhibition in the CNS via membrane hyper-polarization which lead to a decrease
in the firing rate of critical neurons in the brain or may be due to direct activation of GABA receptor by the extracts. In summary, the methanolic extract of Many research showed that plant containing flavonoids,
saponins and tannins are useful in many CNS disorders anti-inflammatory, CNS depression and anti-diarrheal [26]. Earlier investigation on phyto-constituents and properties. Further investigations are required to find plants suggests that many flavonoids and neuroactive the active component of the extract in order to confirm steroids were found to be ligands for the GABA the mechanism of action in the development of a A receptors in the central nervous system; which led to potent analgesic, anti-inflammatory CNS depression and assume that they can act as Benzodiazepine like molecules anti-diarrheal reagent. [25].
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