{"id":50785,"date":"2023-09-30T10:16:40","date_gmt":"2023-09-30T10:16:40","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=50785"},"modified":"2023-10-07T11:54:01","modified_gmt":"2023-10-07T11:54:01","slug":"evaluation-of-the-analgesic-activity-of-ethanolic-extract-of-manilkara-zapota-seeds-in-experimental-animal-model","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no3\/evaluation-of-the-analgesic-activity-of-ethanolic-extract-of-manilkara-zapota-seeds-in-experimental-animal-model\/","title":{"rendered":"Evaluation of the Analgesic Activity of Ethanolic Extract of Manilkara Zapota Seeds in Experimental Animal Model"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

The International Association for the Study of\nPain (IASP) has defined pain as “an unpleasant sensory and emotional\nexperience associated with actual or potential tissue damage”.1<\/sup>\nPain could be more well-defined but almost associated with all medical\nconditions. It is often provoked by noxious stimuli, which can be of external\nor internal origin.2<\/sup> Pain obstructs daily activities as well as the\ngeneral functioning of the body. The majority of the patients consult the\nphysician because of the pain. It is the foremost & chief symptom of many\ndiseases.<\/p>\n\n\n\n

Central and peripheral analgesics relieve pain\nwithout affecting consciousness. Analgesics are divided into opioids and\nNon-steroidal anti-inflammatory Drugs (NSAID). Opioids are indicated in\ndeep-seated visceral pain, while NSAIDs are in pain associated with inflammation\n& tissue injury.3<\/sup> NSAID and opioids use remain the mainstay for\nthe treatment of pain. However, both groups of drugs are well known for their\ncommon & severe side effects. The studies conclude that opioids lead to\ntolerance, physical dependence, and addiction, whereas NSAIDs lead to gastrointestinal\nbleeding & ulcers.4<\/sup><\/p>\n\n\n\n

For that, looking for an alternative compound\nwith minimal or no side effects for pain management is necessary. Medicinal\nplants could be the better option to overcome these side effects.<\/p>\n\n\n\n

Conventional drug therapy for diseases can\noccasionally have severe adverse effects. All over the world, there is a trend\nto explore traditional medicines. Siddha is an old medicine branch famous\namongst the Tamil-speaking people. There are various sources of the drugs. These\ncan be broadly classified into plants, animals, microorganisms & minerals.\nThere is a need to identify the accurate source of the drug, which can give\ngood results with minimal side effects. The availability of plant sources is\nmore as compared to the other sources. The newly discovered active compound\nshould have higher efficacy, safety, and fewer side effects. <\/p>\n\n\n\n

Medicinal plants contain many chemical substances\nwhich have possible and multiple therapeutic uses. Medical medicinal plants\nhave been used for therapeutic use in many disease conditions worldwide. Still,\nthe active compounds from the medicinal plants are used to prepare drugs. The\nprincipal advantage of herbal medicine is that they are safer than synthetic\nmedicine as they have fewer side effects and are cost-effective, which would be\naccepted by developing countries like India.<\/p>\n\n\n\n

From ancient times, different parts of medicinal\nplants were in use to cure different diseases, especially in India.5<\/sup>\nManilkara zapota belongs to the Sapotaceae family, which is commonly called by\ndifferent names in different languages, such as Sapota in Hindi, American bully\nin English, Simaiyiluppai in Tamil, and Sapotasima in Telugu.<\/p>\n\n\n\n

Manilkara zapota is a big tree with rough, lumpy\nshady grey bark and a thick crown. Leaves are elliptic-oblong, 7.5-12.5 cm\nlong, shining from both sides with small, discrete secondary branching nerves.\nFlowers are lengthy. Fruits are round with 5-6 big seeds and light brown pulp.\nThe seeds are hard and brown or black with one white margin. The seeds contain\nphytochemicals like alkaloids, flavonoids, saponins, tannins, steroids,\nglycosides, and phenolic compounds. Hydrocyanic acid is also present in seeds,\nso it should be removed before eating the fruit.6 <\/sup>  <\/p>\n\n\n\n

Seeds of Manilkara zapota have been evidenced to\nhave diuretic,7<\/sup> antibacterial,8<\/sup> hypoglycemic,9<\/sup>\nand anthelmintic activity.10<\/sup> Bark extract has been assessed for\nantimicrobial and anticancer activities.11<\/sup> The plant leaves hold\nanalgesic and anti-inflammatory,12 <\/sup>antioxidant, hypoglycemic, and hypocholesterolemic\nactivities.13 <\/sup>The roots of the plant are found to have hypoglycemic\nactivity.14<\/sup><\/p>\n\n\n\n

Although many pharmacological investigations have\nbeen carried out based on the constituents present in it, a lot more can still\nbe explored and utilized in a therapeutic manner.<\/p>\n\n\n\n

The anti-inflammatory activity of seeds of\nManilkara zapota has already been demonstrated in experimental animals.15<\/sup>\nAs pain always accompanies inflammation, we have set forward a study to\ndiscover the analgesic activity of the ethanolic extract of Manilkara Zapota in\nanimal models using Eddy’s hot plate method.<\/p>\n\n\n\n

Aims and Objectives<\/strong><\/p>\n\n\n\n

To\nexplore the analgesic activity of ethanolic extract of Manilkara zapota seeds\nusing Eddy’s hot plate method.<\/p>\n\n\n\n

To\ncompare the effect of ethanolic extract of Manilkara zapota seeds with the\ncommonly used analgesic drug aspirin.<\/p>\n\n\n\n

Material and Methods<\/strong><\/p>\n\n\n\n

This\nstudy was conducted after the approval of the Institutional Ethics Committee\n(IEC) (IAEC\/SSSMCRI\/2018-05). The study followed the Helsinki Declaration. The\nseeds of Manilkara zapota were obtained from a forest near the experimental\ninstitute. These fresh seeds were thoroughly cleaned with water to remove dirt.\nSeeds were dried for one week in the shed. After complete drying, the seeds\nwere powdered. The powder was packed in a sealed bottle and stored in a dry,\ncool, and dark place.<\/p>\n\n\n\n

The\nextract is prepared with the help of Soxhlet’s apparatus (hot extraction) and\nextracted with 95% ethanol at a temperature of 60\u00b0-80\u00b0C. Ethanolic extract was\nsubjected to filtration. It was then dried under reduced pressure to obtain a\nsolid mass that was free from the solvent. The dried extract was dissolved in\ndistilled water.<\/p>\n\n\n\n

12\nAdult Wistar albino rats of weight ranging from 235 to 255 g were selected for\nthis study. Following their procurement, they were kept at 24\u00b0C temperature and\n55\u201365 % humidity to acclimatize and were provided a pellet meal and water as\nneeded. The animals were divided into the following three groups, each with\nfour animals.<\/p>\n\n\n\n

The\nanimals were grouped into the following three groups of 4 animals in each\ngroup.<\/p>\n\n\n\n

Group\nI: Control (normal saline), <\/p>\n\n\n\n

Group\nII: Standard (Aspirin 25mg\/kg) <\/p>\n\n\n\n

Group\nIII: Ethanolic extract of Manilkara zapota (200mg\/kg)<\/p>\n\n\n\n

Normal\nsaline, Aspirin & Ethanolic extracts of Manilkara zapota were given orally\nto animals. The dose of Aspirin & Ethanolic extract of Manilkara zapota was\ncalculated according to the weight of each animal.<\/p>\n\n\n\n

This\nstudy was carried out by using Eddy’s hot plate method. The individual animal\nwas placed on a plate and maintained at a temperature of 55\u00b0C.16<\/sup>\nFollowing the administration of normal saline, standard medication, and\nextract, the reaction time was recorded at 0 min, 30 min, 60 min, and 90\nminutes. Reaction time was defined as the time animals responded to the pain\nstimulus by jumping or licking their paws. The reaction time was set at 45\nseconds to prevent harm to an animal’s paw.17<\/sup><\/p>\n\n\n\n

Data\nwere entered on a Microsoft Excel spreadsheet and statistically analyzed\nstatically using the SPSS-20 software. Unpaired student t-tests were used to\ncompare reaction times. A p-value of 0.05 or less than 0.05 is indicated as\nstatistically significant.<\/p>\n\n\n\n

Observations and Results<\/strong><\/p>\n\n\n\n

The mean reaction time of all three groups, i.e., control, standard & test groups, at all time points is illustrated in Table 1. Compared to animals treated with saline, the mean reaction time of the aspirin and extract-treated group was increased, as shown in Figure 1.<\/p>\n\n\n\n

Table 1: Comparison of mean reaction time for all the three groups<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

Treatment<\/strong><\/p>\n<\/td>\n

\n

Mean reaction time (seconds)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

0 minutes<\/strong><\/p>\n<\/td>\n

\n

30 minutes<\/strong><\/p>\n<\/td>\n

\n

60 minutes<\/strong><\/p>\n<\/td>\n

\n

90 min<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

28.25<\/p>\n<\/td>\n

\n

27.00<\/p>\n<\/td>\n

\n

25.25<\/p>\n<\/td>\n<\/tr>\n

\n

Standard<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

39.00<\/p>\n<\/td>\n

\n

40.75<\/p>\n<\/td>\n

\n

34.25<\/p>\n<\/td>\n<\/tr>\n

\n

Test<\/p>\n<\/td>\n

\n

30.25<\/p>\n<\/td>\n

\n

36.25<\/p>\n<\/td>\n

\n

38.50<\/p>\n<\/td>\n

\n

38.50<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 1: comparison of mean reaction time for all the three groups<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

The mean reaction time for normal saline-treated animals was found to be decreased throughout 90 minutes. The mean reaction time for aspirin-treated & extract-treated animals was increased throughout 60 min, but after that, it was found to be declined. Compared to animals treated with saline, the reaction time was significantly increased in both group 2 and group 3. The result of this is depicted in Table 2, figure 2, table 3 & figure 3.<\/p>\n\n\n\n

Table 2: Analgesic effect of aspirin by hot plate method<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Treatment<\/strong><\/p>\n<\/td>\n

\n

Mean reaction time (seconds)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

0 minutes<\/strong><\/p>\n<\/td>\n

\n

30 minutes<\/strong><\/p>\n<\/td>\n

\n

60 minutes<\/strong><\/p>\n<\/td>\n

\n

90 min<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

28.25<\/p>\n<\/td>\n

\n

27.00<\/p>\n<\/td>\n

\n

25.25<\/p>\n<\/td>\n<\/tr>\n

\n

Standard<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

39.00 *<\/p>\n<\/td>\n

\n

40.75 *<\/p>\n<\/td>\n

\n

34.25 *<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 2: Analgesic effect of aspirin by hot plate method<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 3: Analgesic effect of Manilkara zapota seeds extract by hot plate method<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Treatment<\/strong><\/p>\n<\/td>\n

\n

Mean reaction time (seconds)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

0 min<\/strong><\/p>\n<\/td>\n

\n

30 min<\/strong><\/p>\n<\/td>\n

\n

60 min<\/strong><\/p>\n<\/td>\n

\n

90 min<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

28.25<\/p>\n<\/td>\n

\n

27.00<\/p>\n<\/td>\n

\n

25.25<\/p>\n<\/td>\n<\/tr>\n

\n

Test<\/p>\n<\/td>\n

\n

30.25<\/p>\n<\/td>\n

\n

36.25 *<\/p>\n<\/td>\n

\n

38.50 *<\/p>\n<\/td>\n

\n

38.50 *<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 3: Analgesic effect of <\/strong>Manilkara zapota seeds extract by hot plate method.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

When the analgesic activity of aspirin and extract was compared, it was found that there was less increase in the reaction time of the group treated with extract group compared to that of the aspirin-treated group. The reaction time of the extract-treated group at 30 min was found to increase significantly compared with the aspirin-treated group. Whereas the mean reaction time of extract treated group at 60 min & 90 min was not found to be significantly increased in comparison with the standard, as shown in Table 4 & Figure 4<\/p>\n\n\n\n

Table 4: Comparison of Analgesic effect of Aspirin &ethanolic extract of Manilkara zapota seeds<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Treatment<\/strong><\/p>\n<\/td>\n

\n

Mean reaction time (seconds)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

0 minutes<\/strong><\/p>\n<\/td>\n

\n

30 minutes<\/strong><\/p>\n<\/td>\n

\n

60 minutes<\/strong><\/p>\n<\/td>\n

\n

90 minutes<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Standard<\/p>\n<\/td>\n

\n

30.50<\/p>\n<\/td>\n

\n

39.00 #<\/p>\n<\/td>\n

\n

40.75<\/p>\n<\/td>\n

\n

34.25<\/p>\n<\/td>\n<\/tr>\n

\n

Test<\/p>\n<\/td>\n

\n

30.25<\/p>\n<\/td>\n

\n

36.25 #<\/p>\n<\/td>\n

\n

38.50<\/p>\n<\/td>\n

\n

38.50<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 4: Comparison of Analgesic effect of Aspirin &ethanolic extract of Manilkara zapota seeds<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

Recently developed allopathic drugs still exhibit mild to severe\nside effects. Various plants are still unique because of having fewer adverse\neffects. Therefore, the analgesic effect of plant products has been studied\nsystematically.<\/p>\n\n\n\n

As pain is the most common manifestation of most of the\ndiseases. Many synthetic NSAIDs have been used as the primary analgesic, with common\nadverse side effects. Therefore, It is beneficial to investigate an alternative\nanalgesic therapy.<\/p>\n\n\n\n

Analgesics relieve pain selectively without affecting it by\nacting on the central or peripheral nervous system. Centrally-acting analgesics\nlower the pain threshold and alter the body’s typical physiological reaction to\npain. Conversely, peripherally acting analgesics prevent impulse generation at\nthe chemoreceptor site for pain.18<\/sup><\/p>\n\n\n\n

Eddy’s hot plate method is the animal model employed for\nscreening analgesic activity in this study. In this experimental model for\nanalgesia, an increase in reaction time is an important measure or parameter of\ncentral and peripheral analgesic activity.19 <\/sup><\/p>\n\n\n\n

As mentioned earlier, many animal experiments have been conducted\nand proven many activities of Manikara Zapota, according to Ayesha Khan et al.\nThe whole plant extract of Manilkara zapota Linn possesses analgesic activity.\nTherefore, in this study, particularly seeds of the Manilkara zapota plant were\nscreened for analgesic effects by using Eddy’s hot plate.<\/p>\n\n\n\n

Manilkara zapota seeds extract showed a moderate to significant\nanalgesic effect, which was proved by the significant increase in the reaction\ntime, as depicted in Table 3 and Figure 3. Compared to the control, the\nextract’s reaction time was similar at 0 min and slowly increased from 30, 60,\nand 90 min. The increase in mean reaction time of the extract was statistically\nsignificant (* p< 0.0001) at 30 min,60 min, and 90 min compared with that of\nthe control. From this, it can be stated that the crude extract of Manilkara\nzapota seeds is an effective analgesic agent.<\/p>\n\n\n\n

When the analgesic activity of aspirin and Manilkara zapota\nextract were compared, it was observed that there was less increase in the\nreaction time of the group treated with extract compared to that of the\naspirin-treated group at 30 min & 60 min points of time. The Significant\ndifference (# p< 0.001) in the analgesic activity of aspirin and the extract\nwas found at 30 min. The analgesic effect of the extract-treated group at 90\nmin was found to be the same, but the reaction time for aspirin at 90 min was\ndecreased compared to the earlier values, as shown in Table 4 & Figure 4.\nFrom this, we can state that the analgesic activity of aspirin has decreased earlier\nthan that of the extract of seeds.<\/p>\n\n\n\n

The observed analgesic effect of Manilkara zapota seed is\nattributed to the crude extract. Any crude extract contains many phytochemicals\nsuch as steroids, alkaloids, flavonoids, phenolic compounds, etc. Furthermore,\nthe analgesic activity of seed extract may be attributed to the specific\nphytochemicals in the extract. A single constituent or more than one\nconstituent may be responsible for the analgesic effect. Preliminary\nphytochemical screening of crude extract is needed. There is a need for\nadditional studies to identify the actual phytochemicals in the crude extract\nof Manilkara zapota seeds, which are responsible for analgesic activity.<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

The extract under study had a significant analgesic effect compared with the analgesic activity of control in an established analgesic screening method. The analgesic effect was peaking at 60 minutes and 90 minutes. Treating 200mg\/kg of ethanolic extract of Manilkara zapota seeds increases the reaction time on Eddy’s hot plate. Hence, the present study concluded a significant analgesic effect of Manilkara zapota extract. Henceforth, the preparation of Manilkara zapota seed extract could be used as one of the drugs in multi-drug therapy for pain relief. Though, more research is needed in separating and describing the active compound or compounds responsible for the analgesic effect. Further work is also needed for the determination of the exact mechanism of action. <\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

Authors would like to acknowledge the ICMR the ICMR as this was STS ICMR project (REF ID \u2013 2018 – 00245)<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

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     CrossRef <\/a><\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

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