Antinociceptive activity of Karpasa Beeja

L Abhiram

doi:http://dx.doi.org/10.21760/jaims.7.11.8

Antinociceptive activity of Karpasa Beeja

Abhiram L.^1*

DOI: http://dx.doi.org/10.21760/jaims.7.11.8

^1* L Abhiram, Post Graduate Scholar, Department of Dravya Guna Vignana, Ramakrishna Ayurvedic Medical College, Yelahanka, Bengaluru, Karnataka, India.

Background: Karpasa Beeja (cotton seeds) is a drug which is been used since Samhita period, it has Vatahara and Vedanasthapana properties. Acharya Charaka includes Karpasa in Brimhaniya Gana while Acharya Susruta includes it in Vatasamsamana Varga. Cotton seeds are a rich source of cattle feed. It is also widely used in preparation of Ayurvedic medicines. The present study is highly relevant in searching experimentally for its Anti-nociceptive activity in cotton seeds and its pharmacognostical, phytochemical, HPLC+TLC profile. Aim and objective: An experimental evaluation of Karpasa beeja for its Antinociception activity. Results: Acetic acid induced writhing test, mice was treated with paracetamol (72.5mg/Kg) and test Karpasa Beeja drug Kashaya (12.48ml/kg,) and distilled water as control group in a dose of 1ml. The efficacy of test substances for analgesic activity was evaluated in the swiss albino mice. The analgesic activity of Karpasa Beeja Kashaya was studied in swiss albino mice by administering the test substance orally. The writhing effect was induced by 1% V/V glacial acetic acid solution in mice pre-treated with test substance. The number of abdominal writhing were measure over 20 min after injection of acetic acid. Finally, percentage inhibition was calculated. Conclusion: In this test substance it is found that the percentage inhibition of pain is 38.89 % hence it can be concluded that the test substance is known to possess analgesic activit.

Keywords: Karpasa beeja, Gossypium hirsutum, Analgesic, Anti-nociceptive, HPLC+TLC

Corresponding Author	How to Cite this Article	To Browse
L Abhiram, Post Graduate Scholar, Department of Dravya Guna Vignana, Ramakrishna Ayurvedic Medical College, Yelahanka, Bengaluru, Karnataka, India. Email:	L Abhiram, Antinociceptive activity of Karpasa Beeja. J Ayu Int Med Sci. 2022;7(11):46-51. Available From https://www.jaims.in/jaims/article/view/2167

Introduction

Vedana is often considered as a synonym of Vyadhi or disease. This may be because of the fact that almost all the diseases are associated with one or another type of pain. The concept of Vedana in Ayurveda, has a theoretical and philosophical background apart from its clinical importance. Pain is commonly referred to as Vedana. But Vedana is a much broader concept than merely pain. Vedana (Samvedanam- Amarakosam) literally means perception. This may be a pleasant one (Sukham) or an unpleasant one (Dukha). Sukha and Dukha are Gunas attributed to Atma. This perception may come from Sarira, Manas or Atma. The unpleasant feeling, whatever may be its seat is called as Roga or Vikara. The objective of Chikitsa or treatment is to overcome this Ruja the Roga. No other symptom may have got as much importance as the symptom pain as it is the most distressing and disturbing of all symptoms. Vedana, when considered in a broader sense, each Dosha produces its own type of Vedana. For e.g., Daha attributed to Pitta; Toda, Ruja are attributed to Vata and Kandu, Supti are attributed to Kapha. But in a clinical point of view, patient refers to pain as Vedana. This Vedana is especially attributed to Vata. Even this Vedana is of different permutations and combinations. Vedana is a purely subjective experience and only the person who experiences it can explain it. Our Acharyas have tried their best to put the different types of pain into objective terms by the use of certain similies like Ankusavat, Vriscikadagdamiva, Soola etc. but the relevance of such similies in the modern era are now questionable because in olden days people were familiar with the pain caused by the bite of Vriscika or that caused by an Ankusa but now people are less accustomed with such injuries. The subjectivity of pain not only lies in its type but also in its severity. Caraka in his Vimanastana explains about Guru Rogi and Laghu Rogi. The Gurutva or Laghutva of Vyadhi depends on the Satva and Bala of the Rogi. Vedana is primarily considered as an attribute of Vata though Pitta and Kapha also manifests their own types of Vedana.

Pain is a submodality of somatic sensation. The word "pain" is used to describe a wide range of unpleasant sensory and emotional experiences associated with actual or potential tissue damage. Nature has made sure that pain is a signal, we cannot ignore.

In pain the information is transmitted to the CNS via three major pathways. Most ailments of the body cause pain. The ability to diagnose different diseases depends to a great extent on the knowledge of the different qualities and causes of pain. Sensitivity and reactivity to noxious stimuli are essential to the wellbeing and survival of an organism. Pain travels through redundant pathways, ensuring to inform the subject: “Get out of this situation immediately.” Without these attributes, the organism has no means to prevent or minimize tissue injury. Individuals congenitally insensitive to pain are easily injured and most of them die at an early age. For thousands of years, physicians have tried to treat pain without knowing the details of the ways in which pain is signalled from the injured part of the body to the brain, or the ways in which any of their remedies worked.

Recent discoveries about how the body detects, transmits, and reacts to painful stimuli, have allowed physicians to relieve both acute and chronic pain.

Antinociception is the action or process of blocking the detection of painful or injurious stimulus sensory neurons. Pain is termed nociceptive (nocer - to injure or to hurt in Latin), and nociceptive means sensitive to noxious stimuli. Noxious stimuli are stimuli that elicit tissue damage and activate nociceptors. Nociceptors are sensory receptors that detect signals from damaged tissue or the threat of damage and indirectly also respond to chemicals released from the damaged tissue. Nociceptors are free (bare) nerve endings found in the skin, muscle, joints, bone and viscera. Recently, it was found that nerve endings contain transient receptor potential (TRP) channels that sense and detect damage. The TRP channels are similar to voltage gated potassium channels or nucleotide gated channels, having 6 transmembrane domains with a pore between domains 5 and 6. They transduce a variety of noxious stimuli into receptor potentials, which in turn initiate action potential in the pain nerve fibers. This action potential is transmitted to the spinal cord and makes a synaptic connection in lamina I and II. The cell bodies of nociceptors are mainly in the dorsal root and trigeminal ganglia. No nociceptors are found inside the CNS. Nociceptors are not uniformly sensitive. They fall into several categories, depending on their responses to mechanical, thermal, and chemical stimulation liberated by the damage, tumor, and inflammation.

So anti-nociception can be called as peripheral analgesics activity. Many of the peripheral analgesics possess anti-inflammatory property and in some cases antipyretic activity. For many of them the mode of action has been elucidated as an inhibition of cyclooxygenase in the prostaglandin pathway. It can be tested by using Writhing test.

The word Karpasa in Sanskrit denotes the cotton plant or the cotton tree. The use of Karpasa as medicine was not seen in the Vedic period. Only a very few descriptions about it was seen in the Samhita period. The identity of the plant mentioned in Samhita is also controversial because synonyms are used and also while checking the context of mentioning it creates a doubt whether the mentioning is about cotton plant. Elaborate descriptions of Karpasa are seen from the Nighantu period onwards. Karpasa Beeja (cotton seeds) is Vatahara and has got Vedana Sthapana property according to Ayurvedic classics, which gave the base for choosing the study drug for these activities. Acharya Caraka includes Karpasa in Brimhaṇiya Gana^[2] while Acharya Susruta includes it Vata Samsamana Varga.^[3] Karpasa Beeja has Snigdha Guṇa, Madhura Rasa, Madhura Vipaka, Kincit Ushna Virya and Vataharakarma.

Materials and Methods

Cotton seeds were authenticated and purchased from Calicut. The Kaṣhaya of Cotton seeds were prepared at Invivo Biosciences Bengaluru 154 Kodigehalli Village, Magadi road Bengaluru-560091 as per the classical reference of Sarangadhara Samhita, Madhyamakhaṇda, Kashaya Kalpana. During experimentation, on everyday basis, Cotton seeds (Gossypium hirsutum Linn.) were weighed separately over accurate digital weighing machine in the quantity of 20 grams each. Then the seeds were pounded using a metal mortar and pestle and made into moderately crushed seeds. Distilled water was taken 16 times that of weighed drug which is 320ml Then, the pounded drugs were transferred separately into two stainless steel cooking bowls, 320ml of water was added to each of the bowls and kept for boiling over low flame reducing to 1/8th to obtain the quantity of 40 ml of Kashaya each.

Total 18 healthy Swiss albino mice weighing between 120-250gms will be taken and divided randomly into three groups, each containing six rats, maintained under a constant 12hr light

and dark cycle at 22-24 and at 45%-55% relative humidity. Animals were feed with Pelleted rodent from VRK nutrition solutions. Deep bore well water passed through charcoal filters and exposed to UV rays and water in polypropylene water bottles were provided to the animals. The Animal Ethical committee has approved for experiment on animals. The experimental protocol was approved on 25/02/2022 by the Institutional Animal Ethical Committee (IAEC).

Table 1: Group Allocation

Group No	Treatment group	Dose	No. of animals	Animal numbers
1	Control	-	6	1	6
2	Positive control Paracetamol suspension)	100mg/kg	6	7	12
3	Kashaya of cotton seeds -Therapeutic Dose	12.48ml/kg	6	13	18

To the group 1, distilled water administered to serve as control. Group 2 was taken as the standard and administered with aqueous suspension standard drug Paracetamol, (100mg/kg body weight).

Group 3 were administered with test drug cotton seed Kashaya, for all test groups Kaṣhaya was administered once daily for 7 consecutive days. On the 7^th day 1 hour after the administration of drug acetic acid in a dose of 1 ml/100g body weight (1%v/v solution) was injected intra peritoneal to each mouse. Each mouse was placed in separate per specs boxes under observation immediately after the acetic acid injection and the number of abdominal constrictions (writhes) were counted over a period of 15 mins for each animal. (Analgesic effect was recorded by counting the number of writhing syndrome after the injection of acetic acid for a period of 15 minutes). For scoring purposes, a writhe was indicated by stretching of the abdomen by simultaneous stretching of at least one hind limb. The latency of onset and number of writhing was noted. The mean number of writhes and latency of onset of writhing by the trial groups, standard group & control group were calculated and compared statistically.

Percentage of inhibition was evaluated using the formula: average writhes in the test group minus writhes in the control group divided by writhes in the control group times 100. The time period with the greatest percent of inhibition was considered as peak time.

The experimental data were expressed as mean ± SEM. The data obtained was analyzed by using one way analysis of variance (ANOVA) followed by POST HOC TUKEY KRAMER TEST for determining the level of significance of the observed effects. A “P value” of less than 0.05 was considered statistically significant.

Graph Pad In Stat-3 and SPSS graph pad was used for statistical analysis of the generated data.

Observations and Results

Table 2: No. of writhing in Acetic acid induced writhing test

Group	Animal No.	Writhing response	% Inhibition
Control	1	32
	2	35
	3	30
	4	28
	5	38
	6	35
Mean		33.00
SEM		1.51
Standard	7	15	56.06
	12	18
Mean		14.50
SEM		1.31
Test	13	20	38.89
	14	23
	15	18
	16	20
	17	17
	18	23
Mean		20.17
SEM		1.01

Graph 1: No. of writhing in acetic acid induced writhing test

Table 3: Descriptives of No. of writhing in each group.

Groups	Mean	Std. error
Control	33	18.26
Standard	14.50	1.31
Karpasabeeja	20.17	1.01

The efficacy of test substances for analgesic activity was evaluated in the Swiss albino mice. The analgesic activity of Karpasa Beeja Kashayam was studied in Swiss albino mice by administering the test substance orally. The writhing effect was induced by 1% V/V glacial acetic acid solution in mice pre-treated with test substance. The number of abdominal writhing were measure over 20 min after injection of acetic acid. Finally, percentage inhibition was calculated.

Discussion

Acetic acid induced writing test was done to find out the central and peripheral analgesic activity of cotton seeds.

Acetic acid causes pain by liberating endogenous substances such as serotonin, histamine, prostaglandins (PGs), bradykinins and substance P, which enhances inflammatory pain by increasing capillary permeability. Local peritoneal receptors are postulated to be involved in the abdominal constrictions response.

The observation was done in two parts that is latency of onset of writhing as well as number of writhing. The latency of onset was considerably increased by the test drug than the standard drug but it was not statistically significant. While the test drugs showed statistically significant decrease in the number of abdominal constrictions and stretching.

This model as mentioned earlier is for predicting analgesic effect with peripheral mode of action, The writhing observed in this model is mediated by formation and release of phlogistic mediators prostaglandin and bradykinin. The observed effect may be due to decrease in the formation of these mediators. It can also be due to up regulation of the receptor activity of these mediators. Pain, being the most unpleasant sensory and emotional experience, needs utmost attention and treatment. In this study Karpasa Beeja Kashaya^[5] was used to treat the pain induced by chemical agent named acetic acid. Acetic acid induced writhing

in mice attributed visceral pain elicited by triggering localized inflammatory response resulting from release of free arachidonic acid from tissue phospholipid via cyclooxygenase (COX), and prostaglandin biosynthesis. In this study it was found that Karpasa Beeja Kashaya at the dose of 12.48mL/kg body weight showed analgesic activity. The % inhibition of pain is 38.8% in Karpasa Beeja Kashayam when compared against control. The analgesic activity of the test substance may be due to the presence of analgesic principles acting through prostaglandin pathways or otherwise. Further fractionation of the test substance in different solvent may help in understanding the mechanism of analgesic activity.

Order of anti-nociceptive activity of test sample Paracetamol > Karpasa Beeja Kashaya

Writhing behaviour highly significant results were obtained in central analgesic models where the test drug was comparable or showed less effect than the standard drug paracetamol.

Conclusion

In the experimental study cotton seed Kaṣhayas showed significant anti-nociceptive effects. Highly significant results were obtained in anti-nociceptive models where the test drug was comparable or showed less effect than the standard drug paracetamol.

Figure 1: Karpasa Plant
Figure 2: Karpasa Flower

Figure 3: Gossypium hirsutum L. Seed A. Entire View, B. Longitudinal Section, C. Cross Section

Figure 4: Experimental study

Reference

1. Analgesic [internet], Wikipedia: the free encyclopaedia, [updated on 21 June 2013 at 04:23, cited on 5/7/2013] http://en.wikipedia.org/wiki/Pain.

2. Acharya JT editor. Caraka Samhita of Agnivesa (Ayurveda Dipika, Chakrapanidatta, comme, sanskrit). Varanasi: Chaukhambha Surbharathi Prakashan;2003; p.32.4/8. (Chaukhambha Ayurvijnan Granthmala).

3. P.V.Sharma editor. Suśrutha Samhithā with English translation of text and Dalhaṇa’s commentary along with critical notes. Varanasi: Chaukamba Viswabharati Oriental publishers and Distributers: 2004: Vol-1:p.370. Ch39/7.

4. Sumner, Judith. The Natural History of Medicinal Plants. Timber Press. (2000) p. 17. ISBN 0-88192-483-0. Page no-6.

5. Jadavji Trikamji Acharya (editor). Caraka Samhita (Ayurveda Dipika Commentary of Cakrapanidatta).7th ed. Varanasi: Chaukhambha Surbharati Prakashan; 2008; page no-6.

Manuscript Received	Review Round 1	Review Round 2	Review Round 3	Accepted
2022-10-27	2022-10-29	2022-11-05	2022-11-12	2022-11-19
Conflict of Interest	Funding	Ethical Approval	Plagiarism X-checker	Note
Nil	Nil	Yes	18%

Research Article

Karpasa Beeja

Journal of Ayurveda and Integrated Medical Sciences

Antinociceptive activity of Karpasa Beeja

Abhiram L.^1*

Introduction

Materials and Methods

Observations and Results

Discussion

Conclusion

Reference

Research Article

Karpasa Beeja

Journal of Ayurveda and Integrated Medical Sciences

Antinociceptive activity of Karpasa Beeja

Abhiram L.1*

Introduction

Materials and Methods

Observations and Results

Discussion

Conclusion

Reference

Abhiram L.^1*