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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 16  |  Issue : 3  |  Page : 178-184

Evaluation of Shonitasthapana karma of Yashtimadhu (Glycyrrhiza glabra linn.): An experimental study


Department of Dravyaguna Vigyana, National Institute of Ayurveda, Jaipur, Rajasthan, India

Date of Submission10-Mar-2021
Date of Decision29-Sep-2021
Date of Acceptance27-Dec-2021
Date of Web Publication28-Sep-2022

Correspondence Address:
Richa Khandelwal
4/144, Vidhyadhar Nagar, Jaipur, Rajasthan
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/joa.joa_67_21

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  Abstract 


Introduction: Many pharmacological actions have been mentioned in Ayurveda literature. However, there is no equality in understanding of these actions because of different commentator's view, so this study has been planned to reevaluate one such type of pharmacological action, shonita sthapana by an experimental method by seeing the effect of yashtimadhu on various blood parameters and coagulation profile in normal albino Wistar rats and Swiss albino mice. Methods: This study was conducted in two parts. The first experiment was done to evaluate the effect of madhuka on 18 normal albino Wistar rats by dividing three groups of 6 rats and giving carboxymethylcellulose (CMC) solution, 360 mg/kg madhuka powder, and 180 mg/kg madhuka powder, respectively. The second experiment was done to evaluate the effect of madhuka in 18 swiss albino mice by dividing them in three groups of 6 mice and giving CMC solution, 360 mg/kg madhuka powder and 180 mg/kg madhuka powder, respectively, for 23 days and aspirin at 600 mg/kg b.w. for last 7 days along with vehicle and drug. Different hematology analysis and coagulation profile were evaluated. Results: There is no any significant result was found for any parameter in the experiment. Conclusion: Yashtimadhu is not having any effect on blood parameters and coagulation profile in present experimental model.

Keywords: Blood components/parameters, coagulation, experimental study, Glycyrrhiza glabra linn, shonita-sthapana karma, yashtimadhu


How to cite this article:
Khandelwal R, Nathani S, Sharma G. Evaluation of Shonitasthapana karma of Yashtimadhu (Glycyrrhiza glabra linn.): An experimental study. J Ayurveda 2022;16:178-84

How to cite this URL:
Khandelwal R, Nathani S, Sharma G. Evaluation of Shonitasthapana karma of Yashtimadhu (Glycyrrhiza glabra linn.): An experimental study. J Ayurveda [serial online] 2022 [cited 2022 Nov 29];16:178-84. Available from: http://www.journayu.in/text.asp?2022/16/3/178/357304




  Introduction Top


Ayurveda is the age-old science and has very enormous ancient literature of Sanskrit language. Many pharmacological actions are mentioned in ayurveda classics i.e., Deepana (appetite increasing), pachana (increases digestive fire), jeevaneeya (vitaliser), bhedana (cholagogue purgation), rechana (purgation), etc., A maximum of them were explained and understand well. But some of them are not easily understandable because different commentators gave their comments with different understandings. Hence, there are so much dissimilarities and confusion among vaidyas about these terminologies. Some examples of these type of terms are shonita-sthapana, vedana sthapana, etc., It is a need of time to reevaluate our principals by different experimental and clinical setups for easy understanding.

Shonita-sthapana term was reevaluated in this study by experimental methods. Shonita sthapana mahakashaya was mentioned in charaka samhita in chapter 4, shadvirechan shatashriteeya adhyaya.[1] The word shonita-sthapana is described by different commentators.

Chakrapani[2] and Shivadasa sena[3] interpret this as action which clears the impurities of blood and keep it in its natural form. Yogindranatha sena[3] and Ḍalhana[4] interpret it as the action which is related to coagulation. Gangadhara comprises both views in his statement. However, according to Indu, shonita-sthapana is which increases the blood and also keeps it in its natural form.[5] On the basis of these statements of commentators, it can be said that shonita-sthapana covers three areas – Rakta-prasadana – which clear the impurities or vitiation in the blood, Rakta-stambhana – which stop the bleeding conditions, Rakta-vardhana – which increases the blood.[6]

There are ten drugs mentioned in shonitasthapana mahakashaya – madhu, madhuka, rudhir, mocharasa, mritkapal, lodhra, gairika, priyangu, sharkara, and laja.[7]

Madhuka (Glycyrrhiza glabra Linn.) is the multifaceted drug which is indicated in various bleeding conditions as well as for various anabolic processes in the body. Yashtimadhu (G. glabra Linn.) is a very profoundly used medicine in Ayurveda classics. It is mentioned in 11 mahakashaya (a group of 10 drugs having the same pharmacological action) in charaka samhita among fifty mahakashaya.[8] It is mentioned in madhura skandha (group of drugs having sweet taste) also.[9] Among 36 groups of drugs (gana) of Sushruta samhita, it is included in 8 groups (Gana).[10] In Ashtanga Hridaya, yashtimadhu (G. glabra Linn.) has been included in ten ganas out of thirty-three Ganas.[11] It expresses that it is a broad-spectrum drug that is used very extensively in Ayurveda pharmacology.

So, this study was carried out to reevaluate the term shonita-sthapana and its extents with madhuka (G. glabra Linn.) as prototype drug.


  Materials and Methods Top


An experimental study was conducted in Institute of Biomedical and Industrial Research, Vidyadhar Nagar, Jaipur (CPCSEA Registration No: 1737/PO/Rc/S/14/CPCSEA) after approval from the Institutional Ethics Committee with IAEC Approval No: IBIR/IAEC/2015/I/4.

Test sample

Madhuka (G. glabra Linn.) dried roots were purchased from Jaipur crude drug market and authentication was done from Raw Material Herbarium and Museum, Delhi (RHMD) of National Institute of Science Communication and Information Resources (NISCAIR) with Ref. no. NISCAIR/RHMD/Consult/2016/2977-04. Powder was made by the grinder.

Chemicals and equipment

Chemicals being used throughout the experimentation are picric acid, distilled water, normal saline. Equipment which is used for the experimentation is polypropylene cages, water bottle, anesthetic chambers, syringe, weighing machine, etc.

Experimental animals

18 wistar strain albino rats of either sex weighing 150 ± 20 g and 18 swiss albino mice of either sex weighing (25 ± 5 g) were used for this experiment.

Housing and feeding conditions

The temperature in the experimental animal room was maintained at 22°C (±3°C), and relative humidity was maintained at 50%–60% during room cleaning. Lighting was artificial, the sequence of 12 h light and 12 h dark. For feeding, conventional laboratory diets of Gulmohar brand animal feed manufactured by Lipton India Limited were used with an unlimited supply of drinking water.

Methodology and grouping

This study was conducted in two parts.

A. Evaluation of madhuka (Glycyrrhiza glabra Linn.) on Blood Components and Blood Coagulation by Rat Tail Cut Method after oral administration:

Group design

18 adult albino Wistar rats were divided into three groups having six rats in each. These groups received different treatment in the following manner:

  1. Group 1 (Control Group): 6 rats of group 1, treated with 1% carboxymethylcellulose (CMC) solution for 1 month
  2. Group 2: 6 Rats of group 2, treated with Madhuka (G. glabra Linn.) (360 mg/kg twice a day) for 1 month
  3. Group 3: 6 Rats of group 3, treated with Madhuka (G. glabra Linn.) (180 mg/kg twice a day) for 1 month


Dose calculation

According to API, dose of madhuka, (G. glabra Linn.) is 2–4 g. On the basis of Paget and Barners (1964) formula,[12] this adult dose is converted into animal dose (minimum and maximum dose). This is applicable for both experiments (A and B).

Experimental procedure

The adult wistar rats (100 ± 20 g) were randomly divided into three groups of six rats per group. After 7 days acclimatization period, test sample at dose (360 mg/kg and 180 mg/kg) and CMC solution had been administer by gavage using an oral feeding needle. Animals had been fasted prior to dosing.

The rats were slightly anaesthetized. Bleeding time and clotting time were measured by cutting off the rat's tail tip (5 mm), and blood for hematology analysis (hemoglobin [Hb], white blood cell [WBC], red blood cell [RBC], neutrophils, lymphocytes, eosinophils, monocytes, basophils, platelet count, prothrombin time, and activated partial thromboplastin time) had been collected from orbital puncture method of blood collection in rats.

B. Evaluation of Test Drug on Blood Components and Blood Coagulation by Rat Tail Cut Method after Aspirin Administration in Swiss Albino mice.

Oral administration of aspirin at the dose of 600 mg/kg body weight in swiss albino mice for 7 days had been shown marked changes in various blood components.[13] That's why aspirin was used to derange blood parameters, and the effect of madhuka (G. glabra Linn.) was evaluated on these deranged blood parameters.

Group design

18 adult swiss albino mice were divided into three groups having six mice in each. These groups received different treatment in the following manner:

  1. Group 1 (Control group): 6 mice of group 1 treated with 1% CMC solution\for 23 days and after 24th day to 30th day aspirin 600 mg/kg body weight with CMC solution
  2. Group 2: 6 mice of group 2 treated with Madhuka (G. glabra Linn.) (360 mg/kg twice a day) for 23 days and after 24th day to 30th day aspirin 600 mg/kg body weight with test sample
  3. Group 3: 6 mice of group 3 treated with Madhuka (G. glabra Linn.) (180 mg/kg twice a day) for 23 days and after 24th day to 30th day aspirin 600 mg/kg body weight with test sample.


Experimental procedure

The adult swiss albino mice (25 ± 5 g) were randomly divided into three groups of six mice per group. After 7 days acclimatization period test sample at dose (360 mg/kg, 180 mg/kg) and 1% CMC solution had been administered for 23 days and after 24th day to 30th day aspirin 600 mg/kg body weight and test sample by gavage using an oral feeding needle. Animals had been fasted prior to dosing.

The mice were slightly anesthetized. Bleeding time and clotting time was measured by cutting off the rat's tail tip (5 mm). And blood for hematology analysis (Hb, WBC, RBC, neutrophils, lymphocytes, eosinophils, monocytes, basophils, platelet count, prothrombin time, and activated partial thromboplastin time) had been collected from cardiac puncher.

Bleeding time was determined using a modified Duke method,[14] and clotting time was determined by the method adopted by Shrivastava B. K et al.[15] in both experiments.

Statistical analysis

The results were expressed as mean ± standard error of mean Comparison between the treatment groups and control were performed by analysis of variance. In all tests the criterion for statistical significance was P < 0.05.


  Results Top


In this study, Evaluation of madhuka was done on Blood Components and Blood Coagulation by Rat tail cut method after oral administration and local application. Results were shown in tabular form:

A. Evaluation of test drug on Blood Components and Blood Coagulation by Rat tail cut Method after oral administration.

After 30 days administration of Madhuka (G. glabra Linn.) powder and CMC, various blood components and coagulation profile are evaluated and it is observed that although there is some percent difference in between control group and test group, statistically insignificant changes were found in all parameters after comparison with Negative control (Group A) and in between test group (Group B, Group C) as shown in [Table 1] and [Table 2].
Table 1: Effect of madhuka on blood components in Group A, B and C

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Table 2: Effect of madhuka on blood coagulation profile in Group A, B and C

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According to [Table 3], which is showing difference between various blood components and coagulation profile in between Group B and Group C with respect to Group A, there is no any significant changes [also showing in [Graph 1]]. It suggests that there is no any dose response after increasing the dose from 180 mg/kg to 360 mg/kg.
Table 3: Difference in blood components and coagulation profile in Group B and Group C w.r.t. Group A

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B. Evaluation of Test Drug on Blood Components and Blood Coagulation by Rat tail cut Method after Aspirin Administration in Swiss Albino Mice.

After 30 days administration of Madhuka (G. glabra Linn.) powder and CMC and aspirin administration in dose of 600 mg/kg on last 7 days, various blood components and coagulation profile are evaluated, and it is observed that although there is some percent difference in between control group and test group but statistically insignificant changes were found in all parameters after comparison with Negative control (Group A) and in between test group (Group B, Group C) as shown in [Table 4] and [Table 5].
Table 4: Effect of madhuka in Group A, B and C on blood components after aspirin administration

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Table 5: Effect of madhuka in Group A, B and C on blood coagulation profile after aspirin administration

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According to [Table 6], which is showing difference between various hematological parameters and coagulation profile in Group B and Group C with respect to Group A which is baseline, there are no any significant changes [also shown in [Graph 2]]. It is suggesting that there is no any dose response after increasing the dose from 180 mg/kg to 360 mg/kg.
Table 6: Difference in blood components and coagulation profile in Group B and Group C w.r.t. Group A

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  Discussion Top


In first experiment, test drug was evaluated for its effect on blood components and blood Coagulation by Rat tail cut Method after oral administration. It was observed that although there were some percent difference in between control group and test group but statistically no any significant changes found in any parameters after comparison with Negative control (Group A) and in between test group (Group B, Group C) were found.

In the second experiment, test drug was evaluated for its effect on Blood Component and coagulation profile after aspirin administration (600 mg/kg for 7 days) in Swiss albino mice. And it was observed that although there were some percent difference in between the control group and test group but statistically no significant changes in any parameters after comparison with Negative control (Group A) and in between test group (Group B, Group C) was found.

There is no any significant result was found in both experiments.

On the basis of above results obtained, it is suggested that the test drug madhuka (G. glabra Linn.) has no significant activity on blood components and coagulation profile in normal albino Wistar rats and swiss albino mice with deranged blood parameters.

But Acharya Charaka included this in shonita-sthapana mahakashaya, then the question arises 'How does madhuka (G. glabra Linn.) work as shonita-sthapana?

Shonita-sthapana karma, as previously said, is combined form of Rakta-prasadana, Rakta-stambhana and Rakta-vardhana action. Although some classical references have been found in which madhuka (G. glabra Linn.) has been used for prasadana and stambhana karma, internally and externally both. Here, we trying to evaluate this by experimental model. Another study has been done to evaluate the effect of yashtimadhu (G. glabra Linn.) infusion and powder on bleeding time with 24 wistar strain albino rats by rat tail cut method. Here also no any significant result was found.[16]

Properties of madhuka (G. glabra Linn.) are madhura (sweet) and tikta (bitter) rasa (taste), guru (heavy) and snigdha (untous) guna (poperties) and madhura (sweet) vipaka[17] and maximum of nighanṭus quoted its raktadoshahara (useful in vitiation of blood) action.[18],[19],[20],[21] They all are suggestive of rakta-prasadana property. Some references are found about rakta-stambhana property like Sushruta and Vagabhaṭṭa have been used madhuka (G. glabra Linn.) for avacurnana (sprinkling of powder) in atisruta rakta avastha (excessive hemorrhage)[22] and this plant is also used in various bleeding conditions internally and externally both. Thus, rasapanchaka and aptopadesha (classical description) are suggestive of our hypothesis.

However, in experimental study, effect of madhuka (G. glabra Linn.) (in powder form) on blood components and coagulation profile was found insignificant statistically.

Some Researchers have demonstrated the ability of liquorice to treat adrenal insufficiency as well as stress intolerance via enhancement of the adrenal cortisol response. Through its inhibitory effect on 11-β-HSD-1, the enzyme responsible for inactivating cortisol, Glycyrrhiza increases activity at mineralocorticoid receptors and promote potassium excretion and sodium and water retention. This may be therapeutic in cases of hyperkalemia and hypoaldosteronism.[23]

Animal studies have also shown Glycyrrhiza to decrease elevated serum cortisol, ACTH, and potassium, while increasing renin, aldosterone, and sodium concentrations.[24]

These effects suggest direct activity on the adrenal-renal-pituitary axis. This aldosterone-like action may cause fluid retention and elevation of blood pressure in some sensitive individuals, but improve symptoms of mineralocorticoid insufficiency. Due to its aldosterone-like effects, Glycyrrhiza is also appropriate for those patients with adrenal insufficiency and related hypotension.[24]

On the basis of above discussion, it can be said that madhuka (G. glabra Linn.) improves blood plasma volume in some therapeutic conditions, so it can come under the third category, rakta-vardhak.

Some researches had shown the effect of G. glabra Linn. on some haematological parameters but when used in extract form[25] or infusion and tea form.[26]

But for this study, madhuka (G. glabra Linn.) was use powder form.

As for the rakta-prasadana (which clear the impurities or vitiation in the blood) and rakta-stambhana (which stop the bleeding conditions) more experiments and research work are needed to reach any conclusion. As Acharya Charaka said “siddhanto nama sa yah parikṣhakairbahuvidham parikṣya hetubhishca sadhayitva sthapayate nirnayah[27] means the demonstrated truth established after several examinations and reasonings is known as siddhant. When we talk about shonita-sthapana, it requires repeated examinations. In this case when examining from aptopadesha (classical references) and anumana (according to rasapancaka) pramana, it is in the favor of our hypothesis, but on performing study in rat model (pratyakṣa pramana), no results were found. This may be due to the reason that this experiment was performed in rat model. Animal experimentation has some limitations. Human and animal physiology is somewhat different. That's why bahuvidha pariksha (repeated examinations) is required.

Moreover, as said by Aruṇadatta, in the commentary of Aṣṭanga Hrdaya, sutra sthana 11/24–25, vriddhi and kshaya of dosha, dhatu, and mala is assessed by the increase or decrease of guna-karma of respective dosha, dhatu, and mala.[28] These specific guna-karma of dhatu after vardhana are more subjective, and their assessment in experimental study is quite difficult. That's why these parameters should be assessed by clinical trials.


  Conclusion Top


Shonita-sthapana karma has been included three parameters, rakta prasadana (Blood purification), rakta stambhana (blood coagulation) and rakta vardhana (increase in decreased blood components). However, in the present study, there is no any significant effect of yashtimashu (G. glabra Linn.) was found. So, on the basis of above experiment it can be concluded that there is no any significant activity of madhuka (G. glabra Linn.) On blood components and coagulation profile in normal wistar strain albino rats and swiss albino mice with deranged blood parameters in the present experiment. It is suggested that this experiment should be performed in different setups in different models as well as in clinical setups to know the different extents of shonita-sthapana karma.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.





 
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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]



 

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