Comparative Hylauronidase enzyme activity of Ayurvedic formulation Triphala guggulu

 

Alok Sharma^1*, Dinesh Kumar Mishra², Karunakar Shukla³

¹Research scholar, Uttarakhand Technical University, Dehradun, Uttarakhand

¹Mahakal Institute of Pharmaceutical Studies, Ujjain (M.P.)

²Associate Professor, School of Pharmacy and Technology Management, SVKM’s NMIMS, Shirpur (Maharastra)

³Principal, Royal Institute of Management and Advanced Studies, Ratlam (M.P.)

 

ABSTRACT:

A herbal formulation Triphala Guggulu was prepared based on the method described in Ayurvedic Formulary of India. It contains This triphala guggulu includes Amla (Emblica officinalis), Bibhitaki (Terminalia Bellerica), Haritaki (Terminalia Chebula), Pippali (piper longum), Guggulu (Commiphora mukul) plants. Maximum plant was used in preparation of ayurvedic formulation shows anti inflammatory activities and formulation also use for treatment of inflammation and in arthritis.The aqueous extract of TG exhibited a maximum hyaluronidase enzyme inhibition of 83.90% and marketed formulations TG-1,TG-2  and TG-3 exhibit maximum maximum hyaluronidase enzyme inhibition of 83.86%,83.87% and83.89% respectively. The results of this study suggest that the Hyaluronidase enzyme inhibition of Triphala Guggulu may explain its use in joints related problems like Rheumatoid arthritis, Gout and in inflammation of joints.

 

 

INTRODUCTION:

The term “Ayurveda” combines the Sanskrit words ayur (life) and veda (science or knowledge) which means “The science of life”. Ayurveda was the first holistic system of diagnosis and treatment integrating nutrition, hygiene, rejuvenation and herbal medicine. The most of the Ayurvedic formulation are lacking in their defined quality control parameters and method of its evaluation. The World Health Organization (WHO) has emphasized the need to ensure the quality of medicinal plant products by using modern controlled technique and applying suitable standards. The use of Ayurvedic formulations has led to the sudden increase in the number of Ayurvedic drug manufactures due to the toxicity and side effcts of allopathic medicines.

 

Triphala Guggulu is an Ayurvedic formulation containing Guggulu (Commiphora mukul) is the major ingredient along with Triphala ingredients^1,2,3.

 

The medicinal oleoresins produced by Commiphora mukul were known as myrrh. In Ayurvedic literature, myrrh oleoresins are referred to as guggulu. Guggulu destroys bodily wastes and toxins, boosts metabolism and strength, and rejuvenates the body. Excessive bodily waste and toxins plays a major role in the aetiopathology of rheumatoid arthritis. A study reported the anti-inflamatory activity of crystalline steroids from C. mukul resin^4,5. Guggulu is an exudates obtained from the plant Commiphora mukul. Preparations having the exudates as main effective ingredient are known as Guggulu. There are five different varieties of Guggulu described in the Ayurvedic texts. However two of the varieties, namely, Mahisaksha and Kanaka Guggulu are usually preferred for medicinal preparations. Mahisaksha Guggulu is dark greenish brown and Kanaka Guggulu is yellowish brown in color⁶.

Triphala guggulu is a polyherbal ayurvedic formulation.This medicines is guggulu kalp and contain guggulu more than other ingredients use to make this medicines. As the name itself suggests that medicine contains triphala and guggulu .triphala improves digestion, cure constipations and detoxifies bodies. Guggulu gives relief in inflammation and pain.Triphala guggulu is indicated in low metabolism, obesity, piles, fistula, inflammation and gout etc^{7, 8, 9}.

 

The formulation taken for the present study is as compiled in Sarangdhara samhita This triphala guggulu includes Amla (Emblica officinalis), Bibhitaki (Terminalia Bellerica), Haritaki (Terminalia Chebula), Pippali (piper longum), Guggulu (Commiphora mukul) plants. Phenolic compounds of methanolic extract of Amla (Emblica officinalis) may serve as a potential herbal drug for amelioration of acute inflammation due to their modulatory action on free radicals^{10, 11}. Haritaki (Terminalia Chebula) used in the treatment of arthritic disorders and suggest that radical quenching may be one of the mechanisms for its anti-inflammatory activity  and also show antioxidant activity^12,13,14. Pippali (piper longum) dried fruit's oil was taken for the study. The results obtained indicate that the Piper longum oil had significant antiinflammatory activity in rats^15,16,17. Guggulu (Commiphora mukul) has been extensively studied in inflammation and pain in bones, joints, muscles and related connective tissues^18-22.

 

The Components of Triphala were also evaluated for their inhibitory activities on hyaluronidase and collagenase. This is the fi rst report to show that the T2 component of Triphala (T. chebula) is a highly potent hyaluronidase and collagenase inhibitor²³.

 

MATERIAL METHODS:

Assay was developed to investigation this hypothesis. First, we tested the effects of the Triphala guggulu home made formulation (TG) and its marketed formulations TG-1, TG-2, TG-3) on the activity of hyaluronidase, an enzyme that destroys the hyaluronic acid (HA) backbone of cartilage matrix. Indeed, the serum level of HA is a reliable biomarker of Rheumatoid arthritis progression²⁴.

 

Triphala Guggulu Preparation:

Wash, dry and powder the ingredients Amla (Emblica officinalis), Bibhitaki (Terminalia Bellerica), Haritaki (Terminalia Chebula), Pippali (piper longum), of the formulation composition separately and pass through sieve number 85, weigh them separately in the required quantities and mix. Crush weighed quantity of Guggulu-suddha, add fine powder of other mixed ingredients to it and pound well and mix properly. Add Ghrita to smooth the progress of the pounding and carry on pounding till a semi-solid consistently mixed mass of suitable plasticity is obtained. for 8 to 10 h. This Roll the Vatis on flat surface to round them by circular motion of palm covered with a glove and smeared with Ghrita or use suitable mechanical device (Table 1). Dry the rounded Vat is in a tray dryer at not more than a temperature 600ºC. formulation is known as TG. A sample of this formulation was used for this study. Three marketed preparation of Triphala Guggulu also use for comparative study of Triphala Guggulu. This formulation was known as TG-1, TG-2  and TG-3. A sample of this marketed formulation also used for study.

 

Preparation of Aqueous extract:

Aqueous extracts: Triphala guggulu TG were solubilized in distilled water (20–50 mg/ml) by limited autoclaving (5 lbpressure, 8 min) to yield extracts. Same method was also used for marketed preparation TG-1,TG-2 and TG-3 respectively.

 

Hyaluronidase Assay:

Hyaluronidase was assayed by a highly sensitive spectrophotometric method, based on precipitation of HA with cetylpyridinium chloride, which is used for high throughput screening for hyaluronidase inhibitors²⁵. Enzyme (800 U/ml) and HA substrate (0.40 mg/ml) were incubated at 37°C for 1 h. Enzyme activity was measured by monitoring the percentage of undigested HA substrate in the cetylpyridinium chloride precipitate at absorbance 415nm after the enzyme reaction (Table 2).

 

Table-1 Composition of Triphala Guggulu (Ayurvedic formulary of India)

 

Table-2 Percent inhibition of hyaluronidase by TG formulations

	Aqueous extracts
Test sample	Without TG	With TG	With TG1	With TG2	With TG3
Hyaluronic acid (HA)	0.489± 0.033	0.538± 0.040	0.535± 0.020	0.536± 0.018	0.539± 0.016
HA + enzyme	0.057± 0.031	0.514± 0.039	0.511± 0.032	0.512± 0.032	0.515± 0.031
Enzyme activity	88.34%	4.44%	4.48%	4.47%	4.45%
% enzyme inhibition		83.9% (88.34–4.44)	83.86% (88.34–4.48)	83.87% (88.34–4.47)	83.89% (88.34–4.45)

Triphala guggulu (TG), expressed as mean ± SD of 3 experiments

Formulae were used for calculations:

% Enzyme activity =

(100%) – {A415 nm of HA + enzyme/A415 nm of HA x 100}

 

RESULT AND DISCUSSION:

Triphala guggulu contain Triphala and Guggulu. Triphala synergized with the guggulu resin in enhanced the hyaluronidase activities of the TG, TG-1, TG-2  and TG-3 formulations.

 

Percentage inhibition of hyaluronidase enzyme for Triphala guggulu formulation found to be for TG 83.90% and for TG-1, TG-2 and TG-3 are 83.86%, 83.87% and 83.89% respectively. Maximum hyaluronidase enzyme inhibition found in TG and minimum hyaluronidase enzyme inhibition found in TG-3. In all formulation hyaluronidase enzyme inhibition found to be close to 83%.

 

This triphala guggulu includes Amla (Emblica officinalis), Bibhitaki (Terminalia Bellerica), Haritaki (Terminalia Chebula), Pippali (piper longum), Guggulu (Commiphora mukul) plants. Phenolic compounds of methanolic extract of Amla (Emblica officinalis) it serve as a potential herbal drug for acute inflammation due to their modulatory action on free radicals. Haritaki (Terminalia Chebula) used in the treatment of arthritic disorders Piper longum oil had significant antiinflammatory activity in rats. Guggulu (Commiphora mukul) has been extensively studied in inflammation and pain in bones, joints, muscles and related connective tissues.

 

This result become important when we used costly injections of hyaluronic acid into the synovial cavity is administered to patients whose suffer from chronic arthritis. It may become a alternative.

 

CONCLUSION:

Percentage inhibition of hyaluronidase enzyme study on Triphala Gugglu In house and marketed formulations are studied. The result shows that triphala guggulu may be use in certain type of diseases like joints pain, inflammation, Rheumatoid Arthritis. Threfore, it has been concluded that the formulation prepared by traditional method reported in Ayurvedic Pharmacopoeia is the best formulation in respect of that activity.

 

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Accepted on 24.12.2017        © RJPT All right reserved

Research J. Pharm. &Tech 2018; 11(2):463-465.