6.1 Need for the study:
The liver is largest and important key organ which is concerned with various metabolic processes like drug metabolism, lipid metabolism, carbohydrates and protein metabolism. Various types of liver diseases includes jaundice, acute hepatitis, chronic hepatitis & cirrhosis etc. some important factors responsible for the liver damages are viruses, toxic substances like( allopathic drugs & chemicals), and circulatory disturbances. In the liver disorders the carbohydrates, proteins, lipid as well as drug metabolism may be impaired. Therefore there is impairment in normal healthy life of patient which creates symptoms like anorexia, nausea, vomiting, diarrhea , enlargement of lymph nodes, spleenomegaly, and hepatomegaly etc.
No drug has been developed in modern system of medicines which may stimulates the liver functions, protect it from damage or help in the regenaration of hepatic cells. The only drugs available for treatment of liver disorders are corticosteroids and immunosuppressive agents. But they are having many side effects.
Herbs play a protective role in the management of the various liver disorders. In Indian flora a number of medicinal plants possess hepatoprotective potential. Some of important hepatoprotective polyherbal formulations are Liv-52, Livol, Stimuliv, Hepacure, Hepatoguard and Arogyavardhini.
Hepjaun syrup is one of such popular hepatoprotective product which is combination of 12 natural herbs. Hepjaun is claimed to be useful in hepatitis, jaundice and biliary dysfunction. However, the pharmacological effects need experimental evidence for their actions.
Keeping this in mind and to give scientific validity of it’s therapeutic value as a good hepatoprotective agent, an attempt has been made in this study to examine the hepatoprotective action of proprietary polyherbal product Hepjaun against CCl4 induced hepatotoxicity at different concentration of herbs and modified extraction method.
6.2Review of literature:
1. Subbarao VV and Gupta ML.(1978).Studied the effect of CCl4 and Liv-52 on liver microsomal protein, total protein and nucleic acids. From the study it is clear that prior and subsequent administration of Liv-52prevents the changes caused by CCl4 but Liv-52 alone doesn’t cause any significant change in the protein and nucleic acids.1
2. Sangeeta Shukla Anjana Jadon, Monika Bhadauria and Abhilasha Sharma. (2007). The administration of carbon tetrachloride led to the assimilation of fat in the liver and kidney leading to the increase in acid phosphotase activity. PHF (propriety herbal formulation) therapy showed protective role which might be due to the lysosomal stabilizing property and thus obstruct the rise in acid phosphotase activity. 2
3. NahidTabassum, ShyamS.Agarwal. (2004).Have reported Hepatoprotective activity of Eclipta alba Hassk. against Paracetamol induced hepatocellular damage in mice.Treatment with 50% ethanol extract of E. alba (100&250mg/100g body weight) was found to protect the mice from hepato-toxic action of paracetamol as evidenced by significant reduction in the elevated serum transaminase levels.3
4 Hemalatha K, Nataraj NH, & Kiran A S.(2004). Have reported about the activity of ethanolic extract of dried leaves of Lawsonia alba and its four different crude fractions. The ethanolic extract and other fractions were screened pharmacologically for their hepatoprotective activity against carbon tetrachloride induced hepatotoxicity. The ethanolic extract & butanone fraction exhibited significant activity by lowering serum levels.4
5. Anwar-Ul Hassan Gilani., Khalid Hussain Janbaz.(2006).Studied the effect of aqueous methanol extract of Berberis aristata on paracetamol and CCl4 induced hepatic damage. Post treatment with three successive doses of extract restricted the hepatic damage induced by paracetamol this indicate the crude extract of Berberis aristata exhibits hepatoprotective action. 5
6. Chandan BK, SaxenaAK, Neelam Sharma, et al.(2007).Have studies the effect of different solvents extracts of Aloe barbadensis on CCl4 induced hepatotoxicity. Methanolic and aqueous extracts shows significant hepatoprotective activity. 6
7. Handa SS and Sharma A.(1990). Studied the effect of mehtanolic extract of A. paniculata on
CCl4induced hepatotoxicity. Results suggest that Andrographrolide is a major active
hepatoprotective principle present in A. paniculata.7
8. Devaki T, Shivshangari KS, Ravikumar V & Govindraju P.(2004). Have studied the effect of ethanolic extract of Boerhavia diffusa roots against ethanol induced hepatic damage. Post treatment of B. diffusa extract reverse the hepatotoxic alterations to near normal.8
9. Tasaduq SA, Sing K, Sethi S , SharmaSC, Bedi KL, Singh JJ, et al. (2003).Have studied activity of HP-1 a herbal formulation consisting of Phyllanthus niruri against CCl4 induced toxicity. Results shows that HP1 reverse the leakage of lactate dehydrogenase and glutamate Pyruvate transaminase.9
10. Kale BP, Kothekar MA, Tayade HP, And Jaju JB.(2003). Have studied the hepatoprotective activity of Azadirachta indica aqueous leaf extract on anti- tubercular drug induced hepatotoxicity in Albino rat. Results shows that aqueous leaf extract significantly prevent changes in the serum levels of bilirubin, alanine aminotransferase and alkaline phosphatase.10
11. Jain A, Singhai AK, Dixit VK.(2006). Have studied the effect of ethanol extract of leaves and flavonoids isolated from leaves from Tephrosia pupurea for hepatoprotective activity in Rat by inducing hepatotoxicity with CCl4. Hepatoprotective activity was more in methanolic extract of leave.11
12. Bahar Ahmed, Tanveer Alam, Manoj Varshney, and Shah Alam Khan. (2002).The different extract of A. graveolens and Croton olongifolius were tested for their hepatoprotective activity against CCl4induced toxicity in albino rat. The methanolic extract shows most significant activity. 12
13. Madhulika Singh, Vandana Tiwari, Amita Jain, and Shila Ghoshal.(2005). Have studied the effect of Picroliv isolated from root and rhizomes of Picrorhiza kurroa against Entamoeba hystolytica induced liver damage. Picroliv was found to possess hepatoprotective activity against amoebic liver abscess.13
7 / 6.3The objectives of the present study are:
Preliminary Pharmacognostic Evaluation Of Herbal Drugs used in “Hepjaun” Syrup.
Extraction and formulation of polyherbal product (Hepjaun) by modified method.
Extraction and formulation of polyherbal product (Hepjaun) using different concentration of the herbs.
Standardization of original formulation -Hepjaun syrup.
Pharmacological activity (clinical trials).
MATERIALS AND METHODS:
7.1 Source of data
- Published research papers.
- Review articles from journals.
- Electronic data.
- Library of K.L.E.S’s College of pharmacy.
7 7.2 Method of collection:
All the herbal drugs and additive will be provided by SG Phytopharma Pvt.Ltd., Kolhapur, Maharashtra.
Authentification of the herbs from renowned botanist.
Hepatoprotective activity :
Carbon tetra chloride induced hepatotoxicity model :
Hepatoprotective activity will be carried out using Male Albino rat (150-180gm). The animals are grouped into six of five animals each & maintained on standard diet & water ad libitum.
Group 1.- Serve as control receiving 4% gum acacia 1ml/kg orally for 4 days with 2ml of olive oil given subcutaneously on 2nd &3rd Day.
Group 2 – Serve as toxicant receiving 4% gum acacia 1ml/kg orally for 4 days with 1:1 CCl4 in olive oil. 2ml/kg given subcutaneously on 2nd &3rd Day.
Group 3 – Serve as standard receiving LIV –52 (STD) for 4 days with 1:1 CCl4 in olive oil. 2ml/kg given subcutaneously on 2nd &3rd Day.
Group 4 – Received polyherbal formulation ( Hepjaun syp) at fixed dose of body weight by oral route of administration.for 4 days with 1:1 CCl4 in olive oil. 2ml/kg given subcutaneously on 2nd &3rd Day.
Group 5 & 6 – Received polyherbal formulation prepared by (a) higher concentration & (b)modified method at fixed dose of body weight by oral route of administration.for 4 days with 1:1 CCl4 in olive oil. 2ml/kg given subcutaneously on 2nd &3rd Day respectively.
The method of Handa et al. was used to evaluate CCl4 induced hepatotoxicity. The rats were anaesthetized with ether on 5th day and puncturing the retro-orbital plexus collected blood samples. The serum was separated after coagulation. .Each animal serum was estimated for SGOT, SGPT, SALP and total bilurubin content.
7.3 Does the study require any investigation or interventions to be conducted on the patients or other human/animals? If so, please describe briefly.
The above study requires investigation on Albino rat for hepatoprotective activity and Mice for dose determination of polyherbal formulation respectively
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
Ethical clearance will be obtained from our institution.
8 / LIST OF REFERENCES:
1 Subbarao VV & Gupta ML. Effect of carbon tetrachloride & Liv. 52 on Liver Microsomal protein, Total protein & Nucleic acid. The Indian practioner.1978; 11: 831.
2 Sangeeta Shukla, Anjana Jadon, Monika Bhadauria & Abhilasha Sharma. Prevention of acute carbon tetrachloride induced Hepatic & Renal toxicity in Rats by PHF: A Propriety herbal formulation. International Journal of pharmacology & Biological Sciences. 2007; Vol. 1, No.2 : 71-80.
3 Nahid Tabassum, Agarwal Shyam S. Hepatoprotective activity of Eclipta alba Hassk. Against Paracetamol induced Hepatocellular damage in Mice. J.K. practitioner, 2004; 11(4): 278-280.
4 Hemlatha K, Natraj HN , KiranAS. Hepatoprotective activity of leaves of Lawsonia alba. Indian Journal of Natural products . 2004; Vol. 20; No.4: 14-17.
5 Anwar-Ul_Hassan Gilani, Khalid Hussain Janbaz. Preventive & curative effects of Berberis aristata fruit extract on Paracetamol & CCl4- induced hepatotoxicity. Phytotherapy Research; 2006; Vol. 9; issue 7:489-94.
6 Chandan BK, SaxenaAK,et al. Effect of different extracts of Aloe vera on Hepatotoxicity induced by CCl4 on Rats. Journal of Ethnopharmacology. 2007; Vol. 111; issue 3: 560-66.
7 Handa SS, Sharma A. Hepatoprotective activity of Andrographolide from Andrographis paniculata against CCl4. Indian J. Med. Res.1990 Aug; 92: 276-83.
8 Devaki T, shivshangari KS, RavikumarV & Govindaraju P.Hepatoprotective activity of Boerhavia diffusa on ethanol- induced liver damage in Rats. Journal of Natural Remedies. 2004; Vol.4; No.2: 109-11.
9 Tasaduq SA, Sing K, Sethi S, Sharma SC, Bedi K.L, Singh J, et al. Hepatatocurative &
antioxidant profile of HP-1,a polyherbal phytomedicine. Human and Experimental Toxicology.2003; Vol.22; NO.12: 639-45.
10 Kale BP, Kothekar MA, Tayade HP, And Jaju JB. Effect of aqueous extract of Azadirachta
indica leaves on Hepatotoxicity induced by Antitubercular drugs in Rats. Indian Journal of
Pharmacology 2003;35;177-80.
11 Jain A, Singhai AK, Dixit VK. A comparative study of ethanol extract of Tephrosia purpurea & flavonoid isolated for hepatoprotective activity. Indian Journal of Pharmaceutical Sciences. 2006; Vol.68; Issue 6: 740-43.
12 Bahar Ahmed, Tanveer Alam, Manoj Varshney and Shah Alam Khan. Hepatoprotective activity of two plants belonging to the Apiaceae & the Euphorbiaceae family. Journal of Ethanopharmacology, 2002, March; Vol.79; Issue3: 313-16.
13Madhulika Singh, Vandana Tiwari, Amita Jain, and Shila Ghoshal. Protective activity of picroliv on hepatic amoebiasis associated with CCl4 toxicity. Indian J. Med. Res. 2005 May; 121; PP: 676-82.