ORIGINAL ARTICLE ABSTRACT Objective: To determine the hepatoprotective effect of aqueous extract of Chichorium intybus roots in isoniazid induced hepatotoxicity in adult male mice. Study Design: Experimental study. th th Place and Duration of Study: Study was conducted from 15 of January to 15 of March 2015 at National Institute of Health Sciences (NIH) in collaboration with Riphah Institute of Pharmaceutical Sciences (RIPS). Materials and Methods: Forty four Balb/c albino mice were divided randomly in to two groups, Group A (n=12) a control group and Group B (n=32) ,was given isoniazid 50mg/kg body weight orally once daily along with normal diet and water for 30 days to develop hepatotoxicity. Initially 2 mice from both groups were taken to check the ALT level on day 0. Isoniazid induced hepatotoxicity was confirmed by raised serum ALT levels in a mid-cycle sample of 10 mice from the Group B on day 30 mice (n= 10). After development of hepatotoxicity mice from Group B were further divided into two groups C and D. Group B1 (n=10) were given aqueous extract of Chichorium intybus roots at a dose of 200mg/kg/day and Group B2 (n = 10) at a dose of 400mg/kg/day orally for a duration of 30 days. On day 60 serum ALT of all the mice of Group B1, Group B2 was estimated to determine the hepatoprotective effect of aqueous extract of Chichorium intybus roots in Group C and D. Results: Isoniazid produced severe hepatotoxicity as depicted by raised alanine aminotransferase (ALT) levels. ALT levels were decreased in Group B1 and B2. Conclusion: Aqueous extract of Chichorium intybus roots has significant hepatoprotective effects. Key Words: Chichorium Intybus, Drug Induced Liver Injury (DILI), Hepatoprotoxicity, Isoniazid. 4 and pyrazinamide have hepatotoxic effects. Drug induced liver injury (DILI) caused by the anti- 5 tuberculous drugs varies from 2.0-28%. Drug induced liver injury is clinically manifested by the raised liver enzymes. The most sensitive hepatic injury indicator alanine aminotransferase (ALT) level was measured to see the hepatotoxicity in all 6 groups. is the main antibiotic used for Isoniazid 7 longer duration for the treatment of tuberculosis. Acetyl hydrazine, a metabolite of isoniazid which on 8 bio-activation leads to hepatotoxicity. Plants have been a source of medicinal importance throughout 9 the history. commonly known as Chichorium intybus chicory, has been used as a medication in gastrointestinal and inflammatory diseases, whole plant has got valuable phytochemicals in it however roots contain essential components of therapeutic 10 significance. roots has got Chichorium intybus 1 1 , 1 2 h e p a t o p r o t e c t i v e , a n t i o x i d a n t a n t i - 1 3 1 4 , 1 5 i n f l a m m a t o r y a n t i m i c r o b i a l a n t i - 1 6 , 1 7 1 8 , 1 9 hy p e rg l yc e m i c i m m u n o st i m u l a nt t u m o r 20 inhibitory properties. Traditional medicines and herbs have been used locally in the market and scientific study has not be explored to see the active Introduction Across the globe most of cases of tuberculosis occur 1 due to Mycobacterium tuberculosis. Before the invention of antibiotics tuberculosis was a leading cause of death in both Eastern and Western 2 nations. According to the statistics Pakistan is ranked 4th amongst the multi-drug resistant cases of tuberculosis, approximately 3 million deaths per annum have been recorded with increased 3 frequency of new cases. All major drugs used for the treatment of tuberculosis i.e. isoniazid, rifampicin Hepatoprotective Effect of Aqueous Extract of Chichorium Intybus Roots on Isoniazid Induced Hepatotoxicity 1 2 3 Amanat Ali , Adnan Jehangir , Farhana Ayub JIIMC 2016 Vol. 11, No.3 Correspondence: Dr. Amanat Ali Assistant Professor, Pharmacology HBS Medical and Dental College, Islamabad E-mail: doctoramanatali@gmail.com 1 Department of Pharmacology HBS Medical and Dental College, Islamabad 2 Department of Pharmacology Islamic International Medical College Riphah International University, Islamabad 3 Department of Biochemistry Islamic International Medical College Riphah International University, Islamabad Funding Source: NIL ; Conflict of Interest: NIL Received: Apr 07, 2016; Revised: Jun 12, 2016 Accepted: Aug 06, 2016 Hepatoprotective Effect of Chichorium Intybus 99 principles and phytochemicals. Current research was aimed to see the active principle in the herb and to support it biochemically. Rationale was to explore the scientific evidence of the active ingredients helpful in preventing DILI in patients on antituberculous drugs. The objective of the present study was to explore the hepatoprotective effect of aqueous extract of roots in dose Chichorium intybus d e p e n d e n t m a n n e r o n i s o n i a z i d i n d u c e d hepatotoxicity. Materials and Methods An experimental randomized control study was carried out at Riphah Institute of Pharmaceutical Sciences (RIPS) and National Institute of Health Sciences (NIH), Islamabad. Forty four Balb/c male and healthy albino mice weighting 30-50 grams with normal ALT levels were taken for the study and were acclimatized for one week in the NIH animal house under standard facilities and were given normal diet and water ad libitum. Initially, 44 mice were randomly divided in to two groups, Group A (n=12) which was given normal diet and tap water ad libitum, Group B (n=32) was given isoniazid 50mg/kg body weight orally once daily along with normal diet and water for 30 days to 21,22 develop hepatotoxicity. On day 0 blood samples of two mice from each group were taken through cardiac puncture. After 30 days mid cycle samples of 10 mice from Group B were taken, ALT levels were performed to see establishment of hepatotoxicity. After confirmation of hepatotoxicity mice from Group B were further divided in to two groups, Group B1 n=10 which was given aqueous extract of Chichorium intybus roots at a low dose of 23 200mg/kg/day and Group B2 which was given aqueous extract of Chichorium intybus roots at a high 23 dose of 400mg/kg/day orally for a duration of 30 th days. On termination day i.e. day 60 blood samples were taken from the both experimental Groups B1 and B2 for evaluation of ALT levels. Chichorium intybus was identified by herbarium department, Quaid-e-Azam University, Islamabad. Aqueous extract of Chichorium intybus roots was prepared at RIPS, Islamabad by using fine homogenized powder of dried chicory roots which were mixed with distilled water, the whole solution was boiled for 2 hours and after cooling was sifted through filter paper. The aqueous extract was formed by using vacuum rotary evaporator and was 24 frozen dried. Results were compiled and data was entered into SPSS 17 was used for statistical analysis. Tuckey's multiple comparison test to observe group mean differences. A p-value of <0.05 was considered as statistically significant. Results Serum ALT levels were significantly raised (p<0.01) in Group B treated with isoniazid as compared to Group A. Chichorium intybus roots extract significantly reduced (p<0.01) serum ALT level in Group B1 and Group B2 in comparison to Group B. Table I: Tukey's mul�ple comparisons test between study Groups ANOVA summary F 35.52 P value <0.0001 P value summary **** Are differences among means sta�s�cally significant? (P <0.05) Yes Discussion In the present study mice were treated with isoniazid at 50mg/kg resulted with significant elevation in serum ALT levels. Group B1 and B2 received aqueous extract of Chichorium intybus roots resulted in significant improvement of ALT levels in a dose dependent manner. Our study in accordance with study carried out by El-Sayed et al in 2015 which showed antioxidant activity of Chichorium intybus in 14 CCl4 induced hepatotoxicity. similarly our study is in correlation with another study performed by Atta et al. showing hepatoprotective effect of Chichorium intybus extract when given with methanolic extract 25 of Zinger Officinale. Similar results have been JIIMC 2016 Vol. 11, No.3 Hepatoprotective Effect of Chichorium Intybus 100 found in the study performed by Li et al. on hepatoprotective effect of Chichorium intybus in 26 CCl4 induced hepatotoxicity in rat model. Previously studies have been done on exploring hepatoprotective effect of Chichorium intybus in combination with medical compounds like silymarin and other herbal compounds and extracts. No dose dependent study was done individually on aqueous extract of Chichorium intybus roots extract which guides us about the submaximal, ceiling effect and toxicity. Our study confirms the individual hepatoprotective effect of aqueous extract of Chichorium intybus roots. Further studies are needed to determine molecular mechanism of inulin which is the major active principle of the Chichorium intybus roots. In addition a higher dose and different routes of administration can be tried to see the same effect. 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