Hepatoprotective Effect of Pomegranate: Histological Study

Hepatoprotective Effect of pomegranate tree Histological Study pilot ARTICLEA HISTOLOGICAL STUDY OF HEPATOPROTECTIVE EFFECT OF POMEGRANATE, IN minocycline TREATED GUINEA PIGSDr. M. Nadeem SiddiquiDr. Sarwath FatimeeDr. Hemant KumarDr. Bashir A. SheikhABSTRACTObjective To determine the hepatoprotective effectuate of pomegranate on drug induced autoimmune hepatitis produced by minocycline. visible and Method This experimental study was conducted in the department of anatomy, basic medical sciences institute, Jinnah post-graduate medical centre, Karachi. 30 adult male guinea pigs taken and dissever in three chemical gatherings, A, B and C, respectively with each gathering having 10 sensuals, sepa give awaye A served as control, Group B was treated with minocycline 0.02mg/Gram bole burthen and Group C was given the minocycline along with pomegranate in the dose of 0.9 mg/ Gram frame weight. On the completion of study, the animals were sacrificed and liver were removed for the histologic mental testing beneath light microscopy in haematoxylin eosin staining.Results The HE stained slides were studied and showed the regulation architecture of liver in group A animals, misrepresent sit of liverwort corduroys with marked infiltration of seditious cells in group B animals, and less(prenominal) altered pattern of hepatic cord with less infiltration of inflammatory cells in group C animals approximately confusable architecture to control group.Conclusion Use of pomegranate with minocycline cigarette prevent hepatic damage in particular the autoimmune hepatitis.KEY spoken communication Minocycline, Pomegranate , hepatotoxicity , autoimmune hepatitis.INTRODUCTIONdo drugss are the most common clear of liver injury. More than 900 drugs toxins and herbs have been reported to ca utilization liver injury.1 Drug induced autoimmune hepatitis (DIAH) has been reported to be caused by several drugs.2 autoimmune hepatitis is a chronic inflammatory diseas e of the liver.3 marked by big infiltration of inflammatory cells.4it is an acute and potentially severe side effect, particularly often reported after the long term use of minocycline.5 A significant proportion of patients with autoimmune hepatitis have drug induced hepatitis, generally because of this drug.6 Minocycline, a semi synthetic, broad spectrum antimicrobial tetracycline available since 1972 is wide prescribed as systemic antibiotic for acne vulgaris which is given for a protracted period ( month to years).6 fodders rich in natural antioxidants have been proposed as a tool to prevent and cure liver damage.7 The pomegranate is bingle of the important dietary sources, that exhibits strong anti oxidant and anti-inflammatory properties.8 Its main constitute, ellagic acid, have been proven to antagonize in vitro to inhibit the pro-inflammatory mechanisms.9 existent METHODThis experimental study was conducted for 8 weeks by taking 30 adult male guinea pigs of 450-650 wei ght at animal stick out of BMSI, JPMC, Karachi and divided them into three main groups i.e., A,B and C that comprises 10 animals each and receiving bar laboratory diet ad labitum. Group A served as control, group B was given minocycline 0.02 mg /G body weight / day orally based on human consumption for the era of one year, while group C was given minocycline in the same dose along with pomegranate 0.9 mg /G body weight /day orally. At the end of the experimental period, all the animals were sacrificed under ether anesthesia and liver was removed to be fixed in 10% formalin for 24-48 hours. The tissue samples taken from the organ were then keep in ascending strength of alcohol, cleared in xylene, infiltrated and embedded in paraffin wax. Tissue blocks were made and 4-5 m thick classs were obtained by incisive them with microtome. The sections then mounted on glass slides and were stained with haematoxylin and eosin. The morphological changes in typical architecture of liver lo bule were observed under light microscope with the support of 8X ocular, 40X objectives and graded according to the infiltration of inflammatory cells within the three zones of hepatic lobule 8. The grading was done as follows sucker O (normal N) no infiltration of infiltration of inflammatory cells in any zone.Grade I (mild M+) infiltration of inflammatory cells within zone IGrade II (moderate M++) infiltration of inflammatory cells within zone IIGrade tercet (marked M+++) infiltration of inflammatory cells within zone IIIRESULTSThe present study was design to observe the minocycline treated animal and pomegranate protected animal in guinea pigs. Observations and results of microscopic examination were compared with control animal.The minocycline treated group B animal showed the distorted pattern of hepatic cords, dilated underlying vein with marked congestion, and some fruitless hepatocytes with piknotic nuclei. Areas of marked infiltration of inflammatory cells especiall y lymphocytes and plasma cells observed in zone III of hepatic lobule.The microscopic examination of pomegranate and minocycline treated group C animals showed the architecture of hepatic lobule in proportion to control group A that is less distorted of hepatic cords and less dilated central vein, while infiltration of inflammatory cells is markedly reduced and limited to zone I when compared to group B animalsn figure of animalsN normalM+ Mild.M+++ MarkedDISCUSSIONThe histological examination of hepatic lobules in haemotoxylin and eosin stained sections of control group A showed normal architecture as set forth by novel et al.10,11In the minocycline treated animals of group B the architecture of hepatic lobule has markedly changed when compared to group A. It showed the ir constant distorted pattern of hepatic cords, cast up in vascularity and irregular shape of hepatocytes with pyknotic nuclei. This is in agreement with Lawrenson et al. who described the comparative changes in hepatic architecture after the use of minocycline that showed the necrotic and inflammatory changes with distorted pattern of hepatic cords.12 This is also reason by Kumar et al. who also showed the necrotic changes and plasma cell infiltration with distorted pattern of hepatic cords in autoimmune hepatitis.13Group C animals treated with pomegranate and minocycline simultaneously demonstrate the protective do on hepatic architecture. The hepatic cords showed regular arrangement of and the hepatocytes were normal in shape, while the hepatic lobules showed less accumulation of inflammatory cells when compared to Group B animals. This is in accordance with the study of Novarro et al. who proved that after the intercession of pomegranate in hepatoma the pattern of distorted hepatic cords become normal.14 This is also in concurrence with the study of Middha et al. who concluded that function of pomegranate, ultimately improve the hepatic architecture by providing the protection ag ainst oxidative damage.15CONCLUSIONFrom the results of this study, it can be concluded that pomegranate has proved to be highly effective in preventing the hepatic damage produced by minocycline particularly minocycline induced autoimmune hepatitis. So use of pomegranate should be promoted in daily life, particularly in those patients who are on long term minocycline treatment.Figure 1 HE stained 4 micron thick sections of minocycline treated liver, showing dilated central vein (CV), distorted hepatic cords with numerous pyknotic nuclei (PN) and marked infiltration of inflammatory cells (IC). (Photomicrograph X400).Figure 2 HE stained section 4 micron thick of pomegranate protected in minocycline treated liver, showing less dilated central vein (CV), more regular hepatic cords (HC) with normal nuclei and less infiltration of inflammatory cells (IC). (Photomicrograph X 400).REFERENCESMehta N, Ozick L. Drug-induced hepatotoxicity.Mar 28 2008, From http// emedicine.medscape.com/article /169814.Bjrnsson E, Talwalkar J, Treeprasertsuk S, Kamath PS, Takahashi N, Sanderson S, Neuhauser M, Lindor K. Drug-induced autoimmune hepatitis clinical characteristics and prognosis. Hepatology. 2010 Jun51(6)2040-8.Manns MP and Vogel A. Autoimmune Hepatitis, From Mechanisms to Therapy.HEPATOLOGY200643S132-S144.Aldenhoven M, van Enk JG, Avis WA. Czaja AJ. Minocycline-induced autoimmune hepatitis. Ned Tijdschr Geneeskd. 2013157(3)A5465.Drug induced hepatitis with autoimmune features during minocycline therapy. 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From http//dx.doi.org/10.1155/2013/656172.