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Abstracts JOURNAL OF CLINICAL AND EXPERIMENTAL HEPATOLOGY
21st Annual Conference of Indian National Associationfor the Study of Liver (INASL), March 22–24, 2013
Hyderabad International Convention Centre, Hyderabad, India
MOLECULAR AND CELLULAR BIOLOGY
IMPACT OF CHRONIC ETHANOL ON THEHORMONAL ACTIVITY OF ADIPOSE TISSUE
Venkata Harini Kema, Nishank Reddy Mojerla,Raghava Jagadeeesh Salaka, Palash Mandal
Department of Biological Sciences, Birla Institute of Technology andScience Pilani, Hyderabad Campus, Hyderabad, Andhra Pradesh, India
Background and Objectives: Consumption of alcoholhas been in existence in the world for many centuries. Everyyear the use of alcohol kills 2.5 million people accountingfor almost 4% of the deaths in the world. Progression of al-coholic liver disease is a multifactorial process involvinga number of genetic, nutritional and environmental fac-tors. Alcohol is metabolized via alcohol dehydrogenasepathway, the microsomal ethanol-oxidizing system involv-ing the Cytochrome P450 2E1 (CYP2E1) enzyme, and cat-alase pathway. Chronic ethanol consumption activatesCYP2E1 enzyme resulting in reactive oxygen species forma-tion leading to oxidative stress and endoplasmic reticulumstress. Excessive oxidative stress results in membrane lipidperoxidation and formation of toxic aldehydes. We hy-pothesize that ethanol induced oxidative and endoplasmicreticulum stress disrupt the secretion of adipokines fromadipose tissue and adipokines are known to play a majorrole in the progression of alcoholic liver disease.Objectives: 1. Studying the over expression and inhibitionof CYP2E1 on ethanol induced oxidative stress and on adi-pokine levels. 2. Investigating the effects of ethanol on theendoplasmic reticulum (ER) function and the influence ofER stress on the posttranslational modifications of adipo-kines. 3. Identification of protein adducts formed by lipidperoxidation products and investigating the resistance tothe metabolic actions of adipokines by hepatic nonparen-chymal cells.Results: Several passages of 3T3L1 cell line and RAW 264.7cell line were established. The cell lines were exposed todifferent concentrations of ethanol. Preliminary data per-taining to the oxidative stress markers was obtained.Quantitative real time PCR studies of CYP2E1, IL-6, leptin,resistin genes are being carried out.Conclusion:Understanding the molecular mechanisms ofethanol mediated liver injury will aid in identification ofnew integrative approaches as it relates to alcoholic liver
© 2013, INASL Journal of Clinical a
injury and provide potential new directions to developtherapeutic target intervention.
Corresponding author. Palash MandalE-mail: [email protected]
DIFFERENTIAL EXPRESSION OF HUMANTOLL-LIKE RECEPTOR 2 AND 4 GENES INHEPATITIS B VIRUS INFECTION
Veena Shravanthi Gelli,1 Veena Shravanthi Gelli,1
Aparna Jakkampudi,1 Ramya Kota,1
Rathindra Mohan Mukherjee,1 Nagaraja Rao Padaki,2
Balakumar Reddy,1 Rajesh Gupta,2
Nageshwar Reddy Duvvuru2
1Asian Healthcare Foundation, Hyderabad, India, 2Asian Institute ofGastroenterology, Hyderabad, India
Background: Toll-like receptors (TLRs) play a central rolein sensing and initiating innate antiviral response. Virusesinteracting with host cells can modulate expression andfunction of TLRs for their survival.Objective: To study the expression of TLR2 and TLR4mRNAs in peripheral blood mononuclear cells (PBMCs)of healthy and hepatitis B virus (HBV) infected subjectsand their role in the pathogenesis of acute and variousphases of chronic infection.Methods: Thirty HBsAg positive patients grouped in toacute (AHB, n=9), inactive carriers (IC, n=11), chronic(CHB, n=4), liver cirrhosis (LC, n=3) and hepatocellularcarcinoma (HCC, n=3) were evaluated for serum HBeAg,anti HBe and ALT status by ELISA and biochemical proce-dures. HBV DNA was measured by a real time TaqManPCR assay. TLR 2 And 4 mRNA expression was assessedby reverse transcription PCR (RT-PCR) assay and densi-tometry. Thirty voluntary blood donors served as controls.Results: Both TLR2 and TLR4 mRNAs were significantlyreduced in patients (0.41�0.18; P =0.0002) and (0.46�0.21;P =0.05) in comparison to controls (0.69�0.30 and0.55�0.16).TLR2 was significantly suppressed in AHB(P =0.008) while the ICs showed significant reduction(P =0.01) of TLR4. Expression of both the mRNAs were in-dependent of the corresponding viral load of patients.Conclusion: Suppression of both TLR2 and TLR 4 expres-sion reflect inadequate innate immune mechanism in HBV
nd Experimental Hepatology | March 2013 | Vol. 3 | No. 1S | S8–S14
JOURNAL OF CLINICAL AND EXPERIMENTAL HEPATOLOGY
Molecu
larand
Cellular
Biology
infection. Differential suppression of TLR 2 and TLR 4 inAHB and IC group respectively suggests involvement ofdifferent signaling pathways at different phases of HBV in-fection which may partially explain the mechanism of HBVinduced immuno-tolerance.
Corresponding author. Rathindra Mohan MukherjeeE-mail: [email protected]
PHENOTYPIC CHARACTERISTICS OF FETALLIVER STEM CELLS FROM 6 TO 20 WEEKS OFGESTATION
Padma Priya, Anand Baskaran
EmProCell Clinical Research Pvt. Ltd., Mumbai, India
Background: Fetal liver stem cells are not homogenousbut represented mostly by polyploidy hepatocytes and hep-atoblasts and likewise early hematopoietic cells. Both cellslately used in regenerative medicine. However, fact remainsunclear that, in which period of gestation preferably pro-duces liver parenchyma stem cells and hematopoieticstem cells. In this case practically there is absence of pheno-typic characteristics of liver cells in the dynamics of its de-velopment. Phenotype of cells reflects the status of itsgenome, therefore such information are necessary for fur-ther development of cell transplantation.Aim: To create phenotypic map of fetal stem cells, isolatedfrom abortive material (fetal liver) obtained by means ofMTP on gestation periods from 6 to 20 weeks.Methods: Flow Cytometry phenotypic characteristics ofhematopoietic fetal stem cells: HLA-A, HLA-B, HLA-DR,CD4, CD8, CD12w, CD19, CD24, CD27, CD31, CD34,CD42, CD45, CD56, CD62E, CD62P, CD69, CD74,CD83, CD90, CD100, CD105, CD117, CD133, CD135,CD141, CD144, CD158, CD163, CD184, CD202a, CD326and hepatoblasts/hepatocytes: AFP, CD326, Albumin,C017-1A antigen, MHC, CD68, CD74, CD135, CD326.Results: In dynamics of fetal development significant het-erogenocity expression in fetal liver stem cells studymarkers were established. In particular period predomi-nance in liver cells that expressed erythropoiesis markers,T- and B- lymphocytes and was established period ofKupffer cells appearance. In later stages of gestation(18-20 weeks) among the cells of fetal liver hepatoblastsand hepatocytes were predominated.Conclusion: Obtained results evidence that optimal pe-riod of isolation of early hematopoietic cells (AC133+) isthe 9 weeks gestation period. The most effective transplan-tation of fetal progenitor cells for treatment of liver paren-chymatous diseases could be with help of fetal cells of 18-20 gestation weeks. During this period huge amount ofhepatoblasts and polyploidy hepatocytes could be easilyisolated from fetal liver tissue.
Journal of Clinical and Experimental Hepatology | March 2013 | Vol. 3 | No
Corresponding author. Padma PriyaE-mail: [email protected]
EFFECT OF N-ACETYLCYSTEIN (NAC) INEXPERIMENTAL BLUNT LIVER INJURY
Hayri Erkol, Erdal Yilmaz, Nurettin Kahramansoy,Aysel K€ukner, T€ulin Firat, Mehmet Tosun
Abant Izzet Baysal University, Medical Faculty, Department of GeneralSurgery, Bolu, Turkey
Background and Aims: Blunt liver injury is increasinglypreferred to bemanaged non-operatively. Therefore, agentsthat would increase wound healing are widely investigated.N-acetylcystein (NAC) is an amino acid derivate and a glu-tathione precursor, also an antioxidant. The aim of thisstudy is to investigate the effect of intraperitoneal and in-tramuscular NAC on the bluntly-injured liver.Methods: Intramuscular NAC (imNAC) and intraperito-neal NAC (ipNAC) administered groups were formed, aswell as control groups. These groups were separated forday-3 (imNAC3, ipNAC3, and control3) and day-7 (im-NAC7, ipNAC7, and control7) terminations. Totally, sixgroups were constructed and blunt liver injury was per-formed by the freefall of 200 grams of weight. The dosageof NAC was 50mg/kg and 200mg/kg for intramuscularand intraperitoneal pathways, respectively. AST, ALT,and LDH levels were measured in the serum. Histopatho-logical assessment was done by scoring the inflammationseverity between 0-3 and the Ki67 proliferation index (PI).Results: AST and ALT levels were lower in the day-7groups, compared to those in the day-3 groups. This de-crease was significant in control7 and imNAC7 groups (P#0.02). Among the same day groups, AST, ALT andLDH levels were significantly lower in imNAC and ipNACgroups, when compared with those in the control groups(P =0.01 - 0.004). However, imNAC and ipNAC groupsdidn't differ from each other indicatively (P >0.1). The in-flammation severity didn't differ among day-3 and day-7groups. But, it was significantly low in imNAC3 and ip-NAC3 groups, compared to the control3 group (P=0.010, P =0.049, respectively). Day-7 groups also demon-strated difference. The Ki67 PI's of the imNAC3 and ip-NAC3 groups were prominently higher than those of theimNAC7 and ipNAC7 groups (P <0.001, P =0.005, respec-tively). The Ki67 PI's of the control3, imNAC3 and ipNAC3groups were 1.5, 3.8, 2.1 respectively and the differences be-tween these groups were distinctive (P = 0.001 - 0.038).Discussion: The NAC administration decreases the AST,ALT levels and the inflammation severity; increases cellularproliferation. However, further studies are required.
Corresponding author. Hayri ErkolE-mail: [email protected]
. 1S | S8–S14 S9
