Immunohistochemical staining were done for Ki67 (hepatocyte replication) & CK19 (HPC & intermediate hepatocytes) for characterization of nature of hepatic regeneration & CD68 (liver tissue macrophages) CD163 (M2 macrophages)
for analysis of macrophages in liver tissue samples from patients with ACLF (n=15), ALF (n=21), CLD, (n=22). Expression CD68 & CD163 macrophages were correlated with hepatocyte replication, HPC activation & maturation. Results RT-PCR analysis documented significant increase in M2 gene markers CD163 CD206 & TGM2 (p=.001, .001 & .002) & decrease in M1 markers iNOS & CD80 (p=.001, .001) in ACLF comparison to CLD. Similarly there was significant
increase in M2 gene markers CD163, CD206 & TGM2 (p=.002, .002 & .002) & decrease in M1 markers iNOS CD80 (p=.002, p=.002) in ACLF comparison to ALF. Immunohistochemical analysis shows increase in Ki67+ hepatocytes GSK3235025 in ALF as compared to ACLF & CLD (p=.0001, .0001). Further, the number of CK19+ HPC & its maturational lineages was increased in ACLF than ALF & CLD (p=.0001, .0002). Using spearman rho correlation shows that CD163 positivity & M2/M1 macrophage ratio is significantly associated with extant of HPC differentiation to hepatocyte (p=.0001, .0004). Further Pu1 (yolk sac originated Kupffer Cells) & Myb (bone marrow originated monocyte derived macrophage) expression suggest significant increase in Pu1 expression relative to CD68 in ACLF in comparison to ALF & CLD (p=0.002, 0.001) suggesting majority of M2 macrophage in ACLF are kupffer JAK inhibitor either cells.
Conclusions Alternatively activated M2 macrophages are major population in ACLF liver which promotes differentiation of HPC to hepatocyte. These M2 macrophages are of kupffer cell origin. Disclosures: The following people have nothing to disclose: Dhananjay Kumar, Sheetalnath Rooge, Smriti Shubham, Adil Bhat, Charvi Syal, Archana Rastogi, Chhagan Bihari, Viniyendra Pamecha, Anupam Kumar, Shiv K. Sarin Background/aim: Patients with HCV/HIV co-infection show faster progression of liver fibrosis, in part associated with miRNA dysregulation and more severe inflammation. The HIV protein gp120 modulates directional migration and expression of pro-fibrogenic cytokines in hepatic stellate cells (HSC), through engagement of the chemokine receptor CCR5. The NALP3 inflammasome is a critical pathway in the generation of pro-inflammatory signals during liver injury. Aim of this study was to evaluate the role miRNAs and inflammasome activation in mediating the effect HSC. Methods: HSC were isolated from normal human liver tissue. Inflammasome complex gene expression was measured by qRT-PCR. Levels of mature IL-1 β were assayed by ELISA. miRNA expression was evaluated via RT-PCR.