Hepatic sonic hedgehog protein expression measured by computer assisted morphometry significantly correlates with features of non-alcoholic steatohepatitis

Abstract

BackgroundHepatic expression of Sonic Hedgehog (SHH) is associated with Non-alcoholic fatty liver disease (NAFLD) and development of Non-alcoholic steatohepatitis (NASH). Hepatic SHH detection increases with the diagnosis of NASH. This pilot study was designed to confirm that staining for SHH is useful in NASH diagnosis and determine whether quantification of staining by computer assisted morphometry (CAM) can be used to assess severity of ballooning degeneration.MethodsSHH was detected by immunohistochemistry (IHC) on paraffin-embedded liver sections in subjects (N = 69) with biopsy proven NAFLD and no liver disease (control). Serum samples were also available for these subjects. Post-staining, a digitized image of the section was acquired and an area quantification algorithm was used to quantify the degree of SHH expression. Additionally, circulating M30, M65, and SHH were measured by ELISA.ResultsNotably, hepatic SHH expression correlated with histologic ballooning degeneration (rho = 0.62, p < 0.0001), steatosis grade (rho = 0.554, P < 0.001), Mallory-Denk bodies (rho = 0.54, P < 0.001), pericellular fibrosis (rho = 0.527, P < 0.001), and lymphocytic infiltration (rho = 0.435, P < 0.0002). Additionally, hepatic SHH expression correlated with circulating M65 (rho = 0.588, p < 0.0001), and circulating M30 (rho = 0.375, p = 0.001), as well as AST and ALT (rho = 0.43, p = 0.0004, and rho = 0.27, p = 0.03, respectively). Further, serum M30 was almost twice as high in NASH patients compared to non-NASH (539.1 ± 290.8 U/L vs. 287.6 ± 190.5 U/L; p = 0.0002), while M65 was almost three times higher in NASH patients compared to non-NASH (441.2 ± 464.2 U/L vs. 162.8 ± 353.1 U/L, P = 0.0006). Logistic modeling indicates hepatic SHH expression and presence of type 2 diabetes as independent predictors of advanced fibrosis (defined as portal and pericellular fibrosis > 2: OR = 1.986, p = 0.01, and OR = 3.280, p = 0.03, respectively).ConclusionThus, our findings show quantitation of SHH expression by CAM can provide a tool for quantifying changes in hepatocyte injury and assist in unambiguous staging/grading of NASH. Our study showed minimal interobserver variability using CAM based quantification. Once validated, CAM assessment of hepatic SHH could benefit clinical trials or long term outcomes studies of NASH subjects.

Steatosis as main determinant of portal hypertension through a restriction of hepatic sinusoidal area in a dietary rat nash model

“…cance. Although the increase in Shh expression goes in line with the increase in NASH histological activity observed in the 36w-HFGFD group,29 it is worth noting that ballooning did not change over time in our model. Thus, it is indeed hard to discern in our model whether this increase in Shh expression was related to subclinical changes (ie not obvious as ballooning at light microscopy) in lipotoxic hepatocytes or to other potential cellular sources (HSC, LSECs, inflammatory cells), or how this might be related or not with ED.Finally, it is worth remarking the changes observed in the 36w-CD group.…”

miR‐134 inhibits osteosarcoma cell invasion and metastasis through targeting MMP1 and MMP3 in vitro and in vivo

“…The SI was scored as 0, negative; 1, weak; 2, moderate; and 3, strong staining, while the PP was scored as 0, negative; 1, more than 0 but less than 10% positive cells; 2, 11–50% positive cells; 3, 51–80% positive cells; and 4, more than 80% positive cells. At least three microscopic fields (×200) per section were evaluated and scored for the IRS according to previous studies .…”

FGF21: An Emerging Therapeutic Target for Non-Alcoholic Steatohepatitis and Related Metabolic Diseases

“…Stressed and injured NASH hepatocytes express and secrete a second Hedgehog ligand, Sonic Hedgehog (SHH), which is positively correlated with hepatocyte ballooning, steatosis, pericellular fibrosis, and fibrosis stage in liver samples from NASH patients (206,207). Hepatocytes that highly express SHH appear colocalized within fibrotic areas in livers of NASH patients (208), and are proximal to areas of hepatic progenitor cells (209), consistent with the observation that SHH signaling promotes and maintains a de-differentiated liver cell state (210).…”