Hirokazu Kadoya

Expression of SPARC by activated hepatic stellate cells and its correlation with the stages of fibrogenesis in human chronic hepatitis.

Abstract. Secreted protein, acidic and rich in cysteine (SPARC), which functions in tissue remodeling, has been reported to be expressed by myofibroblasts in liver cirrhosis and hepatocellular carcinoma. This study aimed to reveal its expression in chronic hepatitis. Immuno-light and electron microscopy demonstrated that SPARC was expressed by nerve fibers and hepatic stellate cells (HSCs) in the liver parenchyma and myofibroblasts in the fibrous septa. Reaction products were localized in the rough endoplasmic reticulum and nuclear envelope. Serial section analysis demonstrated that SPARC, platelet-derived growth factor receptor-beta, and alpha-smooth muscle actin were co-expressed by HSCs. Quantitative analysis demonstrated that, while SPARC-positive HSCs were sparse in control livers, they significantly increased in number in the livers with chronic hepatitis. There were, however, no significant differences in number among the grades of activity, the stages of fibrosis, or etiology (virus-infected or autoimmune, hepatitis B virus or hepatitis C virus). In liver cirrhosis, however, they significantly decreased in number. The present results indicate that SPARC is expressed by activated HSCs in chronic hepatitis, suggesting the involvement of SPARC in hepatic fibrogenesis after chronic injuries.

Expression of Cellular Prion Protein in Activated Hepatic Stellate Cells

Suppression subtractive hybridization was used to clone genes associated with the activation of hepatic stellate cells and 13 genes were found to be dominantly expressed in activated stellate cells. Among them, one was identical to the 421-837th base pairs of cDNA sequence reported for rat prion-related protein (PrP). In cultured stellate cells, PrP mRNA expression increased in a time-dependent manner in parallel with smooth muscle (SM) alpha-actin mRNA expression. In situ hybridization demonstrated that PrP mRNA was localized in and around the fibrous septa of carbon tetrachloride (CCl4)-treated liver. Cellular PrP (PrPc) was produced by culture-activated stellate cells, and immunohistochemically detected in the fibrous septa of CCl4-damaged liver and sinusoidal linings of common bile duct-ligated liver, consistent with the localization of SM alpha-actin. Immunoelectron microscopy revealed that PrPc resided on the plasma membrane of stellate cells. These results indicate that PrP expression is closely related to stellate cell activation associated with fibrogenic stimuli.

Clinicopathological characterization of prion: a novel marker of activated human hepatic stellate cells

BACKGROUND/AIMS We recently demonstrated prion as a new marker for hepatic stellate cell activation in rats. Here, we have examined prion expression in normal and diseased human livers. METHODS Prion expression was examined at protein level by immunohistochemistry and at mRNA level by in situ hybridization. RESULTS While normal livers were negative for prion, all liver specimens but one from patients with chronic liver disease were positively stained. In chronic hepatitis, prion protein expression was found not only in the sinusoidal lining cells within the lobules but also in mesenchymal cells in expanded portal tracts. In alcoholic liver disease, prion-positive cells were found mainly in the areas of alcoholic hepatitis. Immunoelectronmicroscopy revealed that prion-positive cells were activated stellate cells. In situ hybridization demonstrated that the distribution of prion mRNA is similar to that of prion protein. In chronic hepatitis, the number of prion-positive cells correlated with the grade of activity but not with the stage of fibrosis. In alcoholic liver disease, levels of prion protein expression were significantly increased in the presence of alcoholic hepatitis. CONCLUSION Prion as a novel marker of activated stellate cells correlates well with disease activity in human chronic liver diseases.

Expression of Fas and Bcl-2 proteins and induction of apoptosis in human hepatocellular carcinoma cell lines

Because the induction of apoptosis in cancer cells is very important in clinical management, it is useful to examine the association with the Fas–Fas ligand pathway and Bcl-2 protein family in apoptosis. We morphologically examined the expression of Fas and Bcl-2 proteins and induction of apoptosis by anti-Fas in four human hepatocellular carcinoma cell lines, PLC/PRF/5, Huh-6, and Huh-7, as well as OCUH-16, which was originally established in our university. Fas protein was expressed in 96% of OCUH-16 cells in cytoplasm, 24% of PLC/PRF/5 cells, 20% of Huh-6 cells, and no Huh-7 cells. Bcl-2 protein was expressed in 43%–72% of cells in cytoplasm and nuclei of the four lines examined. Administration of anti-Fas induced apoptosis in about 40% of OCUH-16 cells, but did not induce apoptosis in the other three cell lines. In conclusion, an original cell line, OCUH-16 cells, expressed Fas and Bcl-2 proteins and underwent apoptosis following treatment with anti-Fas, but the other three cell lines examined did not undergo apoptosis. OCUH-16 cells are thus very useful for the study of apoptosis and molecules related to apoptosis at the levels of cell-surface receptors and intracytoplasmic regulation of apoptosis.

LAPAROSCOPIC WEDGE BIOPSY OF THE LIVER WITH USE OF AN ULTRASONICALLY ACTIVATED SCALPEL

Background: Wedge biopsy of the liver yields more accurate diagnosis of liver diseases than does needle biopsy, but is usually more invasive. We have designed a new method for laparoscopic wedge biopsy under local anesthesia with use of an ultrasonically activated scalpel.

TRENDS IN THE CAUSES AND OUTCOME OF LIVER CIRRHOSIS DIAGNOSED BY LAPAROSCOPY IN OSAKA, JAPAN DURING THE PAST THREE DECADES

Background: Although various factors are assumed to have substantially altered the demographic characteristics of liver cirrhosis, few studies of patients with endoscopically confirmed cirrhosis have been done in Japan. We analyzed trends in causes and outcome of liver cirrhosis in Osaka, Japan during the past three decades.

A 56 Year-Old Female with Congenital Hepatic Fibrosis Diagnosed by Laparoscopy

Abstract: We describe a 56‐year‐old woman with congenital hepatic fibrosis. Blood tests and liver scanning with Tc‐99m‐labelled galactosyl human serum albumin revealed mild liver dysfunction. Per‐rectal portal scintigraphy with iodine‐123 iodoamphetamine showed severe abnormalities in the portal circulation, and the portal pressure measured during percutaneous transhepatic portography was high (350 mmH2O). Idiopathic portal hypertension was suspected. Laparoscopy disclosed diffuse, intense dendritic white markings around the liver. Congenital hepatic fibrosis was confirmed on histologic examination of a biopsy specimen obtained during laparoscopy. In summary, we report a rare and relatively elderly case of CHF, in which laparoscopy was useful in the diagnosis. (Dig Endosc 1999; 11: 174–178)

7006 Laparoscopic wedge biopsy of the liver with use of laparoscopic coagulating shears under local anesthesia.

Wedge biopsy of the liver is more useful compared to needle biopsy for the diagnosis of the liver diseases, especially for the diagnosis of primary biliary cirrhosis or primary sclerosing cholangitis. Wedge biopsy has been done under open laparotomy or laparoscopy with general anesthesia. This method was more invasive than needle biopsy. We performed wedge biopsy laparoscopically under local anesthesia with use of laparoscopic coagulating shears (LCS; Johnson & Johnson Medical, K. K., Tokyo, Japan) from May 1998. LCS is ultrasonically activated scalpel and is a high-frequency oscillating instrument that has both coagulating and dissecting activities. Five patients underwent this wedge biopsy from May 1998 to November 1999. In the five patients, three had autoimmune hepatitis, one had primary biliary cirrhosis, and one had non-alcoholic steatohepatitis. Following the pneumoperitoneum with room air, a 10-mm trocar was punctured at upper right side of umbilicus and a peritoneoscopy was inserted through the trocar. After observation of the surface of the liver as usual, a 10-mm trocar was punctured at upper left side of umbilicus under laparoscopic observation. A LCS were inserted through this trocar. The edge of the liver at subsegment 3 or 4 was dissected with the LCS under output of level 3 as described by the company. Subsequently, the peritoneoscopy and the LCS were pulled out through the trocars. And the peritoneoscopy was inserted through left-side trocar and the LCS was through right-side trocar. The opposite side of the dissected edge was resected with LCS. During resection of the liver, the patients complained slight abdominal pain but no troubles were observed. The wedge biopsied specimens were pulled out with a 5-mm forceps through the trocar. The bleeding was few and no serious complications were found. Tissues were placed in formalin for histological examination. Microscopic findings showed mild injury along the dissected portion but the central area of the specimens were intact. So no troubles for the diagnosis of the tissues were experienced. Compared to needle biopsy, more sufficient amount of tissues were obtained with this method. After laparoscopic wedge biopsy, all 5 patients complained mild abdominal pain and slight fever. But these symptoms were same degree as of the patients after laparoscopic needle biopsy. Conclusions: Laparoscopic wedge biopsy with LCS under local anesthesia was safe and more useful compared to needle biopsy. And this method was no more invasive than needle biopsy.

7004 Trends in the causes and outcome of liver cirrhosis diagnosed by laparoscopy in japan during the past 3 decades.

Background: Although various factors are assumed to have substantially altered the demographic characteristics of liver cirrhosis, few studies of patients with endoscopically confirmed cirrhosis have been done in Japan. We analyzed trends in causes and outcome of liver cirrhosis in Japan during the past 3 decades. Methods: The study group comprised 466 patients with cirrhosis of the liver who underwent laparoscopy at any time between 1969 and 1998. To analyze trends in the causes and outcome of cirrhosis, the study period was divided into three 10-year intervals, variables included hepatitis B surface antigen, hepatitis C virus antibody, alcohol intake, causes of death, and outcome. Results: HCV has remained the major course of liver cirrhosis during the past 30 years. Before the discovery of HCV in 1989 most cases diagnosed as non-A, non-B hepatitis were caused by HCV infection. Alcohol played a part in the development of liver cirrhosis in more than 30% of the study group, alone or with hepatitis viruses, alcohol therefore remains an important factor in the pathogenesis of cirrhosis. The outcome of cirrhosis has improved gradually during the past decade owing to more effective means of treatment for complications of cirrhosis, such as esophageal varices, HCC, and hepatic encephalopathy. Conclusion: HCV has remained the major cause for the past 3 decades. Improvement of outcome in patients with liver disease caused by HCV infection is an important task.