Explore the Therapeutic Effect of Extracorporeal Shock Wave Therapy on Liver Fibrosis

According to the Ministry of Health and Welfare statistics, Taiwan has a population of 21.2 per 100,000 people died of chronic liver disease and liver fibrosis in 2014. Chronic liver disease and hepatic fibrosis ranked ninth in the top ten causes of death in Taiwan. Shock wave is defined as having a high peak pressure (100 MPa), is a rapid increase in pressure (＜10 ns) and short period (10 micros) single sonic pulse sequence. Extracorporeal shock wave (ECSW) generator is suitable to deliver the energy density range of 0.03 ~ 0.15 mJ / mm2 to a specific target area. Extracorporeal shock wave treatment to inhibit inflammation, reduce oxidative stress and anti-apoptotic effects. There has been no report of using ECSW to treat liver fibrosis and promote regeneration of damaged liver cells. In this project we will examine the therapeutic effect of ECSW therapy on liver fibrosis in rats. The experimental animals were divided into three groups, each group of 12 rats to do: (1) normal group, (2) liver fibrosis group, (3) ECSW treatment group. Hepatic fibrosis rats after repeated ECSW therapy five weeks will take to be sacrificed for taking blood and liver tissue to do the following analysis: 1. Blood analysis AST, ALT and albumin. 2. Masson trichrome staining of liver collagen fibers. 3. Image Analysis the percentage of liver fibrosis area. 4. Immunohistochemical staining of liver macrophage marker CD68, macrophages secrete cytokines TGF-, cell apoptotic marker TUNEL, cell proliferation marker PCNA and hepatic stellate cell activation marker α-SMA expression. 5. Immunofluorescence staining of liver SDF-1, CXCR4, mesenchymal stem cell marker CD44, M2 macrophage marker CD206 / F4-80 expression. 6. ELISA analysis of liver HGF, SDF-1, inflammation markers IL-1, IL-6 and TNF-, anti-inflammatory marker IL-10 expression. 7. western blot analysis of liver cells apoptotic markers Bax and caspase 3, anti-apoptotic marker Bcl-2, cell proliferation marker PCNA, oxidative stress markers NOX2 NOX4,anti-oxidative stress markers HO-1 and NQO-1, inflammatory markers IL -1, NF-kB and TNF-, albumin, MMP9, α-SMA, CD68, pAKT, CD44, CD206, SDF-1 and CXCR4 expression. 8. Quantitative real-time polymerase chain reaction analysis of liver type I collagen, TGF-, CD44, CD206 gene expression. Preliminary results show that ECSW treatment of liver fibrosis in rats can significantly decrease AST, significantly increased albumin to near normal level and significant reduction in liver fibrosis area.

Project ID:PB10608-3665
External Project ID:MOST106-2221-E182-048