Cannabinoid Receptor 1 Antisense Oligonucleotide Treat Liver Fibrosis Mice Study

According to the Ministry of Health and Welfare statistics, Taiwan has 21.2 people per 100,000 population died of chronic liver disease and liver fibrosis in 2014, chronic liver disease and liver fibrosis in ranked ninth in the top ten causes of death in Taiwan. Endocannabinoid system includes cannabinoid receptors, endogenous cannabinoid ligands. Cannabinoid receptors have cannabinoid receptor type 1(CB1) and cannabinoid receptor type 2(CB2). The endocannabinoid system is involved in the pathogenesis of liver fibrosis, for example, to obtain a specimen of liver fibrosis patient from surgery is a strong CB1 protein expression, but normal human liver specimen is a relatively weak CB1 protein expression. Antisense oligonucleotide inhibits the expression of the target gene. In this research proposal we will use CB1 antisense oligonucleotides for the treatment of liver fibrosis in rats’ feasibility study. Experimental animals will divide into 5 groups, 12 mice in each group to do: (1) normal mice, (2) liver fibrosis mice, (3) CB1 antisense oligonucleotide treatment group, (4) CB1 sense oligonucleotide treatment group, (5) CB1 antagonist AM251 treatment group. After 28 days of treatment, liver fibrosis mice will be sacrificed to take blood and liver tissue for making the following analysis: 1. Blood biochemical analysis AST, ALT and albumin. 2. Western blot analysis liver tissue albumin, CB1, α-SMA, and MMP9 protein expression. 3. Masson's trichrome analysis percentage of liver fibrosis area. 4. Immunohistochemistry analysis liver tissue CB1, type I collagen, α-SMA, and TGF-β expression. 5. Quantitative real-time polymerase chain reaction analysis liver tissue type I collagen, α-SMA, TGF-β, and CB1 gene expression. 6. ELISA analysis liver tissue IL-6, IL-10, and HGF. Preliminary results show that the use of CB1 antisense oligonucleotides for the treatment of hepatic fibrosis mice significantly reduces the AST, ALT and significantly decrease liver fibrosis area.

Project ID:PB10507-2467
External Project ID:MOST105-2221-E182-071