Effects of FK506 on Bcl-2 mRNA and Bax mRNA expressions as well as cell apoptosis in rat limb allografts / 中国组织工程研究

ABSTRACT

BACKGROUND: In the process of limb allograft, apoptosis of target cell is one of the main mechanisms of dysfunction of allograft, which might lead to the failure of allotransplantation. It is assumed that immunosuppressant may relate with cell apoptosis. OBJECTIVE: To investigate effects of FK506 on Bcl-2 mRNA and Bax mRNA expressions and cell apoptosis in rat limb allograft. DESIGN, TIME AND SETTING: The randomized controlled animal trial was performed at the Animal Experimental Center of First Hospital of Harbin Medical University from June 2005 to November 2006. MATERIALS Fifty-six clean-grade healthy male SD recipient rats and 56 Wistar donor rats were selected. FK506 was product of Fujisawa, Japan (No. 100143G). METHODS: Right hind limb was separated from the upper segment of thigh of SD rat (donor), and washed using heparin saline. The recipient rates underwent limb allotransplantation from allogenetic Wistar to establish injury model. The recipients were randomly divided into two groups (n=28) immunosuppressant group was injected with FK506 1 mg/kg per day, and the control group was not given any immunosuppressant. The right hind limb including skin, subcutaneous tissues, muscles and femoral arteriolar-venular tissue mass were harvested on postoperative days 1, 3, 5, and 7. MAIN OUTCOME MEASURES: Bcl-2 Mrna and Bax mRNA expression were detected using reverse transcription polymerase chain reaction (RT-PCR), and cell apoptosis was detected using in situ terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling technique. RESULTS: Fifty-six model rats were included in final analysis. On the 3, 5, and 7 days after FK506 injection, Bcl-2 mRNA expression in immunosuppressant group was significantly higher than the control group (t=7.18-21.20, P