Study On The Apoptosis Induction And Mechanism Of Hirudin On Human Hypertrophic Scar Fibroblasts

Hypertrophic scars represent an aberration in the fundamental processes of wound healing. It is one of the most common pathological phenomenon seen after wound healing of trauma, especially in deep burned patients. Abnormal Proliferation of fibroblast and excessive deposition of extracellular matrix responsible for raised. Therefore, it is significant that exploring the mechanism of HS formation to look for effective prevention and treatment. Many Published reports documented the mechanism and advoeated avariety of therapies, however, few studies Provide a coherent therapeutic Plan. So to seek the effective way and medicine for prevention and cure HS is a very important subjection on plastic surgery science. Hirudin has various biological effects in smoothing muscle cells proliferation, influencing the inflammatory cytokines secretion of monocyte. Thus, we design the experiment to observe effect of hirudin On HS to exploring the effective treatment method and mechanism of HS, provide the theoretical foundation of clinical medicine.ObjectiveTo study the apoptosis induction and mechanism of hirudin on human hyperplastic scar fibroblasts in vitro.MethodsHS fibroblasts were isolated and cultured by routine method.The 4一7 generation cells could be used. The final conecentrations of hirudin for experimental group were: 1u/ mL,0.5 u/ mL,0.25 u/ mL,0.125 u/mL. Detecting indexes:1 Observed cells` appearance change by inverted phase contrast microscope;2 Detected the cells` vigor and proliferation by MTT method;3 Evaluating the expression of bax,bcl-2 in human hypertroic scarfibroblasts were cultured in vitro after 24 hours .4 The expression of bax and bcl-2 were detected by Western blot.Results1 The result of MTT showed that the proliferative tendency of experimental HSFb presented inhibited correlating with does of hirudin;2 The percentage of apoptosis was increased in dose of hirudin depentent way, compare with the control group , It has the statistical significance;3 Hirudin could increase the bax and inhibit the bcl-2 expression of HSFb;ConclusionThe experiment proved hirudin has a distinguished inhibitory action to HSFb by detect the different index of HSFb culture in vitro. Those mechanism of action may be relate to below several aspects:1. The hirudin can cause the alteration in cell morphology of HSFb;2. The hirudin to inhibit the proliferation of fibroblasts;3. The hirudin can increase obviously the expression of bax of HSFb;4. The hirudin can inhibit obviously the expression of bcl-2 of HSFb.