Expression And Significance Of TRAIL And DR5 And NF-κB In Human Chronic Granulation Tissue And Hypertrophic Scar

BackgroundThe scar is the inevitable result of a variety of repair wounds to heal.It took place in the main trauma,infection,surgery and burn wounds,and so on,a wide range. Pathological scar in hypertrophic scars is based on fibroblast proliferation and excessive extracellular matrix for the excessive accumulation of the main characteristics of clinical hypertrophic scars only within the boundaries of the original wound growth.The Pathogenesis is not quite so far.The clinical performance for the local pain,itching,and hyperplasia with varying degrees of dysfunction and endanger people's physical and mental health.In the prevention and treatment,there is no effect on the methods of plastic surgery research.It is a hot and difficult problems.Apoptosis and cell proliferation are essential to life,the number of cells in the body is to maintain the dynamic equilibrium of the basic measures.In recent years, apoptosis in apoptosis factor Tumor necrosis factor(TNF)-related apoptosis-inducing ligand(TRAIL) and its receptor become a hot research.TRAIL widely found in various tissues and organs,can lead to abnormal directed to apoptosis,and will not affect normal cell growth and differentiation.TRAIL-induced apoptosis mainly get through the cytoplasmic or extracellular pathway activated by.In hypertrophic scars, the TRAIL death receptor 5(DR5) plays an important role.More and more studies show that many of the diseases that are related to the development of TRAIL apoptosis,in order to TRAIL as a target for treatment or mitigation of disease has become a hot research and clinical trials have been applied in.Nuclear factor-kappaB(NF-κB) signaling pathway get through the healing of wounds and repair the entire biological process.NF-κB is a more forward role in the regulation of transcription factors,many of the cells of gene expression plays a crucial role in regulation.It is also involved in cell growth,development,apoptosis and other physiological functions,and malignant cell transformation,such as fibrosis plays an important role.More and more studies have shown that the incidence of many diseases and the development and the over-activation of NF-κB.The NF-κB in the expression of p65 to prevent apoptosis and to improve the treatment of TRAIL cells in the activity of NF-κB.The comparison of TRAIL induced apoptosis signaling and other tumor necrosis factor(TNF) family members is not clear,especially for TRAIL how to control activity of NF-κB and how to overcome the anti- apoptosis role of NF-κB.How to regulate the activity of NF-κB in TRAIL and how to overcome the anti- apoptosis role of NF-κB has not been reported at home and abroad.It study different stages of hypertrophic scars and granulation tissue for the study to control for normal skin,using immunohistochemistry,RT-PCR and Western blot detection of TRAIL and its death receptor DR5 and NF-κB in various stages of hypertrophic scars.The expression to explore the mechanism of scar for the clinical treatment of hypertrophic scars to provide a new direction and ideas.ObjectiveThis study was to collect the clinical chronic granulation tissue,hypertrophic scars and normal skin samples to observe the TRAIL signaling pathway in TRAIL-induce apoptosis and its receptor DR5 and the p65 gene in NF-κB signaling pathway key organizations in different stages the level of signaling expressed by the existence of differences.In the choice of chronic granulation tissue,within six months,six months to one year and more than a year hypertrophic scars,hypertrophic scars observed apoptosis-TRAIL,apoptosis receptor DR5 and NF-κB p65 change in the impact of dynamic changes,and compared with normal skin samples.MethodsThe collection of clinical specimens,including 40 cases of various stages,10 cases each group,of human chronic granulation tissue,hypertrophic scars,and 8 cases of normal skin.Using immunohistochemistry analyse the samples TRAIL,NF-κB p65 protein expression;Real-time quantitative RT-PCR testing organization TRAIL and DR5 and NF-κB p65;Western blot to detect the level of TRAIL and NF-κB p65 protein expression.Experimental group:Group A for the chronic granulation tissue,hypertrophic scars six months for group B,six months to one year for hypertrophic scars for Group C,more than a year hypertrophic scars for the group D,the control group of normal skin group for the group E.Results1.Biopsy HE staining and immunohistochemical staining SP method HE staining showed that the groups are in the corresponding pathological manifestations. Group A can be seen in a large number of endothelial cells and capillaries,as well as many of the new round of fibroblasts and collagen fibers to form a small, inflammatory cell infiltration.Group B to Group D in fibroblasts decreased gradually shape into a round fiat by the slender,inflammatory cells and capillaries gradually decreased to disappear from the thick skin collagen fibers,the disorder gradually thinning.But the rules of order is thiner than the epidermis on the normal squamous. Non-sudden foot and skin appendages.Group E level of epidermal clear,obvious keratosis keratosis layer,skin adnexal rich in collagen fibers arranged in the rules,and less loose,curly,small cells.2.Immunohistochemical staining SP method SP showes TRAIL in group A and B expressed strong positive,strong in the group C,D and E;positive expressiones of group C,D are gradually weakened,close to of group E;NF-κB p65 expression in Group A and Group E.Groups B,C,D expressed positive.In particular,noted that the NF-κB p65 in Group B in basal layer of skin is particularly positive.3.RT-PCR TRAIL level Biopsy HE staining and immunohistochemical staining SP method of activity in group A and group B at a peak,were significantly higher than other groups(p＜0.05),while group A and group B no statistically significant difference(p＞0.05),C group,D group and Group E in each group were compared statistically no significant difference.(P＞0.05).DR5 level of activity in group A peak was higher than that of E group(p＜0.05), hypertrophic scar formation of the various stages of B group,C and D groups to express their level of activity significantly lower than that of A group(p＜0.05).B,C, D groups were compared between the groups no statistically significant difference(p＞0.05).B,C,D groups were lower than the E group(p＜0.05).NF-κB p65 activity levels were compared between the groups was no significant difference(p＞0.05).However,the group look at the number,E Group for 0.30,A Group for 0.38,B Group for 0.44,C group was 0.46,D Section 0.47.Group E and Group A at a low level,B,C,D group at a relatively high level.B,C,D groups are similar,and there is higher than that of Group E and Group A.TRAIL and DR5 in linear correlation analysis,Pearson correlation coefficient 0.291,P=0.045＜0.05,that TRAIL and DR5 positive correlation exists.NF-κB p65 in TRAIL and linear analysis,p65 and TRAIL the Pearson correlation coefficient 0.371,P=0.009＜0.05,and that p65 is related to the existence of TRAIL.In the NF-κB p65 and DR5 linear correlation analysis,Pearson correlation coefficient -0.088,P=0.554＞0.05,and DR5 that p65 does not exist related to each other.4.Western blot test results TRAIL in Group A and Group B in a higher level of protein expression in the C group,D group,E group close to the level of protein expression,Statistical analysis,A and group B protein expression was significantly higher than other groups(p＜0.05).NF-κB p65 in Group A and Group E protein expression is no significant difference in the level of statistical significance(p＞0.05),B group,C group,D group of protein expression levels were significantly higher than that of group A and E group(p＜0.05). Conclusion1.In the chronic granulation tissue,the levels of apoptosis TRAIL and death receptor DR5 are significantly higher,NF-κB p65 no significant changes in the chronic granulation tissue showed that apoptosis increased,the level of anti-apoptosis is inhibited,the organization difficult to heal.2.In the early hypertrophic scars(in six months),TRAIL expression is still high, but its death receptor DR5 significantly lower than normal,NF-κB p65 expression was significantly increased.Skin significantly enhance the role of anti-apoptosis,and apoptosis decreased significantly the performance of scar tissue hyperplasia role.As the basal cell layer of skin has the ability to split activly,which showed that NF-κB p65 may be in the proliferation of skin plays an important role in apoptosis of the skin has a strong inhibitory effect on skin,and plays an important role in proliferation.3.In the latter of hypertrophic scars(6 months to 1 year ),TRAIL and its death receptor DR5 are at a low level of apoptosis,NF-κB p65 expression was still higher than normal skin,show that in the latter part of hypertrophic scars still hyperplasia.4.In the mature hypertrophic scars(after 1 years),TRAIL and NF-κB p65 expression levels close to normal skin.Scar tissue to strengthen the role of apoptosis, proliferation weakened,HS become flat and faded.5.NF-κB p65 and TRAIL there is a positive correlation,showing that enhancing the level of expression of TRAIL will induced NF-κB p65 activity,while NF-κB p65 gettes through the gene transcription regulation to regulate the expression of TRAIL. TRAIL and DR5 positive correlation exists that the TRAIL receptor DR5 expression changes,to a certain extent,reflected the cells of the sensitivity of TRAIL-induced apoptosis.SignificanceFirst to detect the TRAIL,death receptors DR5 and NF-κB p65 in the chronic granulation tissue and hypertrophic scars in different stages of the proliferation expression,suggesting that the TRAIL apoptosis pathways and NF-κB p65 may be mostly involved in abnormal apoptosis reduction of chronic granulation tissue repair and in the unusual apoptosis of scar hyperplasia.It provides a theoretical basis for the foundation of the hypertrophic scars treatment and applies the timing of the treatment. This is for us in the clinical treatment of hypertrophic scars and scientific research to provide a new direction and ideas.