The Experimental Study On The Effect And Mechanism Of Emodin To Fibroblast In Hypertrophic Scar In Vitro

Hypertrophic scar(HS) is one of the major unsettled clinical problems that due to abnormal proliferation of fibroblast and excessive deposition of extracellular matrix.The scars most commonly occur when epithelialization has been delayed during,foe example, the healing of deep dermal burn wounds.The HS are thick and raised and often darker in color than surrounding skin,moreover,that are frequently associated with pruritus and pain. 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 advocated a variety of therapies;however,few studies provide a coherent therapeutic plan. Applying traditional Chinese physician to occur the HS is the idea treatment method because of lots of advantages,e.g.extensive resources,cheap price,widely clinical application and light adverse effect.Emodin(3-methyl-1,6,8-carboxyl-anthraquinone) is derivate of hydroxyanthraquinone and an effective component isolated from many kinds of herbs.Generous studies prove that emodin has various biological effects in inhibiting tumor cells and smoothing muscle cells proliferation,influencing the infammatory cytokines secretion of monocyte.Thus,we design the experiment to observe effect of emodin on HS to exploring the effective treatment method of HS and provide the theoretical foundation of clinical medicine.Method:HS fibroblasts were isolated and cultured by routine method.The 4-6 generation cells could be used.The final concentrations of emodin for experimental group were:20ug/ml,40 ug/ml and 60 ug/ml.Detecting indexes:1.Detecting the improves of cell proliferation using the method of MTT.2.FACS detecting the changes of cell cycle.3.FACS checked the condition of apoptosis.4.Laser focusing detected the ion concentration of intracellular Ca²⁺ and the liberation. 5.³ H-TdR detected the synthesis of collagen protein.6.RT-PCR to checked the transcription of mRNA ofâ… type collagen.7.Immunohistochemistry detected TGF-β₁8.Immunohistochemistry detectedα-smooth muscle actin.Result:1.The result of MTT showed that the proliferative tendency of experimental HSFb presented inhibited correlating with does of medicine2.By administration of emodin,the detection of cell cycle showed that:the percentage of G0/G1 was increased while the percentage of S phase and G2/M was decreased.The result demonstrated that emodin could block the cells enter into phase S and hindered the DNA synthesis.3.By administration of emodin,the percentage of apoptosis was increased in dose of emodin dependent way.4.By administration of emodin,The intracellular Ca²⁺ of HSFb were released from the calcium library to inhibit HSFb proliferation.5.The decreased collagen synthesis of experimental HSFb with the relationship with the time.6.Emodin could inhibit the transcription of mRNA ofâ… type collagen.7.Emodin could inhibit the TGF-β1 expression of HSFb.8.Emodin could inhibit theα-SMA expression of HSFb.Conclusion:To detect different indexes of HSFb by administration of emodin,we proved that emodin could significantly inhibit HSFb proliferation.The mechanism maybe including: changing the morphous of cells,blocking of DNA synthesize,enhancing release of intracellular Ca²⁺,inhibiting synthesis of collagen protein,promoting HSFb apoptosis and inhibiting the expression of TGF-β1,α-SMA.