| Background:The skin is the largest organ of protection,however,skin defects caused by various reasons are common in modern society.Because of its advantages of simple and flexible,random flap is usually used to repair the tissue defects caused by trauma,congenital malformation or tumor,It is also one of the most commonly used methods to repair skin defects in clinic.With the rapid development of modern medicine,the majority of scholars are also committed to the production of a variety of animal models to study the skin flap,but also found that many factors affecting the survival of skin flap after transplantation.However,the researchers found that ischemia reperfusion injury is a very common and important factor in the survival of the flap.Due to the lack of well-known vascular flap,the flap is prone to ischemia or reperfusion injury caused by partial or total necrosis of the flap,affecting the healing effect.Ischemia reperfusion injury is the first time that Jennings refers to the phenomenon that the blood supply is restored and the organ injury is aggravated[1].Ischemia reperfusion injury plays an important role in the survival of the flap.In recent years,scholars have done a lot of basic and clinical trials to study the pathological mechanism of ischemia reperfusion injury,however,the specific mechanism is not clear,but it is found that there are free radical damage theory,inflammatory cytokines theory,chemical damage theory and so on.Thus it can be seen,the key to survival of the flap after transplantation surgery is whether to avoid the occurrence of ischemia-reperfusion injury.Therefore,inhibition of inflammatory response is one of the important methods to prevent ischemia reperfusion injury and increase the survival rate of random flap[2-4].Over the years,people have been studying ways to control the inflammatory response,the inflammatory reaction of the body,the highest activity of inflammatory cytokines IL-6,IL-8,TNF-alpha has become the main test indicators in the experiment.Although the use of antibiotics in a wide range of clinical,but the control of inflammation is still not ideal,and the formation of multiple drug resistance and other risks.Therefore,it is a hot spot to find a better anti-inflammatory drug.The researchers found by experiment,Carbachol is a kind of cholinergic receptor agonist,mainly through the N like cholinergic receptors play anti-inflammatory effect.Carbachol is a synthetic cholinergic drugs,its role is very similar to ACH,but its stability is better than ACH,not easy to be hydrolyzed by cholinesterase,so its action time is more durable than rational ACH.Carbachol is a cholinergic agonist,can also activate muscarinic(M)receptors and nicotinic receptor(N),but in recent years a large number of experiments in vitro and in vivo,Carbachol excited choline N like receptors and anti inflammation is closely related to the main mechanism is the activation of N like receptor significantly inhibited TNF-alpha,IL-I beta,IL-6,high mobility group protein B1(high-mobility group box 1,HMGB1)excessive production and release of inflammatory mediators[5].In the past,most of the experiments were conducted to investigate the protective effect of Carbachol on the ischemia reperfusion injury of various important organs in the animal model of burn and scald,At present,the application and research of Carbachol in the treatment of skin flap transplantation is relatively small.In this experiment,the rat model was used to observe the effect of Carbachol on the survival of random skin flap in rats.Purposes:Carbachol has anti-inflammatory effect,which can slow down the ischemia reperfusion injury to a great extent,Promoting the flap survive.The aim of this study was to establish the animal model of rat tail free flap,to explore the protective effect of Carbachol on the survival of random skin flap by means of survival condition,histological observation and measurement of blood inflammatory factors,by fedding with Carbachol.Method: In this study,36 SD rats were divided into A(Carbachol)and B(physiological saline)groups,with each group of 18 rats.First,designing the pedicle random flap model in the back of the rats in the tail end of the pedicle width 2*4cm.Carbachol group in the postoperative administration of choline 30ug/100 g,physiological saline group were given the same amount,once a day,seven days in a row.After seventh days,to observe the survival of the flap,detecte the IL-6,IL-8,and TNFby ELISA after taking out the celiac artery blood,observe the histological changes by HE staining of the distal,middle and proximal skin flaps.All the data were expressed as mean ± standard deviation(?x ±S),and the ttest was used to analyze the difference between two groups.Result:(1)The results of the naked eye:The flap survival was observed on the seventh day after operation:A group:The skin flap showed the growth of hair,good color and good elasticity,and there were few crusts in the distal part of the flap.The flap was lifted,and the granulation and blood vessel growth of the flap were good,and the bleeding was more,and there was no obvious effusion and blood accumulation.B group:The flap of proximal visible hair growth,reddish,elastic;distal skin flap without hair to grow,dark color,inelastic.A small amount of granulation tissue,inflammatory cells and less bleeding were found.(2)Histological observation:A group:A few inflammatory cells could be seen in the distal part of the flap,and there were no obvious inflammatory cells in the middle and the near segments.B group:A large number of inflammatory cells were found in the distal and middle part of the flap,a little in the near segments.According to the macroscopic and microscopic observation of survival of flaps,A group were better than the B group,inflammatory cells of A group were significantly less than group B.(3)Inflammatory factor concentration in serum:The level of IL-6 in experimental group A is(108.14±5.99)pg/ml,group B is(146.21±7.13)pg/ml;The level of IL-8 in experimental group A is(105.12±6.11)pg/ml,group B is(140.41±9.78)pg/ml;The level of TNF-? in experimental group A is(186.75±10.31)pg/ml,group B is(246.27±20.69)pg/ml.The A group IL-6,IL-8 and TNF-alpha expression were significantly lower than B group,and between the two groups of IL-6,IL-8,TNF-alpha expression were significantly different,Carbachol is used for reducing the expression of inflammatory factors IL-6,IL-8 and TNF-alpha were statistically significant(P < 0.01).Conclusion:(1)The injury of inflammatory reaction after flap transplantation showed that IL-6,IL-8 and TNF-were elevated.(2)Carbachol can significantly inhibit the release of IL-6,IL-8 and TNFalpha,reduce the inflammatory response,improve the survival of the rat dorsal random flap,and protect the skin flap. |
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