| Objective:This experiment aims to observe the effect of external Chinese medicine ointment developed under the guidance of "simmer pus and grow meat" method on the expression of aFGF,Caspase-8 in the new granulation tissue in the process of wound repair in rats with postoperative anal fistula trauma model through molecular biology and morphology from the perspective of cytokines and apoptotic protease regulating cell activity,analyze the intervention effect and possible mechanism of Wugu Kirin Ointment on acute,open and infectious wound,and provide reference for further study of cell signal transduction and related signal pathways involved in wound healing process of anal fistula,at the same time further scientifically expound and prove traditional Chinese medicine surgery classic theory "simmer pus and grow meat" method and expand its scope of application,to promote the postoperative anal fistula wound healing to develop effective Chinese medicine preparation which can be put into clinical application,to pave the way for clinical application of Wugu Kirin Ointment.Method:120 experimental rats were randomly divided into blank group,Medical White Vaseline Ointment group(model group),Jingwanhong Ointment group(control group)and low,medium and high dose Wugu Kirin Ointment group(experimental group).In addition to the blank group,the whole skin excision injury was made in the back of the rats in the other groups,and the infectious open wound model was established by sprinkling fecal suspension.After the successful modeling,each group was given drug intervention,respectively draw the new granulation tissue to fix and set aside on the 3rd,7th,11 th and15th days after administration.General conditions,wound healing time and wound healing rate were observed and recorded with naked eye.By means of hematoxylin-eosin staining(HE staining),the morphological and structural changes of the new granulation tissue in each group were observed under microscope,and the expression difference of aFGF,Caspase-8 in the new granulation tissue of each group was detected by immunohistochemical staining(IHC staining),and the number of cell apoptosis in the new granulation tissue of each group was detected by TdT-mediated dUTP Nick-End Labeling(Tunel).Results:1.After the drug intervention,the rats in the control group and experimental group were superior to the model group in eating,drinking water,activity and wound condition,as well as the morphology and structure of the new granulation tissue in the wound under light microscope.The general condition and morphological changes of granulation tissue in the low-dose experimental group were similar to those in the control group,while the medium and high-dose experimental groups recovered faster than the control group,and the vital signs,wound condition and morphological characteristics of the granulation tissue under light microscope in the high-dose experimental group were better.2.Compared with the model group,the wound healing time of the control group and the experimental group was shortened(P<0.05),and the wound healing rate of the rats on the 7th,11 th and 15 th days after administration was increased(P<0.05).There was no significant difference in wound healing time and wound healing rate between the low-dose experimental group and the control group(P>0.05),while the wound healing time of the medium and high-dose experimental group was shorter than the control group(P<0.05,P<0.01),and the wound healing rate of the medium and high-dose experimental group on the 7th,11 th and 15 th days after administration was higher than the control group(P<0.05,P<0.01).3.Compared with the model group,the aFGF content in the new granulation tissue of the control group and the experimental group increased on the 3rd,7th and 11 th days after administration(P<0.05),and the aFGF content decreased on the 15 th day after administration(P<0.05).There was no significant difference in the aFGF content of the new granulation tissue between the low-dose experimental group and the control group at each time point(P>0.05),while the aFGF content of the new granulation tissue in the medium and high-dose experimental group was higher than the control group on the3 rd,7th and 11 th days after administration(P<0.05,P<0.01),and the aFGF content of the new granulation tissue in the medium and high-dose experimental group was lower than the control group on the 15 th day after administration(P<0.05,P<0.01).4.Compared with the model group,the expression level of Caspase-8 and the number of cell apoptosis in the new granulation tissue of the wound in the control group and the experimental group decreased(P<0.05).There was no significant difference in the expression level of Caspase-8 and the number of cell apoptosis in the new granulation tissue of the wound between the low-dose experimental group and the control group(P>0.05),while the expression level of Caspase-8 and the number of cell apoptosis in thenew granulation tissue of the wound in the medium and high-dose experimental group was lower than the control group(P<0.05,P<0.01).Conclusion:1.Wugu Kirin Ointment can obviously promote the wound in rats with postoperative anal fistula trauma model healing,especially the effect of high-dose Wugu Kirin Ointment is better.2.The possible mechanism of Wugu Kirin Ointment to promote the wound healing is to improve the growth rate and quality of the granulation tissue to promote the wound healing by regulating the expression of aFGF,Caspase-8 to change the number of multiple cell proliferation and apoptosis in different healing periods. |
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