| Objective: To establish a rat model of Achilles tendinopathy,and to intervene with Zheng’s No.1 new wound medicine to study its control effect,determine the expression of autophagy-related proteins LC3-II and P62 in tendon cells,and explore the influence of autophagy in the Achilles tendon disease.Method:96 male Sprague-Dawley rats were randomly divided into 4 groups: blank control group(C),simple jumping group(J),dressing jumping group(JM),and simple dressing group(M).The time of the materials was randomly divided into 4,6 and 8 weeks groups,8 in each group.Group J was modeled by “electric stimulation jump method”,JM group was modeled by “electric stimulation jump method”,and M group was externally applied with Zheng’s No.1 new wound medicine from the first day of 4 weeks;The general condition and the number of effective jumps were taken.The Achilles tendon tissues were taken at the 4th,6th,and 8th weekends.The histomorphological changes were observed after HE staining.The structures of autophagy in Achilles tendon tissues were observed under transmission electron microscope,and Western blotting was performed.The method was used to detect the expression of autophagy-related proteins LC3-II and P62 in Achilles tendon tissues.The obtained data were statistically analyzed by independent sample T test and Two-factor ANOVA.result:1.Number of jumps: The number of effective jumps in the J group and the JM group showed a downward trend;in the 6th to the 8th week,the number of jumps in the J group was less than that in the JM group,and there was a significant difference at 6 weeks(P<0.05).2.Histomorphology: In group J,the collagen fibers were arranged irregularly at 4 weeks,the gap was slightly increased,a few sputum fibers were interrupted,and the number of nucleus increased.At 6 weeks,the collagen fibers were arranged disorderly and the structure was loose.Breaking,the number of nuclei increased,occasionally inflammatory cells and new blood vessels grow in;at 8 weeks,the collagen fibers are arranged disorderly,sparse,loosely structured,and there is a large area of sputum fiber rupture,a large gap,a large number of inflammatory cells Infiltration,new blood vessels grow in.In the JM group,the collagen fibers were roughly parallel and relatively regular at 4 weeks,and the arrangement was relatively compact.At 6 weeks,the collagen fibers were irregularly arranged,the gap was slightly increased,and a few sputum fibers were broken;at 8 weeks,it was visible.The local achilles tendon collagen fibers are arranged in an irregular manner,a few sputum fibers are broken,and a small amount of inflammatory cells.The collagen fibers of the A group and the M group of the A group were roughly parallel and relatively regular,and the arrangement was compact,the gap was small,and the morphology of the chondrocytes was normal.No significant changes were observed at the 4th,6th and 8th week.Electron microscopy observation: At 6 weeks,autophagosomes were observed in the Achilles tendon tissues of the four groups,and there were fewer autophagosomes in the C and J groups.The number of autophagosomes in the M and JM groups was relatively high.4.LC3-II protein expression: The expression of rats in group J decreased at 4,6,and 8 weeks,and there was no significant difference.The expression in JM group showed an upward trend,and there was significant difference between 4 and 8 weeks.Group C There was no significant difference between the M group and the M group at 4,6,and 8 weeks.At 6 weeks,J group had the lowest,JM group had the highest expression,J group had significant difference with JM group and C group;at 8 weeks,J group had the lowest and JM group had the highest,and J group and JM group had significant difference.5.P62 protein expression: The expression of rats in group J increased at 4,6,and 8 weeks,and there was significant difference between 4 and 8 weeks.The JM group tended to be stable at 4,6,and 8 weeks.No significant difference was observed.The expression of group C and group M showed a downward trend,but no significant difference was observed.At 8 weeks,J group protein expression was highest,and there was a significant difference between group J and group C.conclusion:1.Zheng’s No.1 new wound medicine external application has a certain preventive effect on rat Achilles tendinopathy.2.Insufficient autophagy activity induced by decreased LC3-II and elevated P62 in the Achilles tendon of rats with rickets may be involved in the development of tendinopathy.3.Zheng’s No.1 new wound medicine may improve the effect of tendon disease by increasing the autophagy activity of tendon cells. |
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