Distribution And Imaging Difference Of Reactive Oxygen Species In Muscle Tissue Of Rat Abdominal Wall

In the previous study,we used two radical-reactive fluorescent probes,DCFH-DA and Mito-SOX^TM Red on mice,New Zealand rabbits and cynomolgus monkeys,and found that free radicals were mainly distributed in the connective tissue of linea alba.At the same time,the braided ligament of New Zealand rabbits was stained.According to the staining image,it was speculated that reactive oxygen species could be transmitted in a specific way by relying on collagen fibers in connective tissue.In order to study the distribution of reactive oxygen species in the linea alba of SD rats,we first homogenized the easily observed linea alba and compare the rectus abdominis muscle.By measuring the related indicators of oxidative stress,active oxygen free radical content,antioxidant enzyme activity and antioxidant capacity of the two tissues were compared.From the perspective of biochemical indicators,further explored the distribution of reactive oxygen species in linea alba and rectus abdominis muscle.Then,the sections of linea alba and the rectus abdominis muscle were observed by microscopy,such as HE staining,Masson staining and toluidine blue staining.From the perspective of morphology and histology,explored the distribution of reactive oxygen species in linea alba and rectus abdominis muscle.Finally,the phenomenon of the flow of a specific fluorescent probe appearing on the linea alba was observed by a fluorescent probe method.The main findings were as follows:1.On the abdominal wall of SD rats,the content of reactive oxygen species in the linea alba was higher than that in the rectus abdominis muscle,such as H₂O₂?O₂^-·and·OH.The level of free radical content would affect other indicators,such as GSH,NADH,etc.There was a significant difference in antioxidant enzyme activity between linea alba and rectus abdominis muscle,which was the key factor leading to the difference in free radical content.The redox state in the body could be regulated and maintained by regulating free radical related indexes to ensure the redox balance inside the body.2.The results of HE staining showed that there were some differences in the arrangement of cells inside the linea alba and the rectus abdominis muscle.Masson staining results showed that the linea alba contained a large amount of collagen fibers,while the rectus abdominis muscle contained little.The results of toluidine blue staining showed that there were a certain number of mast cells in the linea alba,but few in rectus abdominis muscle.The results of immunohistochemistry showed that there were a certain number of fibroblasts in the linea alba,but few in rectus abdominis muscle.There was a positive correlation between the number of mast cells and fibroblasts and the content of free radicals.That is to say,the higher content of free radicals,the more numbers of mast cells and fibroblasts in the linea alba.The level of active oxygen radicals in the site could be roughly judged based on the number of mast cells and fibroblasts.3.After DCFH-DA probe was injected into the linea alba,it exhibited a flow phenomenon,while no flow phenomenon was observed after the injection into the rectus abdominis muscle.When the Mito-SOX^TM Red probe was injected into the linea alba,the flow phenomenon will occur,same flow phenomenon was occurred,no flow phenomenon after the injection into the rectus abdominis muscle.The flow phenomenon is directional,from top to bottom,that is,from the end near the head to the end near the tail.The flow velocity of the two kinds of fluorescent probes increased first and then decreased with time.The above results demonstrated that reactive oxygen species were mainly distributed in the linea alba,and that the connective tissue of linea alba could conduct fluorescence probe,which provided a new idea for us to understand the connective tissue.