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EXPERIMENTAL JUSTIFICATION OF NEW WAY OF PHARMACOLOGICAL CORRECTION FOR CONTACT FROSTBITE USING DSLET OPIOID PEPTIDE AND SEROTONIN ADIPINATE TO ENHANCE SURGYCAL TREATMENT

The severity of complications and a large percentage of disability related to the occurrence cold injury lead to medical, social and economic problems. Considering the contact of cold injury as a variety of acute ischemia with the subsequent formation of early and late postischemic disorders as causes of disorders of blood circulation in the affected segments can be considered a disorder of the oxidant-antioxidant status and cytokine, systemic inflammation, reperfusion syndrome, the development of endothelial dysfunction, which ultimately leads to multiple organ failure. In the course of the study when assessing the microcirculatory changes caused by the simulation of frostbite, the most pronounced effect of the combined treatment of Serotonin adipinate 4.5 mg/kg and opioid peptide DSLET 10 μg/kg compared with monotherapy with the studied drugs was estimated. The most effective correction of indices of oxidant-antioxidant protection observed with the combination of necrectomy with a combination of Serotonin adipinate and opioid peptide DSLET, which was confirmed by approximating the indicators of malondialdehyde, superoxide dismutase, finite stable metabolites of nitrogen oxide, total antioxidant activity and catalase to the values of intact animals on the 14th day of the experiment and reducing the level of proinflammatory cytokines (interleukin-6, tumor necrosis factor-alpha) in 2 times (p < 0.05) compared to the Control group. The combined use of Serotonin adipinate 4.5 mg/kg and opioid peptide DSLET of 10 µg/kg in combination with active surgical tactics (necrectomy) helps reduce the severity and prevalence of alternative changes, accelerate the formation of granulation shaft, a more intense development of reparative processes in the simulation of cold injury compared with the monotherapy. The combined use of investigational drugs in combination with active surgical tactics led to the maximum enhancement of the survival of the animals with contact frostbite in the experiment.

Иллюстрации

Figure 1: Microcirculation variation curve in the necrosis-bordering area in Control group 1 and Study groups 1-3 under conditions of experimental frostbite

NOTE:            x – at p<0.05 as compared with the Intact group; y – at p<0.05 as compared with the Control group 1. 

Figure 2 (a-b). Influence of Serotonin adipinate (4.5 mg/kg), DSLET (10 µg/kg) and the impact of their combined use on biochemical indicators of rats’ blood serum in simulation of contact frostbite

NOTE:            X - at p<0.05 as compared with the Intact group; Y - at p<0.05 as compared with the Control group 1.

 Figure 2 (c-d). Influence of Serotonin adipinate (4.5 mg/kg), DSLET (10 µg/kg) and the impact of their combined use on biochemical indicators of rats’ blood serum in simulation of contact frostbite

NOTE:            X - at p<0.05 as compared with the Intact group; Y - at p<0.05 as compared with the Control group 1.

  Figure 2 (e-f). Influence of Serotonin adipinate (4.5 mg/kg), DSLET (10 µg/kg) and the impact of their combined use on biochemical indicators of rats’ blood serum in simulation of contact frostbite

NOTE:            X - at p<0.05 as compared with the Intact group; Y - at p<0.05 as compared with the Control group 1.

 Figure 2 (g). Influence of Serotonin adipinate (4.5 mg/kg), DSLET (10 µg/kg) and the impact of their combined use on biochemical indicators of rats’ blood serum in simulation of contact frostbite

NOTE:            X - at p<0.05 as compared with the Intact group; Y - at p<0.05 as compared with the Control group 1.

Figure 3. Estimation of level of IL-6 (a) and TNF-α (b) in the blood of experimental animal in Study groups 4-6 versus Control group 2 and intact animals

NOTE:            X - at p<0.05 as compared with the Intact group; Y - at p<0.05 as compared with the Control group 2.

 

Figure 4a. Morphological changes of tissues in the area of damage in application of the combination of Serotonin adipinate and DSLET (Study group 3) in experimental contact freezing.

NOTE: Development of fibrinoid connective tissue necrosis in the area of damage, heterogeneity of alteration, presence of survived foci of derma on the 3-rd day. Van Gieson's method. X 100

 

 

Figure 4b. Morphological changes of tissues in the area of damage in application of the combination of Serotonin adipinate and DSLET (Study group 3) in experimental contact freezing.

NOTE: Significant demarcation zone completely surrounds the area of necrosis on 3–rd day. Haematoxylin-eosin. X 100

Figure 4c. Morphological changes of tissues in the area of damage in application of the combination of Serotonin adipinate and DSLET (Study group 3) in experimental contact freezing.

NOTE: Newly formed connective tissue fibers in the forming scar in the area of damage on the 14-th day. Van Gieson's method. X 400. 

Figure 4d. Morphological changes of tissues in the area of damage in application of the combination of Serotonin adipinate and DSLET (Study group 3) in experimental contact freezing.

NOTE: Proliferation of epithelium on the surface of mature connective tissue in the area of damage on the 14th day. Haematoxylin-eosin. X 200. 

Figure 5a. Morphological changes of tissues in the area of damage after necrectomy in case of the combination of Serotonin adipinate and DSLET (Study group 6) in experimental contact freezing.

NOTE: Accumulation of connective tissue fibers in the young connective tissue in the area of damage, 7-th day. Haematoxylin-eosin. X 400.

 

Figure 5b. Morphological changes of tissues in the area of damage after necrectomy in case of the combination of Serotonin adipinate and DSLET (Study group 6) in experimental contact freezing.

NOTE: Connective tissue in the developing scar in the area of damage, 14-day. Haematoxylin-eosin. X 400.

Figure 6. The survival rate of experimental animals on the background of treatment with Serotonin adipinate 4.5 mg/kg, DSLET 10 μg/kg and their combination without necrectomy (a) and in combination with necrectomy (b) on the 14th day of modeling contact frostbite

NOTE: X – at p<0.05 in comparison with the Control group.

DOI: 10.18413/2313-8971-2016-2-2-3-19
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