Protective effects of hydrogen rich saline solution on ventral penile mathieu type flap with penile tourniquet application in rats

Abstract

Penile tourniquet (Pt) application aims to work in a bloodless field in penile surgery. When the tourniquet is released, reperfusion injury occurs with the resumption of blood flow. Molecular hydrogen can easily attach to biomembranes and enter cytosol, mitochondria and other organelles of the cell and convert the formed OH- to H₂O to prevent cell and tissue damage. We investigated the effects of hydrogen rich saline solution (HRSS) on penile Mathieu type flap tissue with Pt application in rats. Thirty-six Wistar-albino male rats were randomly divided into six groups. No operations were performed in the Sham group. Ventral penile Mathieu type flap was prepared and Pt was applied to the root of the penis with a plastic band in other groups. Pt was applied 10 and 30min in the PT1⁰ and PT³⁰ groups. HRSS was injected intraperitoneally (ip) 5ml/kg just before Pt was released in the HRSS1⁰ and HRSS³⁰ groups. In the HRSSB group, HRSS was injected 1h before 10min of Pt application. At the 4th hour of experiments the rats were sacrificed and tissue samples were taken for biochemical and histopathological studies. Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), myeloperoxidase (MPO), malondialdehyde (MDA) and glutathione (GSH) levels were determined in the penile tissue. The results were analyzed with one-way ANOVA and Pearson's Chi-Squared test. Tissue MDA, MPO, IL-6 and TNF-α values were significantly lower in all HRSS groups compared to PT1⁰ and PT³⁰ groups. Tissue GSH levels of HRSS groups were higher compared to PT groups. Histopathologically, inflammation was found to be higher in PT groups compared to HRSS groups. Interestingly, in the HRSSB group with HRSS administration prior to Pt, the damage was less in grade, but not statistically different than the other HRSS groups (p>0.05). In previous studies, damage in histopathological examinations after Pt could only be demonstrated long after tourniquet applications such as 24h and with longer duration of Pt such as 30min. Structural changes in different Pt application times could be demonstrated at 60min by electron microscopy and 48h by light microscopy. In this study, the histopathological effect of Pt application could be demonstrated at the 4th hour after release and HRSS was observed to reduce the damage histopathologically as well as biochemically with its anti-inflammatory and antioxidant effects. It was observed that administration of HRSS either before or following Pt did not cause an alteration statistically. HRSS reduces tissue oxidative stress and inflammation on the flap tissue and has a protective effect in Pt applied to the hypospadias model created with a penile flap.