Investigation of the effects of theranekron and vitamin E-selenium on glycerol-induced muscle and kidney damage in rat using pathological and molecular methods

The release of muscle cell contents into the circulation, subsequent to traumatic or non-traumatic damage to the skeletal muscle, results in the development of a uremic syndrome defined as Myoglobinuric Acute Kidney Injury (AKI). Crush syndrome refers to a complex clinical picture encompassing the signs and symptoms that arise from rhabdomyolysis following a crush injury. Large-scale bodily traumas lead to the occurrence of this syndrome. The aim of this study is to investigate the effects of the combined and individual use of the alcoholic extract of Tarantula cubensis (TCE) and the Vitamin E-Selenium combination in Myoglobinuric Acute Kidney Injury A total of 50 female Wistar albino rats were randomly divided into seven groups: 6 rats in each control group and 8 in each experimental group. After 24 hours of water deprivation, the sham control group (S) received 8 mg/kg of physiological saline, divided and injected intramuscularly into both hind limbs. The Glycerol (G), Glycerol + Vitamin E-Selenium (GES), Glycerol + Theranekron (GT), and Glycerol + Vitamin E-Selenium + Theranekron (GEST) groups received 8 ml/kg of 50% hypertonic glycerol was divided and administered intramuscularly (IM) into both hind limbs equally. Following the injection, water was provided ad libitum. At 24 hours after the start of the experiment, treatments were administered as follows and repeated three times (at 24, 72, and 120 hours): 0.3 ml/rat Theranekron subcutaneously, and Vitamin E-Selenium at 100 mg / 1.5 mg/kg intraperitoneally. During the treatment period, the animals were fed ad libitum, and euthanasia was performed on the sixth day. The right kidneys were collected for histopathological examination, and the left kidneys were used for PCR analyses. Histopathological evaluations included scoring the severity of cast and necrosis, desquamation, tubular regeneration, degeneration and dilatation, congestion, mineralization, and glomerular atrophy. Real-Time PCR was used on the right kidneys to determine the expression levels of KIM-1, NGAL, PPARγ, GAPDH, and Cystatin-C. Real-Time PCR analysis results revealed that the glycerol and control groups had statistically significant differences in all parameters. The highest Cystatin-C, NGAL, and KIM-1 expression rates were found in the glycerol group, while KIM-1 levels decreased significantly in the GEST and TK groups, while NGAL levels decreased significantly in the TK group (p<0.05). Changes in Cystatin-C levels were not significantly different between the treatment groups. The lowest PPAR gamma level in the glycerol group was observed to increase significantly in the GEST group (p<0.05), approaching the control group. Histopathological examinations revealed the highest number of pathological findings in the glycerol group, and the GEST group exhibited milder lesions compared to the other treatment groups (p<0.05). This suggested that the combined use of Vitamin E, Selenium, and Theranecron would be more effective in the treatment of rhabdomyolysis. PCR analyses revealed that Theranekron demonstrated a more pronounced effect compared to Vitamin E-Selenium; however, the highest efficacy was observed with the combined application. To fully comprehend the therapeutic potential of these agents, it is recommended that future studies x thoroughly investigate parameters such as optimum dosage, administration route, and treatment duration. Keywords: Rhabdomyolysis, kidney injury, RT-PCR, Theranekron, Vitamin E-Selenium