Effect of hyperbaric oxygen treatment on liver hepatocyte damage in oral candidiasis immunosuppressed rats

Background: Liver is an organ that performs a role in metabolism and detoxification of chemical substances. The use of immunosuppressive drugs excessively not only become predisposing factor for oral candidiasis, but also can cause damage and affect liver’s function. Hyperbaric oxygen treatment was known to preserve hepatocytes and prevent liver damage. This study was aimed to investigate the effect of hyperbaric oxygen treatment on liver hepatocyte damage in oral candidiasis immunosuppressed rats. Method: We divided fifteen Wistar rats randomly into three groups: G1 (healthy rats), G2 (immunosuppressed rats with oral candidiasis), and G3 (immunosuppressed rats with oral candidiasis given hyperbaric oxygen). The immunosuppressed condition was made by giving dexamethasone and tetracycline orally for 14 days. Induction of Candida albicans 0.1 cc was conducted on the fourth day. Hyperbaric oxygen treatment was given for five days continuously. The histopathological changes in the liver were measured by counting the amount of normal hepatocytes, pycnotic, karyolysis, karyorrhexis, and score of necrosis area using light microscope with 400x magnification. Result: The amount of pycnotic, karyolysis, and karyorrhexis hepatocytes increased in G2 and decreased significantly in G3, likewise the score of necrosis area. The statistical test using oneway Anova and LSD test showed significant differences (p<0,05) in pycnotic, karyolysis, and karyorrhexis hepatocytes between G1, G2, and G3. Conclusion: The hyperbaric oxygen treatment effect on liver hepatocyte damage in oral candidiasis immunosuppressed rats.


INTRODUCTION
Liver is an organ that performs many different and interrelated functions. The liver plays a role in the metabolism of carbohydrates, proteins, fats, and foreign chemicals. It also become an important key in getting rid of or excreting drugs and other chemical substances into the bile. 1 These chemicals will be detoxified and inactivated so that they become harmless to the body. 2 Hepatocytes play an important role in liver function. These cells make up most of the volume of liver. 3 Drugs and foreign chemicals can cause liver damage by decreasing hepatocytes' resistance and regeneration ability, which in turn can cause harmful effects in liver. 2 Immunosuppression is a condition in which a person is unable to oppose infection because of the decreased immune system. One of the causes of immunosuppression is the use of glucocorticoid drugs which not only have inflammatory effects but also immunosuppression effects. This condition also becomes one of the causes of increasing mycosis infection in the oral cavity, which is called oral candidiasis. Oral candidiasis occurs when some predisposing conditions favor Candida albicans (C. albicans) to become pathogenic. 4,5 The host immune system plays a role in maintaining C. albicans as normal microorganisms commonly found in the oral cavity in a stable condition. 6,7 Decrease in the immune system due to immunosuppression will cause a failure in the elimination of Candida adhesion in oral mucosa, which in turn predisposes to oral candidiasis. 7 However, the use of immunosuppressant drugs continuously and excessively not only become predisposing factor for the occurrence of oral candidiasis, but also affect the liver performance. 8,9 The liver will work harder so that it can damage hepatocyte cells which will then decrease the function of the liver. 2,9 The measurement of the magnitude protective effect of the liver can be done with estimate the enzyme activities, then verifying the liver cell histologically. Liver damage is associated with the necrosis of the liver cells, a decline in the level of tissue glutathione, and the elevation of tissue lipid peroxidation and serum levels of several biochemical parameters. 10 Some previous studies showed that the elevation of transaminase enzyme levels can be used to detect and suggests any inflammation or injury in the liver. Measurement of these enzymes can be used as a tool in identifying the safety of substances that enter and are metabolized by the liver. [11][12][13] Hyperbaric oxygen therapy is known to affect the function of the injured liver. 14 This therapy can increase hepatocyte proliferation in liver damage, and defend hepatocyte necrosis in liver transplantation mice model. The prior study showed that hyperbaric oxygen treatment able to protect hepatocytes from necrosis and apoptosis, as well as to increase sinusoid diameter and microvascular density index in rats model. 15 This treatment has been known to have a protective effect on necrosis of hepatocellular caused by exaggregation of acetaminophen in experimental animals. 16 Hence this study was aimed to investigate the effect of hyperbric oxygen treatment on liver hepatocyte damage in oral candidiasis immunosuppressed rats. are then absorbed and together the blood flow enters the liver to be detoxified. 8 The blood containing these drugs after entering the liver will be in direct contact with the liver sinusoid for detoxification process. The process of detoxification beyond the liver's ability will force the liver to work harder and ultimately result in damage to the liver cells. 2,9 Hepatocyte necrosis emerged due to the excess of toxic compounds that enter the liver or also due to the occurrence of infection in the liver. which then provides benefits for liver cell regeneration. 15 This therapy was also can inhibit the enhancement of lipid peroxidation, and prevent necrosis of the hepatocytes. 15,26 It has known that hyperbaric oxygen treatment can regulate the immune system, which is the activity of phagocytosis and angiogenesis can be stimulated.

Materials
The previous study showed that this therapy can alleviate pro-inflammatory cytokines production in several animal studies, as measured by the subside of inflammatory cell infiltration and tissue edema.
On the other hand, hyperbaric oxygen treatment can enhance the activity of antioxidants, which then provide to diminish inflammation in hepatic injury. 16 The use of immunosuppressant drugs in this study could trigger damage to hepatocytes, which can be seen from the histopathological changes in the liver that showed the degree was mild to moderate. 21 17 This therapy also showed has the effect of repairing liver injury. 15 In the

CONCLUSION
The administration of hyperbaric oxygen treatment 2,4 ATA for five days continuously affect liver hepatocyte damage in oral candidiasis immunosuppressed rats.

ACKNOWLEDGEMENT
The authors wish to acknowledge Universitas Hang Tuah Surabaya for financially supported this research.