Evaluation of Anti-Inflammatory Agent In Cancer-Induces Cachexia

Abstract

BACKGROUND AND OBJECTIVE: Cancer-associated cachexia is a condition characterized by weight loss, anorexia, decrease in muscle mass, lipolysis, fatigue, and elevated energy expenditure. Muscle atrophy in cancer is marked by chronic systemic inflammation, which reduces the effectiveness of treatment and raises the risk of cancer-related mortality. The inflammatory cytokines are released by tumors, which promote protein breakdown and reduce protein production. Levels of inflammatory cytokines like TNF-α, IL-6, IL-1, and INF-ϒ are found elevated in cachectic patients which activates pathways of protein degradation and suppresses protein synthesis resulting in muscle wasting. Till now, few interventions are used in clinical studies to treat the disease which focus on targeting inflammatory cytokines, myostatin, and metabolism modulators, and to reverse anemic, anorexia, nausea, depression, loss of body weight, and fatigue-like conditions. Cyclooxygenase (COX) enzyme is having a role in cancer cachexia. Arachidonic acid is synthesized by COX to induce the production of prostaglandins (PGs). PGs play a crucial function in inflammation. PGs production elevates the levels of IL-6 and MuRF-1, which in turn promotes protein breakdown and inhibits synthesis. A result of this imbalance is muscle atrophy. Hence by targeting COX, NF-κB and inflammatory cytokines can be a potential target to treat cancer cachexia. Non-steroidal anti-inflammatory agents (NSAIDs) are inhibitors of COX enzyme. Nabumetone is reported of having anti-inflammatory action by inhibiting COX enzyme and is used in rheumatoid and osteoarthritis. Studies have also revealed nabumetone’s activity on the NF-κB pathway. Despite this, there is no information available on nabumetone's impact on cancer cachexia. Therefore, the main objective of this study was to investigate the role of Nabumetone, an NSAID, in cancer cachexia and to develop a treatment strategy for cancer cachexia. MATERIALS AND METHOD: Female C57/BL6 mice, of 10-12 weeks of age, weighing 20-30 gm were taken for the study and were maintained under controlled conditions of temperature (25° ± 2° C), humidity (55 ± 5%), and 12 hours light-dark cycle. Standard laboratory rat chow and UV-filtered water were provided ad libitum. B16f10 (~5x10 6 cells) were injected subcutaneously into each C57/BL6 mice. Tumor volume was recorded with a digital caliper every two days, measuring the higher and lower diameters of tumors. Once a tumor volume of 1500 mm3 was achieved mice were treated with treatment drugs for 28 days. The evaluation of parameters was performed which includes tumor markers, body mass parameters, inflammatory markers, markers for the metabolism of carbohydrates and lipids, markers for wasting of skeletal muscle, and cardiac complications. RESULTS: Body mass markers: Animals with cancer cachexia showed a marked decrease in body weight, food intake, carcass weight, and delta lean mass when compared to normal control animals. We noted that there was a noteworthy increase in body weights, food intake, carcass weight, and delta lean mass in nabumetone-treated animals in comparison to cancer-cachexia control animals. Lipid metabolism marker: Cancer cachexia animals exhibit impaired lipid metabolism. Nabumetone-treated animals showed improvement in lipid metabolism. Cholesterol, triglycerides, VLDL, LDL, and HDL were measured as markers of lipid metabolism. Carbohydrate metabolism marker: A significant increase in serum glucose levels was observed in disease-control animals. After the treatment with nabumetone, there was a substantial decrease in serum glucose levels. Inflammatory markers: B16F10 cell line-induced cancer cachexia animals were having elevated levels of TNF-α and IL-6 compared to normal control animals. Treatment with nabumetone significantly decreased levels of inflammatory cytokines. Skeletal muscle wasting parameters: The weights of skeletal muscles such as Tibialis anterior, EDL, gastrocnemius, soleus, and quadriceps muscles were significantly lower in disease control group relative to normal control. Nabumetone treatment produced a significant increase in the weight of tibialis anterior, EDL, and quadriceps muscles when compared to cancer cachexia control animals. A decrease in locomotor activity was noted in disease-control animals. After receiving treatment increase in locomotor activity was reported. Tumor parameters: Treatment with nabumetone does not show a significant decrease in tumor weight and tumor volume compared to the disease-control animals. Cardiac complication markers: A decrease in heart weight and elevated levels of CK-MB was observed in disease control group. After receiving treatment with nabumetone a significant increase in heart weight and decrease in CK-MB levels was found. Histological analysis: Histological analysis of H&E staining showed disarrangement of skeletal muscle and cardiac muscle fibers in cancer-cachexia animals compared to the control groups. Nabumetone-treated animals exhibited correctly organized muscle fibers. Small lipid droplets were seen in the disease control group. Treatment with nabumetone showed larger lipid droplets compared to cachectic animals. Picro-Sirius red and Masson-trichome staining were carried out to determine collagen content in skeletal muscles. Disease control animals show a decrease in collagen content in both staining. Nabumetone treatment showed an increase in collagen content compared to the disease control group. CONCLUSION: Our data suggest that NSAID nabumetone produces a beneficial effect on cancer cachexia by increasing body weight, food intake, carcass weight, and lean body mass. Decrease in serum glucose, inflammatory cytokines, and CK-MB were observed after treatment with nabumetone was given. Wasting of cardiac muscle, skeletal muscles and fat tissue was improved after giving nabumetone. Thus, nabumetone can help cancer patients overcome cachexia.