sennoside Diclofenac ibuprofen and meloxicam are considered
Diclofenac, ibuprofen, and meloxicam are considered among the most commonly used NSAIDs in many countries . Cardiovascular events were reported with these NSAIDs in several clinical studies. In this study, we provide novel information that diclofenac and ibuprofen altered gene expression and may affect the normal ratio of the cardioactive ARA metabolites in the heart and kidneys. Mefenamic sennoside is not widely used, in comparison to other NSAIDs, due to the high rate of side effects such as enteritis and colitis . Several studies showed that it exhibited hepatic and renal toxicities [30,31]. However, there are limited clinical reports regarding mefenamic acid cardiotoxicity . We showed in this study that mefenamic acid produced high alterations in expression of ARA-cyp450 genes in comparison to other NSAIDs used in this study in mouse kidney and heart samples. These molecular findings were in parallel to the abnormal histological alterations induced by mefenamic acid in the tested organs. These data provide an additional step toward understanding the mechanism of mefenamic acid toxicity and its limitations in clinical treatment. Although NSAIDs disturbed the expression of ARA-cyp450 genes toward increasing 20-HETE synthesis in the heart and decreasing it in the kidney, the fold change values in the expression of the targeted genes were different. Furthermore, diclofenac did not have a significant effect, yet other NSAIDs affected the gene expression of hepatic ARA-cyp450 s. This might be due to different chemical properties between the NSAIDs, such as the lipophilicity, which affects the pharmacokinetics  of the drugs in the body and the affinity toward transcriptional receptors and molecular targets . The GATA4 gene is an anti-apoptotic gene which stimulates cardiac cell growth . Down regulation of GATA4 is considered a biomarker of heart failure and was observed in drug-induced heart failure, such as with doxorubicin . In this study, we showed that GATA4 was downregulated more than 2 fold after NSAID treatment and was associated with upregulation of cyp4a12 and cox2 in the heart. These results may indicate that downregulation of GATA4 and upregulation of 20-HETE synthesis genes play a role in the cardiac apoptosis and toxicity induced by NSAIDs. Furthermore, it was shown that 20-HETE induced heart failure through activation of apoptosis in cardiac muscles . Therefore, further studies are needed to investigate the influence of 20-HETE on the gene expression of anti-apoptotic genes, such as GATA4. Induction of cox2 expression is a biomarker of inflammation and apoptosis . Congestive heart failure patients have higher cardiac cox2 expression . In this study, we showed that cox2 gene expression was induced in the heart, but not in the kidney and liver, of NSAID treated mice. This may indicate that NSAID treatment for 14 days induced inflammation in the mouse heart. The compound 20-HETE is considered an inflammatory biomarker and its levels were increased in the inflamed tissues . Our results found that gene expression of 20-HETE synthesizing cyp4a12 was associated with cox2 expression in the heart, which suggests a possible role of cyp4a12 expression in cardiac inflammation induced by NSAID treatment. ANP and BNP are considered as biomarkers for heart failure and myocardial infarction and they were downregulated in the drug-induced toxicity . However, we found in this study that 14 day NSAID administration dwonregulated ANP and BNP but did not reach the statistical significance (p value > 0.05). This might indicate that the effect of NSAIDs on the expression of ANP and BNP genes is time- or dose-dependent. Although 20-HETE is considered cardiotoxic in the cardiac muscle, it increases diuresis in the kidney and decreases blood pressure . Several studies showed a correlation between elevated blood pressure and reduction of 20-HETE synthesis in the kidney [40,41]. In addition, loss of function genetic variants (T8590C) in the human CYP4 A11 gene were associated with hypertension . In the current study, we showed that cyp4a12 was decreased in the kidney after NSAID treatment, which might result in the decline of 20-HETE synthesis in the kidney. On the other hand, the expression of the antidiuretic EETs cyp2c29 gene  was highly induced by more than 6 fold in the kidney after diclofenac and mefenamic acid administration. Collectively, these molecular alterations may indicate that the ratio of synthesis of EETs to 20-HETE was increased in the kidney and hence the diuretic effect was decreased.