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Rosemary (Rosmarinus officinalis) leaves possess a variety of bioactivities. Previous studies have shown that the extract of rosemary leaves from supercritical fluid extraction inhibits the expression of inflammatory mediators with apparent dose-dependent responses. In this study, three different extraction conditions (5000 psi at 40, 60, and 80 °C) of supercritical carbon dioxide (SC-CO(2)) toward the extraction of antioxidants from rosemary were investigated. Furthermore, simultaneous comparison of the anti-inflammatory properties between rosemary extract prepared from SC-CO(2) under optimal conditions (5,000 psi and 80 °C) and its purified carnosic acid (CA) using lipopolysaccharide (LPS)-treated murine RAW 264.7 macrophage cells was also presented. Results showed that the yield of 3.92% and total phenolics of 213.5 mg/g extract obtained from the most effective extraction conditions showed a high inhibitory effect on lipid peroxidation (IC(50) 33.4 μg/mL). Both the SC-CO(2) extract and CA markedly suppressed the LPS-induced production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), phosphorylated inhibitor-kappaB (P-IκB), and nuclear factor-kappaB (NF-κB)/p65 in a dose-dependent manner. The five major compounds of verbenone, cirsimaritin, salvigenin, carnosol, and CA existing in the SC-CO(2) extract were isolated by semipreparative HPLC and identified by HPLC-MS/MS analysis. CA was the most abundant recorded compound and the most important photochemical with an anti-inflammatory effect with an IC(50) of 22.5 μM or 7.47 μg/mL presented to the best inhibitory activity on NO production better than that of the 14.50 μg/mL dosage prepared from the SC-CO(2) extract. Nevertheless, the effective inhibition of LPS-induced NF-κB signaling in RAW 264.7 cells from the SC-CO(2) extract extends the potential application of nutraceutical formulation for the prevention of inflammatory diseases.

Diabetic nephropathy progressed to end-stage renal disease (ESRD) is found in type 1 or type 2 diabetes. Oxidative stress is one of the precipitation factors in diabetic nephropathy. Previously, Hibiscus sabdariffa Linnaeus and its polyphenol extracts were found to possess antioxidative effects. This study is aimed to investigate the effect of Hibiscus sabdariffa L. polyphenol extract (HPE) in streptozotocin (STZ) induced diabetic nephropathy. The results show that HPE reduced kidney mass induced by STZ significantly, as well as improving hydropic change of renal proximal convoluted tubules in the rats. HPE also significantly reduced serum triglyceride, total cholesterol and LDL in STZ induced rats. Treatment with HPE significantly increased the activity of catalase and glutathione and reduced lipid peroxidation (thiobarbituric acid-reactive substances, TBARS). The findings of this research show the beneficial effects of HPE on STZ induced diabetic nephropathy including pathology, serum lipid profile and oxidative marker in kidney.