Ursolic acid rosemary

Huang MT, Ho CT, Wang ZY, et al. Inhibition of skin tumorigenesis by rosemary and its constituents carnosal and ursolic acid. Cancer Res 1994 54 701-708. 

Supplementation of diets for 2 weeks with rosemary extract (0.5%) but not camosol (1.0%) or ursolic acid (0.5%) resulted in a significant decrease in the in vivo formation of rat mammary DMBA-DNA adducts compared to the control. When injected intraperitoneally for 5 days at 200 mg/kg body wt rosemary and camosol, but not ursolic acid, significantly inhibited mammary adduct formation and were associated with a significant decrease of 74 and 65%, in the number of DMBA-induced mammary adenocarcinomas per rat (Singletary et al 1996). 

Main components 1.5-2.5% essential oil, rosemarinic acid (see rosemary), flavonoids [266, 267], tannins, oleanolic and ursolic acid. 

Labiatae Basil, marjoram, mint, thyme, rosemary, dill, oregano, sage Mono-, diterpenes, flavonoids, rosmarol, ursolic acid, phenolic derivatives... 

When the rosemary extract was administered (i.g.) at lOOmg/kg/day for 5 consecutive days, the number and area of diethylnitrosamine-induced GST placental-form-positive (GST-P) hepatocellular foci were reduced in male F344 rats. A methanol extract of the leaves of rosemary was evaluated for its effects on tumor initiation and promotion in mouse skin carcinogenesis. Topical apphcation of the rosemary extract to mouse skin attenuated the covalent binding of BaP to epidermal DNA and inhibited tumor initiation by BaP and DMBA. Application of rosemary to mouse skin also inhibited TPA-induced ODC activity, inflammation, hyperplasia, and tumor promotion. Likewise, topical application of camosol or ursolic acid isolated from rosemary inhibited TPA-induced ear inflanunation, ODC activity, and tumor promotion. ... 

Rosemary (Rosmarinus officinalis) contains flavonoids, phenols, volatile oil and terpenoids. Topical application of rosemary extract, carnosol or ursolic acid to mouse skin inhibited the covalent binding of benzo[a]pyrene to epidermal DNA (Huang et al. 1994), tumour initiation by 7,12-dimethylbenz [ajanthracene (Singletary and Nelshoppen 1991), 12-0-tetradecano)dphorbol-13-acetate-induc-ed tumour promotion, ornithine decarboxylase (EC 4.1.1.17) activity and inflammation. Carnosol showed potent antioxidative activity in a,a-diphe-nyl-P-picrylhydrazyl free radicals scavenge and DNA protection from Fenton reaction (Lo et al. 2002). 

The antioxidant potential of plant materials, such as rosemary, also comes from some triterpenic acids such as betulinic, oleanolic and ursolic acids, triterpenic alcohols (P-amyrin and others, see Section 3.7.4.1.1) and derived saponins (see Section 10.3.2.2). Usually low antioxidant activity (up to temperatures around 180 °C) is... 

Camosol and ursolic acid were isolated from rosemary as described (5). Camosic acid and rosmarinic acid were isolated from the ground dried leaves of rosemary, sequentially, by hexane and n-butanol extractions. The final products were purified by column chromatography on silica gel (2). LPS (lipopolysaccharide, Escherichia coli 026 B6), sulfanilamide and naphthyl-ethylenediamine dihydrochloride were purchased from Sigma Chemical Co. (St. Louis, MO). Isotopes were obtained from Amersham (Arlington Heights, IL). RT-PCR reagents were purchased from Promega (Madison, WI). Polynucleotide kinase was purchased from Pharmacia (Piscataway, NJ). 

The major rosemary phytochemicals including camosol, carnosic acid, ursolic acid and rosmarinic acid (Figure 1) were separated by HPLC as described in Materials and Methods and illustrated in Figure 2. The retention time of these compounds were carnosic acid, 19.5 min camosol, 7.7 min ursolic acid, 13 min and rosmarinic acid, 2-12 min. It seemed that rosmarinic acid was not resolved well in this system. The relative amounts of these compounds in rosemary extracts were similar to previous findings (5) as 16.5-19.2% ursolic acid 3.8-4.6% camosol 0.1-0.5% carnosic acid and trace amount of rosmarinic acid, respectively. 

Cells were treated with rosemary phytochemicals (0-20 pM) for 24 h. Cell viability was assayed using ATP-Lite -M non-radioactive cell proliferation assay kit and luminescence intensity measured by Topcount Microplate Scintillation and Luminescence Counter. The results indicated that ursolic acid showed strong inhibitory effect on the viability of RAW 264.7 cells camosol and carnosic acid showed moderately activity, while rosmarinic acid was less active than camosol and carnosic acid (Figure 3). 

Figure 1. Structure of rosemary phytochemicals, (A) carnosic acid (B) carnosol (C) rosmarinic acid (D) ursolic acid.

Figure 2. High performance liquid chromatographic separation of rosemary phytochemicals. The chromatographic conditions were as described in the Materials and Methods. The abbreviations are CL, carnosol VA, ursolic acid CA, carnosic acid RA, rosmarinic acid.

Figure 4. Effects of rosemary phytochemicals on LPS-induced nitrite formation in RAW 264.7 cells. Cells were treated with or without indicated concentrations of tested compounds [carnosic acid (CA), carnosol (CL), rosmarinic acid (RA), ursolic acid (UC)] and LPS (1 pg/mL) for 24 h. Amounts of nitrite released to culture medium was determined by Griess reagent and read at OD 550 nm with a sodium nitrite standard curve. Data represent means SE of triplicate tests.

Like many other antioxidants, rosemary and its polyphenols possess not only antioxidative activities, but also antitumorigenic activities. Huang et al. (5) investigated the inhibitory effects of rosemary extract, carnosol and ursolic acid on tumor formation in mouse skin. They found that topical application of rosemary inhibits B(a)P- and DMBA-induced initiation of tumor and TPA-induced tumor promotion in DMBA-initiated mice. Carnosol and ursolic acid were found to be strong inhibitors of TPA-induced inflammation, ornithine decarboxylase activity and tumor promotion in mouse skin (5). It was suggested that carnosol acted like other nonsteroidal phenolic anti-inflammatory agents, such as curcumin, which inhibited the metabolism of arachidonic acid. 

The aim of this study is to elucidate whether camosic acid, camosol, rosmarinic acid and ursolic acid have any effects on apoptotic induction, since ursolic acid was previously reported to possess this effect previously (28). We used HL-60 cells to investigate the molecular mechanisms involved. We examined the effects of these rosemary phytochemicals on DNA fragmentation, activation of caspases, altering the mitochondrial function, ROS generation and releasing of cytochrome c from mitochondria. In the present study, we have demonstrated camosic acid, camosol, and ursolic acid induced apoptosis in HL-60 ceils and activated caspase-3 and caspase-9 via provoking the release of cytochrome c. 

Camosol, camosic acid, rosmarinic acid and ursolic acid were isolated from rosemary (/). Their sfructures are shovm in Figure 1. RNase A, proteinase K and propidium iodide were purchased from Sigma (St. Louis, MO). Substrates for caspase-1, Ac-YVAD-AMC casppase-3, Ac-DEVD-AMC caspase-8, Ac-lETD-AMC and caspase-9, Ac-LEHD-AMC were obtained from AnaSpec Inc. (San Jose, CA). DiOC6 (5) and DCFH-DA were obtained from Molecular Probes (Eugene, OR). 

The inhibition of skin carcinogenesis by rosemary and its constituents, camosol and ursolic acid, has been demonstrated by Huang et al. (/). Furthermore, the suppression of rat mammary tumorigenesis induced by DMBA has been demonstrated by Singletary et al. (29). Meanwhile, the formation of mammary DMBA-DNA adducts in vivo was dose-dependently inhibited by camosol and ursolic acid (29). Rosemary components have the potential to decrease activation and increase detoxification of benzo(a)pyrene, identifying them as promising chemopreventive agents (30). 

Main components 1-3% essential oil, rosemary acid (primarily responsible for the antioxidating effect of sweet marjoram), ursolic and oleanolic acid, phenols like arbutin [152], flavonoids [153, 154]. 

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