Ursolic Acid Inhibits Proliferation and Induces Apoptosis of Cancer Cells In Vitro and In Vivo
Wang Xm, Zhang F, Yang L, et al. Journal of Biomedicine and Biotechnology. Volume 2011 (2011), doi:10.1155/2011/419343
The aims of the study are to explore the effect of ursolic acid (UA) on the growth of gastric cancer cell line BGC-803 and hepatocellular cancer cell H22 xenograft and to understand the mechanism. UA inhibits growth of BGC-803 cells in vitro in dose-dependent and time-dependent manner. Treated with UA in vivo, tumor cells can be arrested to G0/G1 stage. The apoptotic rate was significantly increased in tumor cells treated with UA both in vitro and in vivo. DNA fragmentation was found in BGC-803 cells exposed to UA. UA activated caspase-3, -8, and -9 and down regulated expression of Bcl-2 in BGC-803 cells. The expression of caspase-3 and -8 was elevated in tumor cells from xenograft treated with UA. 18F-FLT PET-CT imaging confirmed tumor model and UA effectiveness. Our results indicated that UA inhibits growth of tumor cells both in vitro and in vivo by decreasing proliferation of cells and inducing apoptosis.
Triterpenes augment the inhibitory effects of anticancer drugs on growth of human esophageal carcinoma cells in vitro and suppress experimental metastasis in vivo.
Yamai H, et al. Int J Cancer. 2009 Aug 15;125(4):952-60. doi: 10.1002/ijc.24433.
The antineoplastic effects of combinations of anticancer drugs (5-fluorouracil, irinotecan and cisplatin) and triterpenes (ursolic acid, betulinic acid, oleanolic acid and a Japanese apricot extract (JAE) containing triterpenes) on esophageal squamous carcinoma cells were examined by the WST-8 (2-(2-methoxy- 4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) assay in vitro and by an animal model in vivo. Triterpenes and JAE showed additive and synergistic cytotoxic effects, respectively, on esophageal squamous carcinoma cells (YES-2 cells) by combinational use of 5-fluorouracil. JAE and 5-fluorouracil induced cell cycle arrest at G2/M phase and at S phase, respectively, and caused apoptosis in YES-2 cells. A new animal model of esophageal cancer causing tumor colonization of the peritoneal cavity and producing bloody ascites was made by injecting YES-2 cells into the peritoneal cavity of a severe combined immunodeficiency mouse. In this model, 5-fluorouracil inhibited colonization of tumor cells in the peritoneum. The addition of JAE to 5-fluorouracil augmented the suppression of experimental metastasis of the peritoneum. The numbers of peritoneal nodules of more than 2 mm in diameter in mice treated with 5-fluorouracil and JAE were less than those in mice treated with 5-fluorouracil alone or JAE alone. These results suggest that triterpenes, especially JAE, are effective supplements for enhancing the chemotherapeutic effect of 5-fluorouracil on esophageal cancer.
Natural sources as potential anti-cancer agents: A review
International Journal of Phytomedicine 3 (2011) 09-26 <http://www.arjournals.org/index.php/ijpm/index>
Ursolic acid increased hepatic uptake of irinotecan as sited in:
Isolation of modulators of the liver specific Organic Anion Transporting Polypeptides (OATPs) 1B1 and 1B3 from Rollinia emarginata Schlecht (Annonaceae)
JPET Fast Forward. Published on August 16, 2011 as DOI:10.1124/jpet.111.184564
Nozawa T, Minami H, Sugiura S, Tsuji A and Tamai I (2005) Role of organic anion transporter OATP1B1 (OATP-C) in hepatic uptake of irinotecan and its active metabolite, 7-ethyl-10-hydroxycamptothecin: in vitro evidence and effect of single nucleotide polymorphisms. Drug Metab Dispos 33:434-439.
The levels of serum vascular endothelial growth factor (VEGF), NO, and proinflammatory cytokines were significantly reduced in ursolic acid–treated animals as was MMP-2 & MMP-9 as sited in:
Antiangiogenic Activity of Ursolic Acid. Integr Cancer Ther June 2010 9: 224-235, first published on May 11, 2010 doi: 10.1177/1534735410367647
Ursolic Acid Inhibits T-Cell Activation through Modulating Nuclear Factor-κB Signaling
Objective: To investigate the effects of ursolic acid (UA) on T-cell proliferation and activation, as well as to examine its effect on nuclear factor-κB (NF-κB) signaling pathway in T cells.
Methods: T-cells isolated from BALB/c mice were incubated with UA at concentrations ranging from 5–30 μmol/L in the presence of phorbol 12-myristate 13 acetate (PMA) or PMA plus ionomycin. The proliferation of T cells was measured by the MTT assay. The expressions of CD69, CD25, and CD71 on T-cell surface were analyzed using flow cytometry. The level of interleukin-2 (IL-2) in the culture supernatant of activated T cells was quantified by enzyme-linked immunosorbent assay (ELISA). The level of phosphorylated IκB-α (p-IκB-α) in total protein and p65, a subunit of NF-κB, nuclear translocation were measured by Western blot analysis.
Results: UA in a dose-dependent manner significantly decreased the proliferation and inhibited the surface expressions of CD69, CD25, and CD71 in murine T lymphocytes upon in vitro activation (P<0.01). Significant reduction of IL-2 production was found in activated T cells treated with UA (P<0.01). The PMA-induced increase in p-IκB-α protein was inhibited, and nuclear translocation of p65 from the cytoplasm was blocked by UA.
Conclusion: UA is a potent inhibitor for T cell activation and proliferation; these effects are associated with the inhibition of NF-κB signalling pathway.
KEYWORDS ursolic acid, T-cell activation, CD69, CD25, CD71, nuclear factor-κB.
Zeng G, Chen J, Liang Q-h, et al. Chin J Integr Med 2012 Jan;18(1):34-39
Basic Introduction and Botanical Info:
Ursolic acid, also known as urson, prunol, micromerol, and malol, is a pentacyclic triterpenoid compound which naturally occurs in a large number of vegetarian foods, medicinal herbs, and other plants. For a long time, it was considered to be pharmacologically inactive. Thus, ursolic acid and its alkali salts (e.g. potassium or sodium ursolates) were exclusively used as emulsifying agents in pharmaceutical, cosmetic, and food preparations. However, upon closer examination, ursolic acid was found to be medicinally active both topically and internally1. Its anti-inflammatory, anti-tumor (skin cancer), and antimicrobial properties make it useful in cosmetic applications.
Definition: ursolic acid:(biochemistry) C30H48O3 A pentacyclic terpene that crystallizes from absolute alcohol solution, found in leaves and berries of plants; used in pharmaceutical and food industries as an emulsifying agent.
Use: Play down blood sugar,oxidation resisting,leave something decrepitude of Human body, antibiotic and anti-inflammation, nourish the liver and promote the function of gallbladder.
Ursolic acid.C30H48O3.CAS No.77-52-1.malol;prunol;Ursolic acid;urson,micromerol,Loquat Leaf extract.Glossy privet leaf Extract.Ursolic acid.25%50%95%HPLC.Pentacyclic Triterpenes photo picture
Ursolic Acid is a pentacyclic triterpenoid. It can be found in many plants including, apple, bilberry, cranberry, elder flower, peppermint, lavender, oregano, thyme, and prunes. Ursolic acid can be used both topically and internally. It is used in cosmetics for its anti-inflammatory and antimicrobial properties. It is also used to treat burns. Ursolic acid has antifungal and antibacterial activities as well.
Distribution: Ursolic acid is present in many plants, including apples, bilberries, cranberries, elder flower, peppermint, lavender, oregano, thyme, hawthorn, prunes.
Like most triterpenoids, ursolic acid is ubiquitous in the plant kingdom. Ursolic acid and its derivatives are constituents of numerous plants which are having diversified phylogenetic origin and taxonomic position. It has been isolated from the protective wax-like coatings of apples, pears, cranberries, prunes, and other fruits. Seaweed are rich in ursolic acid derivatives. Some of the more commonly known medicinal plants containing ursolic acid are shown in Table.
Medicinal plants containing ursolic acid have been used in folk medicine before it was known which constituents were responsible for their therapeutic effectiveness. Contemporary scientific research which led to the isolation and identification of ursolic acid revealed and confirmed that several pharmacological effects, such as, anti-tumor, hepatoprotective, anti-inflammatory (oral and topical), anti-ulcer, antimicrobial, anti-hyperlipidemic and antiviral, can be attributed to ursolic acid.However, its anti-inflammatory (topical), anti-tumor (skin cancer), and antimicrobial properties are pertinent to the cosmetic industry.
Ursolic acid has medicinally action, both topically and internally. Ursolic acid is used in many cosmetic preparations for its anti-inflammatory, antitumor and antimicrobial properties.
Ursolic acid has antibacterial and antifungal activity. Tests have shown that Ursolic acid inhibits the growth of Candida albicans and Microsporium lenosum.
Ursolic acid has anti-inflammatory properties and is used in ointments to treat burns.
Topical application of ursolic acid inhibited TPA-induced initiation and promotion of tumor growth.
Topical application of ursolic acid derived from rosemary extract inhibited TPA-induced tumor initiation and promotion, inflammation, and ornithine decarboxylase activity in mouse skin. Ursolic acid exhibited strong anti-inflammatory activity. It is even more active than of 1 or 2 mmol of ursolic acid along with 5 nmol of TPA for TPA for 20 weeks inhibited the formation of skin tumors per mouse by 45 or 61% respectively.
Lower doses (0.1 or 0.3 mmol) of ursolic acid had a similar inhibitory effect as the higher doses. Twice weekly topical application of 0.1, 0.3, 1, or 2 mmol of ursolic acid along with 5 nmol TPA for 8,12, and 18 weeks reduced the number of skin tumors per mouse by 52-86%, 49-63%, and 44-61%, respectively.
Ursolic acid and its isomer, oleanolic acid have been recommended for skin cancer therapy in China. Topical cosmetic preparations containing ursolic acid/ oleanolic acid have been patented in China for the prevention of topical skin cancer. An ursolic acid/ oleanolic acid ointment inhibited 7,12-dimethylbenz [alpha] anthracene (DMBA)-induced skin cancer in mice. Reportedly, 0% and 3% of mice developed cancer in 15 weeks and 30 weeks, respectively compared to 50% and 90% for the control mice.
Iridoids are common components of plants, but the iridoid glycosides are important active components of only certain Chinese herbs, particularly rehmannia, with which cornus is frequently combined. It is thought that the glucose molecule is cleaved in the digestive system to yield the aglycone as the active compound that has anti-inflammatory actions.
Oleanolic acid (OA) and ursolic acid (UA) are isomers; they have the same chemical formula. These two pentacyclic triterpenes are of interest to scientists because of their numerous biological activities. OA can lower blood sugar, a property attributed to cornus, which is listed among the Chinese herbs used for treating diabetes. UA and OA have liver-protecting and anti-inflammatory effects; cornus is used in some herb formulas for viral hepatitis and the anti-inflammatory effects may be one reason that cornus can be recommended for low back and knee pain associated with aging.
2.Hair Growth Stimulant:
Ursolic acid and its isomer, oleanolic acid, have been used in to nics to enhance hair growth and prevent scalp irritation. Both triterpenoid compounds encourage hair growth by stimulating the peripheral blood flow in the scalp and activating the hair mother cells. They also furnish alopecia-preventing and dandruff-preventing effects.
Beta ursolic acid (triterpenoid sapogenin from the ursan group) inhibited the growth of several strains of staphylococci. Numerous ursolic acid containing plants from the Lamiaceae family exhibited antibacterial/ fungal activity. The minimal inhibitory concentration of Rosmarinus officinalis, Origanum majorana, and Lavandula officinalis were 500, 250, and 500 mg/cm3, respectively. Ursolic acid also inhibited the growth of Microsporium lenosum and Candida albicans at 250 mg/mL.
Ursolic acid treatment improves the health of skin and hair. Ursolic acid and its derivatives form oil-resistant barriers on the skin and hair as they do in the waxy coating of fruits. Ursolic acid has been used to treat photoaged skin because it prevents and improves the appearance of wrinkles and age spots by restoring the skin's collagen bundle structures and its elasticity. Concentrations of ursolic acid ranging from 0.01 to 50 mg have been reported for inclusion in skin treatment preparations
Ursolic acid.C30H48O3.CAS No.77-52-1.malol;prunol;Ursolic acid;urson,micromerol,Loquat Leaf extract.Glossy privet leaf Extract.Ursolic acid.25%50%95%HPLC.Pentacyclic Triterpenes photo picture image
Ursolic acid is a potent anti-inflammatory agent. Thus, it has been recommended for use in burn ointments. It not only inhibits human leucocyte elastase(HLE), but also 5-lipoxygenase and cyclooxygenase activity. Ursolic acid (1.0 mg/ear) inhibited TPA-induced mouse ear edema by 72.4%23. Hirota and coworkers24 determined that 200 mg and 50 mg applications of ursolic acid inhibited 12-O-hexadecanoyl-16-hydroxyphorbol-13-acetate (HPPA) - induced inflammation by 49% and 33%, respectively. In addition, it inhibited concanavalin A (Con A) induced histamine release, which can cause severe inflammation, by 95% at a concentration of 0.001 M.
6.Remembrance and acetylcholinesterase inhibitor:
Soundbite Molecules: Rosemary and Ursolic acid for Remembrance
Simon Cotton, teacher at Uppingham School, takes a look at those compounds that find themselves in the news or relate to our everyday lives.
That's from a Shakespeare play, isn't it?
Yes, in act IV scene v of Hamlet, Ophelia says to Hamlet,,'There's rosemary, that's for remembrance, pray you, love, remember'.
And is there any connection?
One link is that the plant symbolises remembering the dead. In act IV scene v of Romeo and Juliet, rosemary was placed on Juliet's body for her burial ('Stick your rosemary on this fair cor[p]se'). Australians wear a sprig of rosemary on Anzac day to honour the dead soldiers of World War I. The herb grows wild on the Gallipoli peninsula, Turkey, where many Anzacs died during the ill-fated Allied campaign of 1915.
Is there any evidence that rosemary helps memory?
Recent research by Dr Mark Moss and his colleagues at Northumbria University in Newcastle suggests that exposure to rosemary oil can enhance long-term memory by ca 15 per cent.
Which chemicals might be responsible for this effect?
Rosemary contains hundreds of different chemicals. The herb has been used in medicine as a stimulant, possibly owing to its camphor (1) content, while cineole (eucalyptol, 2) and camphor are among the molecules that give the plant microbiocidal properties. Rosemary has long been used as a food preservative. The plant contains antioxidants such as rosemarinic acid (3) and carnosic acid (4). But possibly the most interesting chemical component that affects memory is ursolic acid (5).
Ursolic acid inhibits acetylcholinesterase. This the enzyme catalyses the breakdown of the neurotransmitter chemical acetylcholine, which is important for effective cognition and reasoning. The degeneration of acetylcholine-producing neurons in the brains of people with Alzheimer's disease (AD) has been blamed for the symptom of memory loss associated with AD. Thus, sustaining acetylcholine levels may help delay this memory loss. Ursolic acid has also been shown to slow skin tumour development.
Pentacyclic Triterpenes (PCTs) as Inhibitors of Inflammation:
Pentacyclic triterpenes (PCTs) are a subdivision of an important group of botanical active constituents known as triterpenoids; when complexed with sugar molecules PCTs are called triterpene glycosides, or saponins. Several of the PCTs have inhibitory action for the COX and/or LOX enzymes. Two of the most widely occurring and intensively investigated pentacyclic triterpenes are ursolic acid and its isomer oleanolic acid; their chemical formulas are identical, and their structures differ only by the location of a methyl group (CH3), as displayed at the top of the structure diagrams below. These substances are of considerable interest for potential health benefits, including protective effects against development of cancer, inhibition of existing tumor cells, protection against chemotherapy and radiation therapy adverse effects, anti-inflammatory activity, antioxidant activity, cardiovascular protective effects, and antiviral effects. Several medicinal herbs that contain oleanolic acid, such as ligustrum, forsythia, and swertia, are remedies employed in China for the treatment of hepatitis. Isolated oleanolic acid has already been used in China as a treatment for hepatitis for many years. Ursolic acid is known for its ability to inhibit skin inflammation; in China it is also used as a topical protectant against skin cancer.
These compounds are synthesized in plants from squalene, a compound in which there are no closed carbon rings, but which has the basis for developing five ring (pentacyclic) compounds. Ursolic acid was named after the herb Uva Ursi (bearberry; urs is the root term for bear). Oleanolic acid was named for the olive (olea is the root term for olive), which is rich in this compound; there is a plant family called the Oleanaceae, whose members usually contain oleanolic acid. In general, plants that contain ursolic acid also contain oleanolic acid and vice versa.
Ursolic acid and oleanolic acid form a waxy coating that is found on some plant leaves and fruits. For example, Ligustrum trees (in the Oleanaceae family) are commonly called wax trees because of these components. Ursolic acid is one of the main waxy components on apples that contributes to the smooth, shiny appearance (not to be confused with waxes sometimes added to aid storage of fruits). Squalene (left) is found in some foods, mainly concentrated in oils, such as olive oil, wheat germ oil, and bran oil (about 0.5% of the oil). The most abundant source of squalene is the liver of deep sea sharks (up to 85% squalene in liver oil). Squalene from shark livers is sold in capsules as a dietary supplement that is claimed by proponents to enhance immune functions and reduce inflammation.
The pentacyclic triterpenes are subdivided into several groups, such as the following:
- Oleanane: oleanolic acid, ginsenosides (glycoside form), fusidic acid;
- Ursane: amyrin, ursolic acid, asiatic acid, boswellic acid; and
- Lupane: lupeol, betulin, betulinic acid
Of these, PCTs other than oleanolic and ursolic acid that have received considerable attention are boswellic acid (from frankincense, the resin of Boswellia serrata); asiatic acid (from gotu kola, the aerial part of Centella asiatica), and betulinic acid (from birch trees, the bark of Betula spp.). Their pharmacology is similar to that of oleanolic and ursolic acid. For example, all are being investigated for anti-inflammatory and potential anticancer effects.
Sources of Oleanolic Acid and Ursolic Acid:
Chinese natural products manufacturers are now making high concentrations of oleanolic acid and ursolic acid for formulation in various internal and topical remedies. The technical extracts contain from 25% to 99% pure compound. By contrast, standard herb extracts (e.g., hot water extracts) have up to 2.0-2.5% of ursolic or oleanolic acid (commonly obtained from holy basil, Ocimum sanctum, a common ingredient in Ayurvedic formulas). Plants rich in ursolic acid include Greek sage (Salvia triloba), oleander, rosemary, lavender, and thyme (each having at least 1% ursolic acid in the leaf or whole plant)
Safety and Toxicity for Ursolic Acid:
Notwithstanding its numerous cosmetic properties, ursolic acid is an ideal cosmetic ingredient because its overall toxicity (both chronic and acute) is low. It is not a primary irritant or sensitizer. In fact, it has been termed dermatologically innocuous.
Acute toxicity (LD50):Ursolic acid
LD50-Lethal dose,50 percent death.Intraperitoneal.Rodent-mouse >637mg/kg.
LD50-Lethal dose,50 percent death.Oral.Rodent-mouse.8.33grams(8330 mg)/kg.
Chronic toxicity:Ursolic acid
Hypodermic Injection:Rodent-mouse.3500mg/kg.No Death in 72 hours.
Hypodermic Injection:Rodent-Rat.Dose/Period:0.5 gram/kg.30days.No distinctive effect on body weight, blood, cardiogram, liver,and kidney functions. No pathology changes for important organs,result prove ursolic acid low toxicity and safe enough.
Administration and Dosage:
Clinical use of ursolic acid as an isolate is new, but oleanolic acid, which is nearly identical in structure but apparently milder in action, has been used for some time. Effective daily doses of oleanolic acid, based on its application in treatment of hepatitis, are reported in the range of 75-300 mg/day.
For a standard dried hot water extract of an herb, which may contain about 2% of the PCTs, that dosage corresponds to use of about 4-15 grams of the extract, a large amount. Additional information on dosage of compounds of this type (see: Platycodon and other herbs with triterpene glycosides) indicates that a daily dose of about 300 mg is safe and potentially useful for long term applications and 600 mg is safe at least for short term dosage.
Dr. Acharan Narula describes use of 300-500 mg or oleanolic acid as an intestinal anti-inflammatory and recommends Olenoate, a product of oleanolic acid in capsules of 100 mg each. For ursolic acid, in vitro studies have shown that it is about twice as effective as an anti-inflammatory compared to indomethacin, a potent NSAID that is administered in doses of just 50-100 mg/day; however, when converted to actual human use, the dose of ursolic acid needed may prove higher.
Boswellic acid, another PCT that inhibits 5-LOX, and is used as an anti-inflammatory for arthritis, Crohn's disease, and asthma, is administered in doses of 400 mg 3 times a day of an extract standardized to contain 37.5% boswellic acids, which corresponds to 450 mg/day of boswellic acid.
In general, the PCT dosage for anti-inflammatory effects appears similar to the dosage of NSAIDS.
Oleanolic acid and ursolic acid have been shown to act at various stages of tumor development to inhibit tumor initiation and promotion, as well as to induce tumor cell differentiation and apoptosis. In the two-stage mouse skin carcinogenesis model, the protection of oleanolic acid against 12-Oteradecanoyl phorbol-13-acetate promoted carcinogenesis is associated with inhibition of aberrant gene expression (Oguro et al., 1998). Oleanolic acid derivatives are also effective for acute myeloid leukemia by inducing apoptosis of tumor cells (Konopleva et al., 2004). These triterpenoids and their derivatives are also effective in inhibiting angiogenesis, invasion of tumor cells and metastasis, and emergence as a new class of chemotherapeutics (Ovesna et al., 2004).
*Konopleva, M., Tsao, T., Estrov, Z., Lee, R.M., Wang, R.Y., Jackson, C.E., McQueen, T., Monaco, G., Munsell, M., Belmont, J., Kantarjian, H., Sporn, M.B., Andreeff, M., 2004. The synthetic triterpenoid 2-cyano-3,12-dioxooleana-1 9-dien-28-oic acid induces caspase-dependent and -independent apoptosis in acute myelogenous leukemia. Cancer Research 64, 7927–7935.
*Oguro, T., Liu, J., Klaassen, C.D., Yoshida, T., 1998. Inhibitory effect of oleanolic acid on 12-O tetradecanoylphorbol-13-acetate-induced gene expression in mouse skin. Toxicological Sciences 45, 88–93.
*Ovesna, Z., Vachalkova, A., Horvathova, K., Tothova, D., 2004. Pentacyclic triterpenoic acids: new chemoprotective compounds. Minireview Neoplasma 51, 327–333.