[CANCER RESEARCH 48, 5941-5946, November 1, 1988] Inhibitory Effect of Curcumin, Chlorogenic Acid, Caffeic Acid, and Ferulic Acid on Tumor Promotion in Mouse Skin by 12-O-Tetradecanoylphorbol-13-acetate
Mou-Tuan Huang, Robert C. Smart, Ching-Quo Wong, and Allan H. Conney Department of Chemical Biology and Pharmacognosy, College of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08855-0789 [M-T. H., A. H. C.]; Roche Research Center, Hoffmann-La Roche Inc., Nutley, New Jersey 07110 [M-T. H., R. C. S., C-Q. W., A. H. C.]; and Toxicology Program, North Carolina State University, Raleigh, North Carolina 27695-7633 [R. C. SJ
ABSTRACT also evaluated the effects of the related compounds chlorogenic acid, caffeic acid, and ferulic acid as potential inhibitors of The effects of topically applied curcumin, chlorogenic acid, caffeic acid, and ferulic acid on 12-O-tetradecanoylphorbol-13-acetate (TPA)- tumor promotion. induced epidermal ornithine decarboxylase activity, epidermal DNA syn thesis, and the promotion of skin tumors were evaluated in female CD-I MATERIALS AND METHODS mice. Topical application of 0.5, 1, 3, or 10 iano\ of curcumin inhibited by 31, 46, 84, or 98%, respectively, the induction of epidermal ornithine Materials. TPA was purchased from CRC Inc., Chanhassen, MN. decarboxylase activity by 5 nmol of TPA. In an additional study, the DL-['"C]Ornithine (58 Ci/mmol) and [3H]thymidine (5 Ci/mmol) were topical application of 10 ¿tinolofcurcumin, chlorogenic acid, caffeic acid, purchased from Amersham Corp., Arlington Heights, IL. fra/w-Reti- or ferulic acid inhibited by 91, 25,42, or 46%, respectively, the induction noic acid was obtained from Hoffmann-La Roche Inc., Basle, Switzer of ornithine decarboxylase activity by 5 nmol of TPA. The topical land. Fluocinolone acetonide, nonradioactive DL-ornithine, curcumin, application of 10 iimol of curcumin together with 2 or 5 nmol of TPA inhibited the TPA-dependent stimulation of the incorporation of |'I l|- chlorogenic acid, ferulic acid, and caffeic acid were purchased from the Sigma Chemical Co., St. Louis, MO, and DMBA was purchased from thymidine into epidermal DNA by 49 or 29%, respectively, whereas lower Calbiochem-Behringer, San Diego, CA. Aquasol was purchased from doses of curcumin had little or no effect. Chlorogenic acid, caffeic acid, New England Nuclear, Boston, MA. Acetone and dimethyl sulfoxide and ferulic acid were less effective than curcumin as inhibitors of the TPA-dependent stimulation of DNA synthesis. Topical application of 1, were purchased from Burdick & Jackson Laboratories, Muskegon, MI. Arachidonic acid was purchased from Nu-Chek Prep Inc., Elysian, MN. 3, or 10 fimol of curcumin together with 5 nmol of TPA twice weekly for Animals. Seven-week-old female CD-I mice were purchased from 20 weeks to mice previously initiated with 7,12-dimethylbenz[a]anthra- cene inhibited the number of TPA-induced tumors per mouse by 39, 77, Charles River Laboratories, Kingston, NY, and kept in our animal facility at least 1 week before use. Mice were fed a Purina Laboratory or 98%, respectively. Similar treatment of mice with 10 nmol of chloro Chow 5001 diet ad libitum (Ralston-Purina Co., St. Louis, MO) and genic acid, caffeic acid, or ferulic acid together with 5 nmol of TPA kept on a 12-h light, 12-h dark cycle. Mice were provided drinking inhibited the number of TPA-induced tumors per mouse by 60, 28, or water ad libitum. The dorsal region of each mouse was shaved with 35%, respectively, and higher doses of the phenolic acids caused a more electric clippers at least 2 days before treatment with TPA or DMBA. pronounced inhibition of tumor promotion. The possibility that curcumin Only mice that did not show signs of hair regrowth were used. All could inhibit the action of arachidonic acid was evaluated by studying the compounds were applied to the dorsal shaved area of 8- to 9-week-old effect of curcumin on arachidonic acid-induced edema of mouse ears. The mice in 200 ß\acetoneor 200 ^1 of acetone:DMSO (90:10). topical application of 3 or 10 ¿tmolof curcumin 30 min before the Ornithine Decarboxylase Assay and Preparation of Epidermal I lo- application of 1 /imol of arachidonic acid inhibited arachidonic acid- mogenates. The preparation of epidermal homogenate and the deter induced edema by 33 or 80%, respectively. mination of ornithine decarboxylase activity were done as described previously (18). Mice were treated topically with 200 /il of acetone, INTRODUCTION with TPA in acetone, or with curcumin and TPA together in acetone. In experiments with chlorogenic acid, caffeic acid, and ferulic acid, The ground dried rhizome of the plant Curcuma longa Linn mice were treated with 200 n\ of acetone:DMSO (90:10), with TPA in has been used for centuries as a naturally occurring medicine acetone:DMSO, or with the test compound and TPA together in for the treatment of inflammatory and other diseases (1,2), and acetone:DMSO. Five h after treatment, the mice were sacrificed by it has also been used as a coloring agent and spice in many cervical dislocation, and the dorsal area of the skin was removed. In order to remove the epidermis from the dermis, the skins were plunged foods. The powdered rhizome is commonly called turmeric. into a 58°Cwater bath for 30 s, and then the skins were immediately Curcumin (diferuloylmethane; Fig. 1) has been identified as the major pigment in turmeric, and this substance has also been submerged in an ice water bath as described by Slaga et al. (19). The epidermis was removed from the dermis by gentle scraping and placed used as a spice, food preservative, and yellow coloring agent in 1 ml of 50 mM potassium phosphate, pH 7.7, buffer containing 2 (3-5). Curcumin, which is widely used in curry, is responsible HIMdithiothreitol and 0.1 mM EDTA. The epidermis was homogenized for the yellow color of curry. Turmeric contains 1-5% curcumin on ice for 30 s with a Tekmar polytron tissumizer at full setting. The while the curcumin content of turmeric oleoresin is approxi epidermal homogenate was centrifuged at 11,000 x g for 30 min at 4°C mately 40% (6). and the supernatant fraction was removed and stored overnight at Recent studies have indicated that several compounds that —20°Cpriorto the determination of ornithine decarboxylase activity possess antioxidant or antiinflammatory activity inhibit TPA1- as described earlier (18). Protein was determined by the method of induced tumors on mouse skin (7-12). Since curcumin has been Lowry et a/., using bovine serum albumin as the standard (20). reported to possess both antioxidant and antiinflammatory Quantitation of Epidermal DNA Synthesis. Mice were treated topi activity (13-17), we have evaluated the effect of curcumin on cally with 200 ii\ acetone, TPA in acetone, or curcumin and TPA TPA-induced tumor promotion on mouse skin, and we have together in acetone. Acetone:DMSO (90:10) was the solvent for exper iments with chlorogenic acid, caffeic acid, and ferulic acid. At 18 h after treatment, 10 ^1 of [3H]thymidine solution (5 Ci/mmol, 1 mCi/ Received 5/6/88; revised 7/22/88; accepted 7/29/88. The costs of publication of this article were defrayed in part by the payment ml) were injected i.p. into each mouse, and 40 min later the mice were of page charges. This article must therefore be hereby marked advertisement in sacrificed by cervical dislocation. The isolation of epidermal DNA and accordance with 18 U.S.C. Section 1734 solely to indicate this fact. determination of the incorporation of [3H]thymidine into DNA was 'The abbreviations used are: TPA, 12-O-tetradecanoylphorbol-13-acetate; DMBA, 7,12-dimethylbenz[a]anthracene; DMSO, dimethyl sulfoxide; HETE, done as previously described (18). hydroxyeicosatetraenoic acid; HPETE, hydroperoxyeicosatetraenoic acid. Tumor Studies on Mouse Skin. The dorsal region of 7-week-old 5941
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CH cate that curcumin is about 10-fold more active than the phe OO II II HO CH= CH-C-CH2-C-CH = CH nolic acids. The results of an additional study indicated that an i.p. injection of curcumin could inhibit the induction of orni CURCUMIN thine decarboxylase activity in mouse epidermis that occurred after the topical application of TPA. The i.p. injection of 40 or 120 /¿molof curcumin (dissolved in dimethyl sulfoxide) into HOOC-CH= CH mice l h before the topical application of 5 nmol TPA inhibited the induction of epidermal ornithine decarboxylase activity by FERULIC ACID 46 or 86%, respectively (data not shown). During the course of this study, it was found that curcumin in the vehicle precipitated out of solution in the abdominal cavity and thus may not have
HOOC-CH=CH been completely bioavailable. Curcumin could be seen as yellow flecks in the abdominal cavity throughout the experimental
CAFFEIC ACID period of 6 h. Effect of Curcumin, Chlorogenic Acid, Caffeic Acid, and Fer ulic Acid on TPA-induced DNA Synthesis in Mouse Epidermis.
H OOC-CH = C Topical application of 1, 3, or 10 nmo\ of curcumin together
HOOCJ 1 H with 2 nmol of TPA inhibited the TPA-induced increase in DNA synthesis by 4, 14, or 49%, respectively (Table 2, experi ment 1). The topical application of fluocinolone acetonide (2 nmol), a known inhibitor of TPA-induced DNA synthesis, CHLOROGENICACID inhibited the TPA-induced increase by 86%. Topical applica Fig. 1. Structures of curcumin, ferulic acid, cafTeicacid, and chlorogenic acid. tion of 1, 3, or 10 ¿¿molofcurcumin simultaneously with 5 female CD-I mice was shaved with an electric clipper. Two days later nmol of TPA inhibited the TPA-induced increase in the incor poration of [3H]thymidine into epidermal DNA by 3,4, or 29%, groups of 30 mice were treated topically with 200 nmol DMBA in 200 til acetone, and control mice received 200 ill of acetone alone. After 1 respectively (Table 2, experiment 2). Topical application of 20 week, mice were treated topically with 200 til of acetone, 5 nmol TPA, /¿molof chlorogenic acid, caffeic acid, or ferulic acid together or 5 nmol TPA applied simultaneously with curcumin in 200 ^1 of with 5 nmol of TPA inhibited the TPA-induced increase in the acetone twice weekly for 20 weeks. In another tumor study designed to incorporation of [3H]thymidine into epidermal DNA by 7, 33, evaluate the effect of chlorogenic acid, cafTeic acid, and ferulic acid on or 15%, respectively. TPA-induced tumor formation, acetone:DMSO (90:10) was used as Effect of Curcumin, Chlorogenic Acid, Caffeic Acid, and Fer the solvent because chlorogenic acid at the doses examined was not ulic Acid on TPA-induced Tumor Promotion in Mouse Epider completely soluble in acetone. Tumors of at least 1 mm in diameter mis. CD-I mice initiated with 200 nmol of DMBA and pro were counted and recorded once every 2 weeks, and the results are expressed as the average number of tumors per mouse and percentage moted with 5 nmol of TPA twice weekly for 20 weeks developed of tumor-bearing mice. Control mice that were first initiated with 200 an average of 16.4 tumors/mouse. Topical application of 1, 3, nmol DMBA and then treated with 3 //mol curcumin in acetone or or 10 ttmol of curcumin with 5 nmol of TPA twice a week for acetone alone twice weekly for 20 weeks also were examined. Likewise, 20 weeks inhibited the number of skin tumors per mouse by control mice treated with acetone alone in place of DMBA and then 39,77, or 98%, respectively, and the percentage of animals with treated twice weekly with 5 nmol TPA in 200 M!acetone for 20 weeks tumors was decreased by 21, 66, or 82%, respectively (Fig. 2). were also included in the study. Additional groups of mice were initiated with DMBA and then treated with acetone or 3 /¿molofcurcumin twice weekly for 20 weeks. None of these animals developed tumors, indicating that RESULTS curcumin was not a tumor promoter. In a second experiment, Effect of Curcumin, Chlorogenic Acid, Caffeic Acid, and Fer mice that were initiated with 200 nmol of DMBA and promoted ulic Acid on TPA-induced Ornithine Decarboxylase Activity in with 5 nmol of TPA twice weekly for 19 weeks developed an Mouse Epidermis. The effect of topically applied curcumin, average of 6.2 tumors/mouse. In this experiment, topical ap chlorogenic acid, caffeic acid, or ferulic acid on TPA-induced plication of 10 Mmolof curcumin, chlorogenic acid, caffeic acid, ornithine decarboxylase activity in mouse epidermis was ex or ferulic acid together with 5 nmol of TPA twice a week for amined and the results are shown in Table 1. Curcumin strongly 19 weeks inhibited the number of tumors per mouse by 100, inhibited the TPA-induced increase in epidermal ornithine de 60,28, or 35%, respectively, and the percentage of animals with carboxylase activity in a dose-dependent manner. Topical ap tumors was inhibited 100, 31, 28, or 7%, respectively (Table plication of 0.5, 1, 3, or 10 ¿¿molofcurcumin with 5 nmol of 3). The topical application of 20 /¿molchlorogenic acid, 20 TPA inhibited the TPA-induced increases in epidermal orni /¿molcaffeic acid, or 50 jumol ferulic acid inhibited the average thine decarboxylase activity by 31,46, 84, or 98%, respectively. number of tumors per mouse by 91, 82, or 97%, respectively, Application of ira/w-retinoic acid (1.3 nmol), a known inhibitor and the percentage of animals with tumors was inhibited 61, of TPA-induced ornithine decarboxylase activity (21), inhibited 43, or 80%, respectively (Table 3). the TPA-induced increase in epidermal ornithine decarboxylase Effect of Curcumin on TPA- and Arachidonic Acid-induced activity by 88%. The effects of several naturally occurring Edema of Mouse Ears. The possibility that curcumin could phenolic acids that are structurally related to curcumin were inhibit TPA- and arachidonic acid-dependent inflammation was studied (Table 1, experiment 2). Topical application of 10 ¿tmol evaluated by studying the effects of curcumin on TPA- and of chlorogenic acid, caffeic acid, or ferulic acid inhibited TPA- arachidonic acid-induced edema of mouse ears. The application induced increases in ornithine decarboxylase activity by 25,42, of 0.4 or 1 /¿molof curcumin to an ear of the mouse 30 min or 46%, respectively, and higher doses of the phenolic acids before 0.5 nmol of TPA inhibited TPA-induced edema by 79 resulted in greater inhibition. The results of these studies indi or 97%, respectively (Table 4, experiment 1). In additional 5942
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Table I Effects ofcurcumin, chlorogenic acid, caffeic add, andferulic acid on TPA-induced ornithine decarboxylase activity in mouse epidermis TPA (S nmol) was applied topically alone or simultaneously with various doses of potential inhibitor in 200 »Iof acetone (experiment 1) or in 200 «Iof acetune:DMSO (90:10) (experiment 2). Mice were sacrificed 5 h later, the epidermis was isolated and ornithine decarboxylase activity was determined. Data are expressed as the mean ±SE of duplicate assays from at least six mice. decarboxylase inhibition of (pmol "CO2 released/ TPA-induced ornithine activity3146849888255542614662673991°ExperimentTreatment1 mgprotein/hr)31 decarboxylase AcetoneTPA ±12°3831 (5nmol)TPA ±6822659 /unni)TPA(5 nmol) + curcumin (0.5 ±8942082 pmol)TPA(5 nmol) + curcumin (1 ±544647 jimol)TPA(5 nmol) + curcumin (3 ±186°123 jimo!)TPA(5 nmol) + curcumin (10 ±54°472 nmol)2 (5 nmol) + retinoic acid (1.3 ±90°17±
Acetone:DMSO(90:10)TPA 11°2683 (5nmol)TPA ±2502022 ¿iinol)TPA(5 nmol) + chlorogenic acid (10 ±5091215± )TPA (5 nmol) + chlorogenic acid (20 //nini 118°1553 /imol)TPA(5 nmol) + caffeic acid (10 ±439°1068 ^mol)TPA(5 nmol) + caffeic acid (20 ±159°1470 nmol)TPA(5 nmol) + ferulic acid (10 ±190°1032 jimol)TPA(5 nmol) + ferulic acid (20 ±171°901 nmol)TPA(5 nmol) + ferulic acid (50 ±244°1636 jjmol)TPA(5 nmol) + curcumin (1 ±440°255 (5 nmol) + curcumin (10 mnol)Ornithine ±130°Percent Statistically different from TPA alone (P < 0.05).
Table 2 Effect ofcurcumin, chlorogenic acid, caffeic acid, andferulic acid on TPA-induced ['HJtHymidine incorporation into epidermal DNA TPA (2 nmol or 5 nmol) was applied topically alone or simultaneously with potential inhibitor in 200 j.1 of acetone (experiments 1 and 2) or in 200 it\ of acetone:DMSO (90:10) (experiment 3). Eighteen h after application of TPA alone or TPA with the various compounds, the mice were given injections of [3H]thymidine. Forty min later the mice were sacrificed, the epidermis was removed and radioactivity in epidermal DNA was determined. Data are expressed as the mean ±SE of 16 mice. incorporation inhibition into epidermal DNA of TPA-induced ExperimentTreatment1 (dpm/ngDNA)34.8 DNA synthesis4 Acetone ±5.5" TPA (2 nmol) 147.9 ±25.1 TPA (2 nmol) + curcumin (1 umuh 143.5 ±15.4 TPA (2 nmol) -I-curcumin (3 nmol) 132.0 ±17.5 14 TPA (2 nmol) + curcumin (10 nmol) 93.0 ±6.1° 49 nmol)2 TPA (2 nmol) + fluocinolone acetonide (2 50.5 ±10.5"33.4 8634
Acetone ±3.3° TPA (5 nmol) 136.7 ±9.5 TPA (5 nmol) + curcumin (1 nmol) 133.7± 15.6 TPA (5 nmol) + curcumin (3 nmol) 132.6 ±6.9 >3 TPA (5 nmol) + curcumin ( 10 ,,mol 106.7±4.0°27.7 297
Acetone:DMSO (90:10) ±2.9° TPA (5 nmol) 82.3 ±3.5 TPA (5 nmol) + chlorogenic acid (20 nmol) 78.3 ±4.6 TPA (5 nmol) + caffeic acid (20 nmol) 64.5 ±6.0°74.2 33 15' TPA (5 nmol) + ferulic acid (20 nmol)[3H]Thymidine ±7.4Percent Statistically different from TPA alone (P < 0.05). studies, the application of 3 or 10 ¿¿molofcurcumin to an ear inhibitors of TPA-induced tumor promotion and ornithine of the mouse 30 min before 1 /¿molofarachidonic acid inhibited decarboxylase activity in mouse skin, but these compounds are arachidonic acid-induced edema by 33 or 80%, respectively less effective than curcumin. Curcumin is widely used as a (Table 4, experiment 2). In another experiment, the topical yellow coloring agent and spice in commonly ingested foods, application of 1.5 or 5 ¿¿molofcurcumin30 min before 1 ^rnol and chlorogenic acid, caffeic acid, and ferulic acid are normally of arachidonic acid inhibited arachidonic acid-induced edema occurring constituents of many human foods and beverages. by 32 or 58%, respectively (Table 4, experiment 3). Phenidone Application of TPA to mouse skin results in the rapid accu and nordihydroguaiaretic acid are known inhibitors of arachi mulation of inflammatory cells such as neutrophils and mac donic acid metabolism (22, 23), and both of these compounds rophages (24) and an increase in the release of active oxygen were effective inhibitors of arachidonic acid-induced edema species (25, 26). As shown in Table 4, curcumin markedly (Table 4, experiments 2 and 3). inhibited TPA- and arachidonic acid-induced inflammation. The results could be explained by a possible effect of curcumin to inhibit the infiltration of inflammatory cells and/or to inhibit DISCUSSION the release and action of reactive oxygen species from these The results of the present study demonstrate that topical cells. application of curcumin inhibits TPA-induced epidermal orni It has been suggested that reactive oxygen species and other thine decarboxylase activity, epidermal DNA synthesis, and free radicals play an important role in tumor promotion (24- promotion of skin tumors in mice. The related compounds 27). Free radical-generating compounds such as benzoyl per chlorogenic acid, caffeic acid, and ferulic acid are also active as oxide, lauroyl peroxide, and chloroperbenzoic acid have tumor- 5943
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r A parallel the effects of these compounds as inhibitors of tumor promotion (see Ref. 13 and Table 3). Several inhibitors of arachidonic acid metabolism inhibit TPA-dependent tumor promotion in mouse skin (37, 38, 40, 41), and it is thought that products of arachidonic acid metab olism may play an important role in the tumor promotion process. Mouse epidermis has been reported to possess A5-, A8-, A12-,and A"-lipoxygenase activity (37, 42-44). HPETEs and HETEs, leukotrienes, and prostaglandins are among the arachidonic acid metabolic products that are believed to play a role in TPA-induced inflammation and tumor promotion (45- 47). It was found that TPA stimulates cell proliferation and increases the production of HETEs and prostaglandins in epi dermal cell culture (44). In addition, inhibition of arachidonic acid metabolism results in lowered levels of HETEs and pros taglandins in human platelets (48). These products of arachi donic acid metabolism may be important in cell proliferation
TPA(5nmol>-f since high levels of arachidonic acid, HPETEs, and HETEs are •^—' Curcumin(IO^mol) found in the hyperproliferative disease psoriasis (49). Fluoci- nolone acetonide and other glucocorticoids represent another 5 10 15 20 class of potent inhibitors of TPA-dependent tumor promotion WEEKS OF TPA APPLICATION (7), and these steroids are inhibitors of epidermal phospholipase Fig. 2. Effect of curcumin on TPA-induced tumor promotion in mouse skin. A; activity in vivo and inhibit the release of arachidonic acid Thirty mice/group were initiated with 200 nmol of DMBA. One week later the mice were treated topically with 5 nmol TPA alone or together with various doses from cellular membranes (50). Nordihydroguaiaretic acid and of curcumin in 200 ¡ilacetone twice a week for 20 weeks. A, tumors/mouse; B, quercetin are examples of As-lipoxygenase inhibitors that also percentage of mice with tumors. inhibit the TPA-dependent induction of ornithine decarboxyl ase activity and tumor promotion in mouse skin (23, 37, 41). promoting activity on mouse skin (27-29), and application of As indicated above, arachidonic acid metabolism is believed to TPA to mouse skin has been shown to stimulate the release give rise to reactive oxygen species and other free radicals, and and metabolism of arachidonic acid (30-33) and to increase the studies on the TPA-induced generation of reactive oxygen formation of reactive oxygen species (25, 26). Superoxide dis- species in mouse epidermal cells have indicated that inhibitors of arachidonic acid metabolism can diminish the chemilumi- mutase is an important enzymatic defense for the removal of Superoxide anión,and the biomimetic copper(II) (3,4-diisopro- nescense response that is associated with the generation of free radicals (51). It is possible that arachidonic acid metabolism pylsalicylic acid)2, which possesses Superoxide dismutase activ may provide a source of free radicals that play a role in tumor ity, has been demonstrated to inhibit TPA-induced ornithine promotion. The inhibitory effect of curcumin on arachidonic decarboxylase activity and tumor promotion in mouse skin (34, acid-dependent ear edema (Table 4) suggests that curcumin 35). Several inhibitors of TPA-dependent tumor promotion may block arachidonic acid metabolism by inhibiting epidermal possess antioxidant activity. Butylated hydroxytoluene (10), cyclooxygenase and/or lipoxygenase activity, or that curcumin butylated hydroxyanisole (10, 25), sodium selenite (36), quer- may serve as a scavenger of reactive free radicals which are cetin (37), nordihydroguaiaretic acid (38), a-tocopherol (39), produced during the metabolism of arachidonic acid. It is also ascorbic acid (12), and ascorbyl palmitate (12) are examples of possible that curcumin can interfere with the activity of epider compounds that possess antioxidant or reactive oxygen-scav mal phospholipase A2 and/or inhibit TPA-induced release of enging activity and that inhibit tumor promotion and/or certain arachidonic acid from membranes, since curcumin appears to biochemical events associated with tumor promotion in mouse be a more potent inhibitor of TPA-induced mouse ear edema skin. Although the antioxidant activity of chlorogenic acid has than arachidonic acid-induced ear edema (Table 4). Additional not been compared with that of curcumin, the relative antioxi studies are needed to determine the possible inhibitory effects dant activities of curcumin, caffeic acid, and ferulic acid roughly of curcumin, chlorogenic acid, caffeic acid, and ferulic acid on
Table 3 Effect of curcumin, chlorogenic acid, caffeic acid, and ferulic acid on TPA-induced tumor promotion in mouse skin Female mice were treated topically with 200 nmol of DMBA in 200 u\ of acetone. One week later the mice were treated topically with 5 nmol of TPA alone or together with curcumin, chlorogenic acid, caffeic acid, or ferulic acid in 200 pi of acetone:DMSO (90:10) twice a week for 19 weeks. The tumors per mouse are expressed as the mean ±SE from 30 mice. tumorsTreatmentAcetone:DMSO(90:10)TPA Percent of mice with wk048.323.36.724.711.534.50015wk060.740.06.743.323.044.83.3019wk067.946.626.746.738.563.513.3011wk0°3.79 wk0«5.53 wko-6.18
(5nmol)TPA ±1.370.91 ±1.842.50 ±1.842.50 (10nmol)TPA(5 nmol) + chlorogenic acid ±0.44°0.07 ±0.750.53 ±0.780.53
(20(imol)TPA(5 nmol) + chlorogenic acid ±0.05"3.47 ±0.10°4.43 ±0.05°4.43
^mol)TPA(5 nmol) + caffeic acid (10 ±0.090.31 ±1.531.12 ±1.891.12 /imol)TPA(5 nmol) + caffeic acid (20 ±0.21"2.21 +0.49°4.00 ±0.67°4.00 »imol)TPA(5 nmol) + ferulic acid (10 ±0.960°0°Tumors/mouse15±1.020.17 ±1.190.17 >imol)TPA(5 nmol) + ferulic acid (50 ±0.08°0°19 ±0.03°0° (5 nmol) + curcumin (10 nmol)11 " Statistically different from TPA alone (P < 0.05). 5944
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Table 4 Effect ofcurcumin on TPA- and arachidonic acid-induced edema of turmeric oleoresin. Recent studies have shown that feeding mouse ears In experiment 1, female CD-I mice (10 animals/group) were treated on the caffeic acid, chlorogenic acid, or ferulic acid (2% in the diet) to right ear with 25 nI of acetone or curcumin in acetone 30 min before the rats for 4 weeks resulted in hyperplasia throughout the fore- application of 25 n' of acetone or TPA in acetone. The mice were sacrificed 5 h stomach epithelium in the animals fed caffeic acid, but little or later by cervical dislocation, and ear punches from two animals were weighed together (6 mm diameter punch/animal). In experiments 2 and 3, female CD-I no effect was observed when the rats were fed chlorogenic acid mice (10 animals per group) were treated on the right ear with 25 jil of acetone or ferulic acid (57). or the potential inhibitor in acetone 30 min before the application of 25
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Mou-Tuan Huang, Robert C. Smart, Ching-Quo Wong, et al.
Cancer Res 1988;48:5941-5946.
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