We examined the inhibitory effects of loquat methanol extract on the adhesion, migration, invasion and matrix metalloproteinase (MMP) activities of MDA-MB-231 human breast cancer cell line. Cells were cultured with DMSO or with 10, 25, or 50 microg/ml of loquat methanol extract. Both leaf and seed extracts significantly inhibited growth of MDA-MB-231 cells in a dose-dependent manner, although leaf extract was more effective. Adhesion and migration were significantly inhibited by loquat extracts in a dose-dependent manner. Loquat extract also inhibited the invasion of breast cancer cells in a dose-dependent manner and leaf extract was more effective than seed extract. MMP-2 and MMP-9 activities were also inhibited by loquat extract. Our results indicate that methanol extracts of loquat inhibit the adhesion, migration and invasion of human breast cancer cells partially through the inhibition of MMP activity and leaf extract has more anti-metastatic effects in cell based assay than seed extract. Clinical application of loquat extract as a potent chemopreventive agent may be helpful in limiting breast cancer invasion and metastasis.
Breast Neoplasms ; Cell Line ; Dimethyl Sulfoxide ; Eriobotrya ; Humans ; Methanol ; Neoplasm Metastasis ; Seeds

OBJECTIVETo find out the change regularity of triterpenoid acids content in leaves of Eriobotrya japonica. (Folium Eriobotryae) in different seasons.

METHODThe content of triterpenoid acids was determined by RP-HPLC.

RESULTThe content changes of oleanolic acid, ursolic acid, corosolic acid were minor in the different seasons. The content of oleanolic acid and ursolic acid were highest in April while the content of corosolic acid was highest in July. The content of maslinic acid has a significant change, the material which collected in November has the highest content in a year.

CONCLUSIONThe content of triterpenoid acids changes regularly.

Drugs, Chinese Herbal ; analysis ; Eriobotrya ; chemistry ; Plant Leaves ; chemistry ; Seasons ; Triterpenes ; analysis

OBJECTIVETo extract corosolic acid from loquat leaves for medical use.

METHODSLoquat leaves were boiled in water to remove the water-soluble substances followed by 3 cycles of extraction with 25% aqueous methanol for 30 min and then by 95% aqueous methanol for 1 h at 80 degrees celsius;. After cooling at room temperature and filtration, the extract was treated with activated carbon to remove chlorophyll, and the liquid was filtered and concentrated to allow precipitation. The sediment was washed to obtain the total crude triterpene acid, which was further dissolved with methanol and purified by high-performance liquid chromatography (HPLC). The fractions including corosolic acid were collected, concentrated with vacuum distillation, and dried to obtain corosolic acid product, which was analyzed with HPLC.

RESULTSHPLC analysis of the extracts showed that the percentages of corosolic acid were 4.66%, 2.42%, and 24.18% in crude corosolic acid extracted with methanol, boiling water, and 95% aqueous methanol, respectively. After purification with HPLC, the purity of corosolic acid in the product exceeded over 80%.

CONCLUSIONThe optimal extraction method, which is convenient and cost-effective, is established for extracting corosolic acid from loquat leaves for medical use.

Chromatography, High Pressure Liquid ; methods ; Eriobotrya ; chemistry ; Plant Leaves ; chemistry ; Technology, Pharmaceutical ; Triterpenes ; isolation & purification

BACKGROUND/OBJECTIVES: The present study was conducted to examine the inhibitory effect of loquat leaves on MDA-MB-231 cell proliferation and invasion. MATERIALS/METHODS: Female athymic nude mice were given a subcutaneous (s.c.) inoculation of MDA-MB-231 cells and randomly grouped to receive a s.c. injection of either 500 mg/kg ethanol, water extract or vehicle five times a week. Tumor growth, mitotic rate and necrosis were examined. MDA-MB-231 cells were cultured with DMSO or with various concentrations of loquat water or ethanol extract. Proliferation, adhesion, migration, invasion and matrix metalloproteinase (MMP) activity were examined. RESULTS: Tumor growth of xenograft nude mouse was significantly reduced by loquat extracts. The results of mitotic examination revealed that loquat extracts reduced tumor cell division. Both ethanol and water extracts significantly inhibited MDA-MB-231 cell proliferation. The protein expression of ErbB3 was significantly down-regulated by loquat leaf extracts. Loquat leaf extracts increased apoptosis of MDA-MB-231 cells following 24 hour incubation and the ethanol extract was more potent in inducing apoptosis than the water extract. Furthermore, loquat extracts inhibited adhesion, migration and invasion of MDA-MB-231 cells. MMP activity was significantly inhibited by loquat extracts. CONCLUSION: Our results show that extracts of loquat inhibit the growth of tumor in MDA-MB-231 xenograft nude mice and the invasion of human breast cancer cells, indicating the inhibition of tumor cell proliferation and invasion.
Animals ; Apoptosis ; Breast Neoplasms ; Cell Division ; Cell Proliferation ; Dimethyl Sulfoxide ; Eriobotrya* ; Ethanol ; Female ; Heterografts* ; Humans ; Mice ; Mice, Nude* ; Necrosis ; Water

Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
Biomass ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Eriobotrya ; chemistry ; growth & development ; metabolism ; Triterpenes ; analysis ; metabolism

The near infrared (NIR) spectroscopy technique has been applied in many fields because of its advantages of simple preparation, fast response, and non-destructiveness. We investigated the potential of NIR spectroscopy in diffuse reflectance mode for determining the soluble solid content (SSC) and acidity (pH) of intact loquats. Two cultivars of loquats (Dahongpao and Jiajiaozhong) harvested from two orchards (Tangxi and Chun'an, Zhejiang, China) were used for the measurement of NIR spectra between 800 and 2500 nm. A total of 400 loquats (100 samples of each cultivar from each orchard) were used in this study. Relationships between NIR spectra and SSC and acidity of loquats were evaluated using partial least square (PLS) method. Spectra preprocessing options included the first and second derivatives, multiple scatter correction (MSC), and the standard normal variate (SNV). Three separate spectral windows identified as full NIR (800approximately2500 nm), short NIR (800approximately1100 nm), and long NIR (1100approximately2500 nm) were studied in factorial combination with the preprocessing options. The models gave relatively good predictions of the SSC of loquats, with root mean square error of prediction (RMSEP) values of 1.21, 1.00, 0.965, and 1.16 degrees Brix for Tangxi-Dahongpao, Tangxi-Jiajiaozhong, Chun'an-Dahongpao, and Chun'an-Jiajiaozhong, respectively. The acidity prediction was not satisfactory, with the RMSEP of 0.382, 0.194, 0.388, and 0.361 for the above four loquats, respectively. The results indicate that NIR diffuse reflectance spectroscopy can be used to predict the SSC and acidity of loquat fruit.
Eriobotrya ; chemistry ; Hydrogen-Ion Concentration ; Least-Squares Analysis ; Spectroscopy, Fourier Transform Infrared ; methods ; Spectroscopy, Near-Infrared ; methods

A total of twelve compounds were isolated from the ethyl acetate of the water extract of honey-fried Eriobotrya japonica through column chromatography over silica gel,Sephadex LH-20,RP-18,and preparative HPLC. Their structures were established by MS,1 D NMR and 2 D NMR data as japonicanoside A( 1),nerolidol-3-O-α-L-rhamnopyranosyl-( 1→2)-β-D-glucopyranoside( 2),nerolidol-3-O-α-L-rhamnopyranosyl-( l→4)-α-L-rhamnopyranosyl-( 1 → 2)-[α-L-( 4-trans-feruloyl)-rhamnopyranosyl-( 1 → 6) ]-β-D-glucopyranoside( 3),( +)-catechin( 4),(-)-epicatechin( 5),kaempferol 3-O-α-L-rhamnopyranoside( 6),quercitrin( 7),quercetin-3-O-β-D-galactopyranoside( 8),quercetin-3-O-β-glucopyranoside( 9),vanillin( 10),protocatechuic aldehyde( 11),and maltol( 12). Among them,1 is a new phenolic glycoside.
Chromatography, High Pressure Liquid ; Eriobotrya ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Honey ; Magnetic Resonance Spectroscopy ; Phytochemicals ; chemistry ; isolation & purification

BACKGROUND: Although Eriobotrya japonica leaves have been studied as a raw material for various cosmetic products, little is known about the anti-oxidant, anti-inflammatory, and anti-melanogenic activities of Eriobotrya japonica leaf ethanol extract (EJEE). METHODS: This study was conducted to evaluate the anti-oxidant, anti-inflammatory, and anti-melanogenic activities of EJEE using different in vitro models. In addition, we investigated the potential irritation of EJEE to skin and eye using animal alternative tests. RESULTS: The total content of polyphenols, one of the active constituents of EJEE, was analyzed by high-performance liquid chromatography and found to contain 88.68 mg tannic acid equivalent/g. EJEE showed a concentration-dependent 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical scavenging activity, and a superoxide dismutase-like activity. The anti-inflammatory effect of 0.5% (w/v) EJEE was demonstrated by a reduction in lipopolysaccharide-induced nitric oxide and tumor necrosis factor-alpha levels in RAW 264.7 cells. EJEE also significantly inhibited melanogenesis in melanocyte stimulating hormone-induced B16F1 cells. EJEE did not show any irritation in skin and eye in animal alternative test. CONCLUSIONS: These results indicate that the EJEE possesses anti-oxidant, anti-inflammatory, and anti-melanogenic activities, while it did not induce toxicity or irritation in neither skin nor eye. Therefore, EJEE can be used as a cosmetic ingredient for skin improvement.
Animals ; Chromatography, Liquid ; Eriobotrya ; Ethanol ; In Vitro Techniques ; Melanocytes ; Nitric Oxide ; Polyphenols ; RAW 264.7 Cells ; Skin ; Superoxides ; Tannins ; Tumor Necrosis Factor-alpha

The loquat is widely cultivated in China, its succulent fruits, leaves and flower are used as a traditional medicine for the treatment of many diseases. The study is aimed to analyse the content of the four triterpene compounds ( ursolic acid, corosolic acid, maslinic acid, oleanolic acid) in different organs, and investigate the dynamic changes in different phenological period. The triterpenic acids content in the samples was measured by HPLC based on the plant phenological observations. The results showed that order of four triterpenic acids content in different organs from high to low was defoliation (23.2 mg x g(-1)) > mature leaves (21.7 mg x g(-1)) > young leaves (17.5 mg x g(-1)) > fruits (7.36 mg x g(-1)) > flowers (6.40 mg x g(-1)). The triterpenic acids were not detected in the seeds. The total amount of the four triterpenic acids in the loquat leaves collected in the different phenological stages of sprout, flower bud, blossom and fruit varied between 17.8 and 26.2 mg x g(-1) (defoliation), 16.5 and 23.5 mg x g(-1) (mature leaves), 14.7 and 21.5 mg x g(-1) (young leaves), respectively. The content increased progressively with the leaf development, maturation and aging. There was a higher level of the dry material and triterpenic acids accumulation in the mature leaves during fruit enlargement. This paper attempts to present the case for medicinal plants of a broad geographical distribution to study on the secondary metabolites and harvesting time.
China ; Chromatography, High Pressure Liquid ; Eriobotrya ; chemistry ; growth & development ; Flowers ; chemistry ; growth & development ; Fruit ; chemistry ; growth & development ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; growth & development ; Triterpenes ; analysis

OBJECTIVETo establish a method for the determination of the papaverine content in Qiangli Pibalu by HPLC.

METHODA C18 column with a solvent system of acetonitrile-0.02 mol x L(-1) sodium dihydrogen phosphate (0.2% triethylamine, phosphoric acid, at pH 3) (25:75) and UV detection 240 nm were used. The flow rate was 1.0 mL x min(-1). The column temperature was maintained at 40 degrees C.

RESULTThere was a good linear relationship between the absorption value and the concentration in the range of 0.020 2-0.100 5 microg for papaverine. The average recovery rates were 99.1% (RSD 2.3%).

CONCLUSIONThe method is simple, accurate and can be used to determine the contents in Qiangli Pibalu.

Antitussive Agents ; chemistry ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Eriobotrya ; chemistry ; Expectorants ; chemistry ; pharmacology ; Papaver ; chemistry ; Papaverine ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results