The aim of this paper was to obtain low toxicity and high efficiency anti-tumor Chinese medicine through screening the combination ratios of Momordicae Semen and Epimedii Folium, and to explore the anti-tumor mechanism of the combination of two drugs by observing their effect on apoptosis-related proteins in cancer cells. Methyl thiazolyl tetrazolium(MTT) assay was used to observe the effect of drug combination on the proliferation of tumor cells from different tissue sources. The effects of the combination of the two drugs on tumor cells were analyzed by Compusyn software. Plate cloning assay was used to observe the effect of combination of these two drugs on the proliferation of A549 cells in vitro. The expression of reactive oxygen species(ROS) and apoptotic proteins p53, Bcl-2 and Bax were compared by using ROS kit and Western blot. Lewis lung cancer model was used to observe the anti-tumor effect of drugs in vivo. The results showed that the anti-tumor effect of their ethanol extract was more significant than that of water extract, and the anti-proliferation effect was strongest when the ratio was 1∶1(P<0.05). Compusyn analysis showed that the combination of the two drugs had synergistic effect. Further studies showed that after combined use, the number of clonogen formation in A549 cells was significantly reduced(P<0.01); ROS production was increased; the expression of apoptosis-related protein p53 was up-regulated, and the ratio of Bcl-2/Bax was decreased. In vivo animal study showed that the tumor inhibition rate was 53.06%(P<0.05) in the high dose group. As compared with the single use of the two drugs, the combination of the two drugs had more significant anti-proliferative effect on tumors, and the optimum ratio was 1∶1. The combination of the two drugs at a ratio of 1∶1 inhibited the proliferation of various tumor cells, and had no significant effect on normal liver cells LO2 when compared with other ratios. Therefore, it can be preliminarily inferred that the combination of the two drugs may have the effect of synergism and detoxification. Further studies showed that the combination of the two drugs can significantly inhibit the proliferation of A549 cells, and its mechanism may be related to the activation of endogenous apoptotic pathway. In vivo experiments also showed that the tumor inhibition rate increased with the increase of drug concentration.
A549 Cells ; Animals ; Antineoplastic Agents, Phytogenic/pharmacology* ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Drugs, Chinese Herbal/pharmacology* ; Epimedium/chemistry* ; Humans ; Lung Neoplasms/drug therapy* ; Momordica/chemistry* ; Neoplasms, Experimental/drug therapy* ; Plant Leaves/chemistry*

This study explored how bitter melon powder (BMP) alters the colonic microenvironment during the development of obesity-associated fatty liver in rats. We observed that BMP effectively inhibited the body weight gain and lipid accumulation in the liver, ameliorated glucose intolerance, and increased the colon weight after an 8-week treatment compared to that in the high-fat diet (HFD) group. BMP significantly decreased fecal water toxicity towards HT-29 cells, as revealed by the cell counting kit (CCK)-8 assay results, and the mRNA expression of Toll-like receptor 4 (TLR4) in colon mucosa. Additionally, gut permeability in the BMP group was restored to normal levels. Finally, BMP alleviated the inflammatory state of the rat colon mucosa and liver tissues as well as the systemic inflammation.
Animals ; Colon ; drug effects ; Dietary Fats ; administration & dosage ; adverse effects ; Fatty Liver ; etiology ; prevention & control ; Feces ; chemistry ; HT29 Cells ; Humans ; Momordica charantia ; Obesity ; complications ; Powders ; Rats

Nature is a rich source of medicinal plants and their products that are useful for treatment of various diseases and disorders. Momordica charantia, commonly known as bitter melon or bitter gourd, is one of such plants known for its biological activities used in traditional system of medicines. This plant is cultivated in all over the world, including tropical areas of Asia, Amazon, east Africa, and the Caribbean and used as a vegetable as well as folk medicine. All parts of the plant, including the fruit, are commonly consumed and cooked with different vegetables, stir-fried, stuffed or used in small quantities in soups or beans to give a slightly bitter flavor and taste. The plant is reported to possess anti-oxidant, anti-inflammatory, anti-cancer, anti-diabetic, anti-bacterial, anti-obesity, and immunomodulatory activities. The plant extract inhibits cancer cell growth by inducing apoptosis, cell cycle arrest, autophagy and inhibiting cancer stem cells. The plant is rich in bioactive chemical constituents like cucurbitane type triterpenoids, triterpene glycosides, phenolic acids, flavonoids, essential oils, saponins, fatty acids, and proteins. Some of the isolated compounds (Kuguacin J, Karaviloside XI, Kuguaglycoside C, Momordicoside Q-U, Charantin, α-eleostearic acid) and proteins (α-Momorcharin, RNase MC2, MAP30) possess potent biological activity. In the present review, we are summarizing the anti-oxidant, anti-inflammatory, and anti-cancer activities of Momordica charantia along with a short account of important chemical constituents, providing a basis for establishing detail biological activities of the plant and developing novel drug molecules based on the active chemical constituents.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; Humans ; Momordica charantia ; chemistry ; Neoplasms ; drug therapy ; Plant Extracts ; chemistry ; pharmacology

BACKGROUND/AIMS: The major compounds of Cochinchina momordica seed extract (SK-MS10) include momordica saponins. We report that the gastroprotective effect of SK-MS10 in an ethanol-induced gastric damage rat model is mediated by suppressing proinflammatory cytokines and downregulating cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase (5-LOX), and the activation of calcitonin gene-related peptide. In this study, we evaluated the gastroprotective effects of SK-MS10 in the nonsteroidal anti-inflammatory drug (NSAID)-induced gastric damage rat model. METHODS: The pretreatment effect of SK-MS10 was evaluated in the NSAID-induced gastric damage rat model using aspirin, indomethacin, and diclofenac in 7-week-old rats. Gastric damage was evaluated based on the gross ulcer index by gastroenterologists, and the damage area (%) was measured using the MetaMorph 7.0 video image analysis system. Myeloperoxidase (MPO) was measured by enzyme-linked immunosorbent assay, and Western blotting was used to analyze the levels of cyclooxygenase (COX)-1, COX-2, cPLA2, and 5-LOX. RESULTS: All NSAIDs induced gastric damage based on the gross ulcer index and damage area (p<0.05). Gastric damage was significantly attenuated by SK-MS10 pretreatment compared with NSAID treatment alone (p<0.05). The SK-MS10 pretreatment group exhibited lower MPO levels than the diclofenac group. The expression of cPLA2 and 5-LOX was decreased by SK-MS10 pretreatment in each of the three NSAID treatment groups. CONCLUSIONS: SK-MS10 exhibited a gastroprotective effect against NSAID-induced acute gastric damage in rats. However, its protective mechanism may be different across the three types of NSAID-induced gastric damage models in rats.
Animals ; Anti-Inflammatory Agents, Non-Steroidal/adverse effects ; Arachidonate 5-Lipoxygenase/drug effects ; Calcitonin Gene-Related Peptide/drug effects ; Cyclooxygenase 1/drug effects ; Cyclooxygenase 2/drug effects ; Disease Models, Animal ; Gastric Mucosa/chemistry/drug effects ; Group IV Phospholipases A2/drug effects ; Male ; Momordica/*chemistry ; Peroxidase/drug effects ; Plant Extracts/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Seeds/*chemistry ; Stomach Ulcer/chemically induced/*prevention & control ; Treatment Outcome

OBJECTIVETo develop an HPLC for determination of aglycone of momordicoside L in Momordica charantia.

METHODA Kromasil C18 (4.6 mm x 150 mm, 5 microm) column was used. The mobile phase was acetonitrile-H2O (64:36), the flow rate was 1.0 mL x min(-1) and the UV detection wavelength was set 203 nm.

RESULTThe calibration curves were linear from of 0.025 microg to 1 microg (r =0.9911), the contents of aglycone of momordicoside L in Shandong, Henan, Hebei, Jiangxi are 0.211, 0.033, 0.013, 0.007 mg x g(-1), respectively.

CONCLUSIONThe method is simple and reliable for determination of aglycone of momordicoside L in M. charantia.

Calibration ; Chromatography, High Pressure Liquid ; methods ; Momordica charantia ; chemistry ; Saponins ; analysis

OBJECTIVETo clone and produce ribosome inactivating protein MAP30 from the seeds of Momordica charantia L(bitter melon), and to evaluate the biological activity of the recombinant protein.

METHODSThe DNA sequence encoding MAP30 was cloned from the fresh seeds of Momordica charantia by PCR, the target DNA fragments were sequenced after T-A cloning. The expression plasmid was constructed by inserting the MAP30 fragment into vector pET30a. MAP30 was expressed in E.coli by addition of IPTG into final concentration of 1.0 mmol/L. The recombinant MAP30 was identified by SDS-PAGE, and the biological activity of MAP30 protein was evaluated by using MTT assay in cancer cells and normal cells following fluid-phase endocytosis.

RESULTThe nucleotide and amino acid sequences of the cloned MAP30 were identical with those of reported MAP30. The solubility of recombinant protein was analyzed by SDS-PAGE, and the MAP30 was mainly produced in soluble form. The recombinant MAP30 showed a greater cytotoxicity to cancer cells than that to normal cells.

CONCLUSIONThe gene of MAP30 has been successfully cloned.The recombinant MAP30 protein expressed by E.coli is bioactive.

Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Momordica charantia ; chemistry ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Ribosome Inactivating Proteins, Type 2 ; biosynthesis ; genetics ; metabolism ; Seeds ; chemistry ; Transformation, Bacterial

Luo han kuo fruit (Siraitia grosvenori Swingle), a fruit native to China, has been used as a natural sweetening agent for centuries and has been reported to be beneficial for diabetic population. However, limited research has been conducted to elucidate the relationship between the sweetening action and biological parameters that may be related to potential health benefits of LHK fruit (Luo Han Kuo fruit). The present study examined the effect of LHK fruit and its chemical components on insulin secretion using an in vitro cell model system. Mogroside V is the most abundant and the sweetest chemical component among the mogrosides in LHK fruit. The experimental data demonstrated that the crude LHK extract stimulated the secretion of insulin in pancreatic beta cells; furthermore, pure mogroside V isolated from LHK fruit also exhibited a significant activity in stimulating insulin secretion by the beta cells, which could partially be responsible for the insulin secretion activity of LHK fruit and fruit extract. The current study supports that LHK fruit/extract has the potential to be natural sweetener with a low glycemic index, and that mogroside V, possible other related mogrosides, can provide a positive health impact on stimulating insulin secretion.
Animals ; Cucurbitaceae ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fruit ; chemistry ; Insulin ; secretion ; Insulin-Secreting Cells ; secretion ; Momordica ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Sweetening Agents ; isolation & purification ; pharmacology ; Triterpenes ; isolation & purification ; pharmacology

Momordica charantia L. is a vegetable widely cultivated around the world. Its fruits have been used in Asian countries as a folk medicine for the treatment of non-insulin-dependent diabetes mellitus. Here we report eight compounds isolated from the fruits of M. charantia. On the basis of NMR and MS spectroscopic analyses, these compounds were identified as momordicolide ((10E)-3-hydroxyl-dodeca-10-en-9-olide, 1), monordicophenoide A (4-hydroxyl-benzoic acid 4-O-beta-D-apiofuranosyl (1 --> 2)-O-beta-D-glucopyranoside, 2), dihydrophaseic acid 3-O-beta-D-glucopyranoside (3), 6,9-dihydroxy-megastigman-4,7-dien-3-one (blumenol, 4), guanosine (5), adenosine (6), uracil (7) and cytosine (8). Among them, 1 and 2 are new compounds. Compounds 3-5 were isolated from this plant for the first time.
Fruit ; chemistry ; Glycosides ; analysis ; chemistry ; Momordica charantia ; chemistry ; Triterpenes ; analysis ; chemistry

In this study, polyacrylicacid precipitation alkalescence protein from Momordica charantia L. seeds was studied, and the effect of conditions on experiment was also evaluated. Isoelectric precipitation is achieved by adjusting the pH of a protein solution and is based on that a protein's solubility is at minimum at its pI. The sample was titrated to pH 6.0 with citric acid, and 14.62% proteins were precipitated. With hydrochloric acid to pH 4.0, 32.49% proteins were precipitated. With the acetic acid to pH 6.0 and pH 4.0, 26.17% and 38.72% proteins were precipitated, respectively. In the 1 mL Bitter melon seeds extraction(pH 4.0) adjusted by acetic acid, hydrochloric acid and citric acid, the optimum dosage of PAA (1%) precipiting alkalescency protein (pl 8.65-9.30) was 100 microL, 120 microL and 100 microL, respectively. The respective extraction (1mL) was titrated to pH 5.0, pH 4.0, and pH 3.0 by acetic acid. After isoelectric precipitation, the PAA precipitation protein was performed. When concentration of PAA (1%) was 160 microL/mL, the protein decreased in the supernatant was 33.77% at pH 5.0, and 43.56% at pH 3.0. When concentration of PAA (1%) was 120 microL/mL, the protein decreased in the supernatant was 30.83% at pH 4.0. PAA-Protein complex could redissolve in alkaline conditions (pH > 9.0) and the protein most easilly redissolved when the NaCL was 3.0%. The bitter melon seeds extraction after PAA purification flowed through the Sephadex G-75 columns. The peaks I and II were obtained after 175 min and 300 min, respectively. SDS-PAGE and IEF analysis showed that the molecule weight from peaks I was 30 kD with pI 9.5, peaks II 10 kD with pI 9.3.
Acrylic Resins ; chemistry ; Chemical Precipitation ; Momordica charantia ; chemistry ; Plant Extracts ; chemistry ; Plant Proteins ; chemistry ; isolation & purification ; Seeds ; chemistry

OBJECTIVETo establish a weight-marking method and evaluate Siraitia grosvenorii germplasms.

METHODThe characters of 21 kinds of S. grosvenorii germplasms in the field and the lab were analyzed, seven important characters were selected to weight the marks. A general evaluation index was made and used to evaluate S. grosvenorii germplasms.

RESULTThe evaluation result of 21 kinds of S. grosvenorii germplasms by the weight-marking method was consistent with production practice. Meanwhile, the new variety Yongqing No. 1 and major cultivars were the superior germplasms.

CONCLUSIONA rational technique system of evaluating S. grosvenorii germplasms was established, and the superior germplasms were selected.

Biomass ; China ; Drugs, Chinese Herbal ; analysis ; chemistry ; standards ; Fruit ; anatomy & histology ; chemistry ; Glucosides ; analysis ; Momordica ; anatomy & histology ; chemistry ; growth & development ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development ; Quality Control ; Triterpenes ; analysis