Determination and effect of five alkaloids from extracts of Piper longum on rats with experimental myocardial ischemia induced by injection of pituitrin / 药学实践杂志

Objective To determine the content of five alkaloids from extracts of Piper longum and test the pharmacodynamic effect of them on rats with experimental myocardial ischemia induced by injection of pituitrin. Methods The content of five alkaloids was determined simultaneously by HPLC. The experimental myocardial ischemia in rats was induced by injection of pituitrin, and the absolute value of T wave change and change of heart rate before and after model establishment were chosen to be the observation index. The effects of large, medium and small dose groups were evaluated. Results Three batches of samples were analyzed, with the contents of piperine for 56.1%, 49.7%, 51.6%; N-isobutyl-(2E,4E)-octadecatrienamide for 4.5%, 4.2%, 4.3%; guineensine for 0.46%, 0.38%, 0.40%; piplartine for 1.73%, 1.67%, 1.70% and piperamide for 0.55%, 0.46%, 0.49%, respectively. All dose groups from extracts of piper longum had significantly reduced the absolute value of T wave and almost have no effect on the change of heart rate, except the high dose group showed the effect of reducing heart rate at some time . Conclusion The HPLC method was suitable for the simultaneous determination of five alkaloids from extracts of Piper longum. It was shown that extracts of Piper longum had good bioactivity in anti-myocardial ischemia.

Piperlongumine suppressed osteoclastogenesis in RAW264.7 macrophages

Piperlongumine (PL) is a natural product found in long pepper (Piper longum). The pharmacological effects of PL are well known, and it has been used for pain, hepatoprotection, and asthma in Oriental medicine. No studies have examined the effects of PL on bone tissue or bone-related diseases, including osteoporosis. The current study investigated for the first time the inhibitory effects of PL on osteoclast differentiation, bone resorption, and osteoclastogenesis-related factors in RAW264.7 macrophages stimulated by the receptor activator for nuclear factor-κB ligand (RANKL). Cytotoxicity was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and osteoclast differentiation and bone resorption were confirmed by tartrate-resistant acid phosphatase (TRAP) staining and pit formation analysis. Osteoclast differentiation factors were confirmed by western blotting. PL exhibited toxicity in RAW264.7 macrophages, inhibiting osteoclast formation and bone resorption, in addition to inhibiting the expression of osteoclastogenesis-related factors, such as tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Fos, and NFATc1, in RANKL-stimulated RAW264.7 macrophages. These findings suggest that PL is suitable for the treatment of osteoporosis, and it serves as a potential therapeutic agent for various bone diseases.

Research progress and application of Piper longum in traditional Chinese medicine and traditional Mongolian medicine / 中草药

Piper longum is the immature ear of Pepper family of Papper. Although P. longum is used to treat gastrointestinal diseases in traditional Chinese and Mongolian medicines, there are significant differences in compatibility and treatment of diseases. In this paper, the traditional Mongolian medicine P. longum in Chinese usage, taste, function, and clinical compatibility research were analyzed and compared. Through literature research and statistical analysis, we would sort out the records of P. longum in Chinese materia medica and Mongolian materia medica, combined with computer software analysis, analyze the frequency of use in clinical medicine P. longum and drug law; This review introduced the progress of research on chemical constituents and pharmacological activities of P. longum compatibility as well as in clinical medicine in the Chinese, hoping to provide the basis for the application of P. longum in the field of traditional Chinese and Mongolian medicines.

Identify metabolites of Piper longum alkaloids in feces and urine of rat by HPLC-FT-MS / 中国中药杂志

The metabolites were detected in feces and urine of rats orally administrated alkaloids of Piper longum by using high performance liquid chromatography coupled with a Fourier Transform ion cyclotron resonance mass spectrometer (HPLC-FT-MS). According to the mass spectrometric data and reported literature, the structures of metabolites were identified. Several metabolites were analyzed and belonged to piperine, piperanine, piperlonguminine, Δα,β-dihydropiperlonguminine and pellitorine, respectively. The metabolites of alkaloids from P. longum alkaloids were produced through Ⅰ phase and Ⅱ phase metabolism reaction, and were excreted with urination and defecation. The approach provided a rapid method for characterizing the metabolites of P. longum alkaloids and gave the truly active structures and the action mechanism of their neuroprotective effects.

Comparative metabolism of three amide alkaloids from Piper longum in five different species of liver microsomes / 中国中药杂志

Piperine, piperlonguminine and pellitorine are three major amide alkaloids from Piper longum, showing a variety of pharmacological activities. In order to investigate the different metabolism pathways of these compounds in five species of liver microsomes in vitro, the data of full mass spectrum, and MS2, MS3 spectra of these three alkaloids were collected and analyzed by using ultra-high-performance liquid chromatography coupled with a LTQ-orbitrap mass spectrometer (UHPLC-LTQ-Orbitrap MS); gragment ion information was collected and combined with fragmentation regularities of mass spectra and accurate mass spectrometry data of metabolites, to compare the metabolism difference of three amide alkaloids in liver microsomes of human, rhesus monkey, Beagle dogs, rats and mice. 3 metabolites of piperine, 2 metabolites of piperlonguminine and 1 metabolite of pellitorine were identified quickly. The results showed that the major metabolic pathways of these amide alkaloids in liver microsomes were methylenedioxy group demethylation and oxidation reaction, and metabolic rates were different between species. This study provides basis for further research on in vivo metabolism of piperine analogues from Piper longum.

Cell cycle inhibitory activity of Piper longum against A549 cell line and its protective effect against metal-induced toxicity in rats.

Anticancer potential of Piper longum fruit against human cancer cell lines (DU-145 prostate, A549 lung, THP-1 leukemia, IGR-OVI-1 ovary and MCF-7 breast) as well as its in vitro and in vivo biochemical efficacy in AlCl3-induced hepatotoxicity were evaluated in the rats. Dried samples were extracted with several solvents using soxhlet apparatus. Flavonoid content in chloroform, benzene, ethyl alcohol and aqueous extracts of fruit was 19, 14, 12 and 11 μg quercetin equivalent/mg of sample, respectively. Hexane extracts exhibited 90-92% cytotoxicity against most of the test cell lines (A549, THP-1, IGR-OVI-1 and MCF-7), while benzene extract displayed 84-87% cytotoxicity against MCF-7, IGR-OV-1 and THP-1 cell lines. Among extracts, hexane, benzene and acetone extracts demonstrated considerable cytotoxicity (91-95%) against A549 (lung cancer) cell line in Sulforhodamine B dye (SRB) assay. Cell cycle analysis revealed that hexane, benzene and acetone extracts produced 41, 63 and 43% sub-G1 DNA fraction, demonstrating cell cycle inhibitory potential of these extracts against A549 cell line. Chloroform, ethyl alcohol and aqueous extracts displayed 71, 64 and 65% membrane protective activity, respectively in lipid peroxidation inhibition assay. P. longum fruit extracts also ameliorated AlCl3-induced hepatotoxicity, as indicated by alterations observed in serum enzymes ALP, SGOT and SGPT activity, as well as creatinine and bilirubin contents. In conclusion, study established the cytotoxic and hepatoprotective activity in P. longum extracts.

Protective Effect of Hexane and Ethanol Extract of Piper Longum L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures

OBJECTIVES: Gentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine in Asia and Pacific islands, which has been reported to exhibit a wide spectrum of activity, including antioxidant activity. In this study, we evaluated the effect of hexane:ethanol (2:8) PL extract (subfraction of PL [SPL] extract) on GM-induced hair cell loss in basal, middle and apical regions in a neonatal cochlea cultures. METHODS: The protective effects of SPL extract were measured by phalloidin staining of cultures from postnatal day 2-3 mice with GM-induced hair cell loss. The anti-apoptosis activity of SPL extract was measured using double labeling by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and myosin-7a staining. The radical-scavenging activity of SPL extract was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. RESULTS: SPL extract at a concentration of 1 microg/mL significantly inhibited GM-induced hair cell loss at basal and middle region of cochlea, while 5 microg/mL was effective against apical region hair cell loss. The protective effect of SPL extract was concentration dependent and hair cells retained their stereocilia in explants treated with SPL extract prior to treatment with 0.3 mM GM. SPL extract decreased GM-induced apoptosis of hair cells as assessed by TUNEL staining. The outer hair and inner hair counts were not decreased in SPL extract treated groups in compare to GM treated explants. Additionally, SPL extract showed concentration dependent radical scavenging activity in a DPPH assay. CONCLUSION: An anti-apoptosis effect and potent radical scavenger activity of SPL extract protects from GM-induced hair cell loss at basal, middle and apical regions in neonatal cochlea cultures.

Effect of Pepper and Piper Longum on Hepatic Energy Metabolism Factors in Rats / 世界科学技术-中医药现代化

This study was aimed to explore the effect of heat-nature Chinese herbs of Pepper and Piper longum on hepatic energy metabolism of rats in order to research the impact of ratˊs physical activities with heat-nature Chinese herbs and establish the research foundation of Chinese herbal medicine nature . A total of 30 rats were randomly divided into three groups . Saline was given in the control group . Pepper water extract was given in the Pepper group . And Piper longum water extract was given in the Piper longum group . The administration dosage was 10 ml/kg intragastrically for 30 days . Changes of hepatic energy metabolism related enzymes were determined . The results showed that compared with the control group , Ca2+-ATPase , Ca2+-Mg2+-ATPase , and LDH enzyme of the Pepper group and Piper longum group were significantly increased . And the hepatic glycogen content was significantly reduced . It can obviously increase energy intake , digestible energy and metabolizable energy per body weight in rats . There was no statistical significance between the Pepper group and Piper longum group . It was concluded that the heat-nature Chinese herbs of Pepper and Piper longum can promote the decomposition of hepatic glycogen , increase the activity of LDH enzyme , Ca2+-ATPase , Ca2+-Mg2+-ATPase . It increases the consumption of ATPase in order to increase energy intake , digestible ener-gy and metabolizable energy per body weight in rats and promote the hepatic energy metabolism .

Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation

Piperine and piperlongumine, alkaloids having diverse biological activities, commonly occur in roots of Piper longum L., Piperaceae, which have high commercial, economical and medicinal value. In present study, rapid, validated HPTLC method has been established for the determination of piperine and piperlongumine in methanolic root extract and its commercial formulation 'Mahasudarshan churna®' using ICH guidelines. The use of Accelerated Solvent Extraction (ASE) as an alternative to conventional techniques has been explored. The methanol extracts of root, its formulation and both standard solutions were applied on silica gel F254 HPTLC plates. The plates were developed in Twin chamber using mobile phase toluene: ethyl acetate (6:4, v/v) and scanned at 342 and 325 nm (λmax of piperine and piperlongumine, respectively) using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of piperine and piperlongumine in the range of 20-100 and 30-150 ng/spot, respectively; the correlation coefficient was 0.9957 and 0.9941 respectively. Sharp, symmetrical and well resolved peaks of piperine and piperlongumine spots resolved at Rf 0.51 and 0.74, respectively from other components of the sample extracts. The HPTLC method showed good linearity, recovery and high precision of both markers. Extraction of plant using ASE and rapid HPTLC method provides a new and powerful approach to estimate piperine and piperlongumine as phytomarkers in the extract as well as its commercial formulations for routine quality control.

Quantitative estimation of piperine in piper nigrum and piper longum using high performance thin layer chromatography.

Piperine, a characterizing compound present in fruits of Piper nigrum and Piper longum used as bioavailability enhancer. Ingredients of antioxidant, anti-inflammatory activity has been extracted using soxhlet and supercritical fluid extraction technique. It was isolated using column chromatography. Characterization of compound was done by spectroscopic technique. A simple, rapid, accurate and specific HPTLC method developed and validated. The method proposed can be used for the routine analysis of both P.nigrum and P.longum fruit material and its formulations.

Antimicrobial efficacy of fruit extracts of two piper species against selected bacterial and oral fungal pathogens

Aim: To assess the antimicrobial efficacy of five solvent extracts of two Piper species commonly used in diet and traditional medicine, P. cubeba and P. longum, against selected bacterial and oral fungal pathogens i.e. Streptococcus mutans, Staphylococcus aureus, Candida albicans and Saccharomyces cerevisiae. Methods: The antimicrobial activity of five extracts of cubeb berries and Indian long pepper fruits was determined by the agar well diffusion method. The minimum inhibitory concentration (MIC) for the acetonic, methanolic and ethanolic extracts was determined by the modified agar well diffusion method. Results: Of the 5 fruit extracts evaluated, acetone, ethanol and methanol extracts of both the Piper spp. were found to have variable antimicrobial activities against all the four oral pathogens. The acetonic fruit extract of P. cubeba was the most effective against both the yeasts with the highest zone of inhibition (15.31 mm) against C. albicans followed by the methanolic (12.31 mm) and ethanolic (11.94 mm) extracts. C. albicans was found to be most sensitive pathogen, which survived up to 6.25 mg/mL in the acetonic extract (MIC = 12.5 mg/mL) followed by the methanolic and ethanolic extracts (MIC = 25 mg/mL). The acetonic, methanolic and ethanolic extracts of P. longum fruits showed almost equal inhibition zones of both yeasts, ranging between 10.64 and 14 mm. C. albicans survived up to 12.5 mg/mL (MIC= 25 mg/mL) while S.cerevisiae survived up to 25 mg/mL (MIC = 50 mg/mL). Conclusions: The crude extracts obtained from the fruits of the two Piper spp. may be used to treat oral fungal species, especially C. albicans, as they produced larger inhibition zones than antifungal drugs often used to treat these pathogens.

Bioenhancers: Revolutionary concept to market.

Treatment cost is a major concern for modern medicine in developing countries like India and systematic innovative means to reduce these costs are needed. This article reviews the concept of bioenhancers to reduce treatment costs by increasing drug bioavailability. This concept, based on the Ayurvedic system of medicine, works for a wide range of ingested substances, and has been applied to modern drugs, particularly single chemicals. It offers a fine example of the benefit of integrating an ancient system with modern medicine in both theory and practice.

Free Radical Scavenging Action of Volatile Oil from Piper longum L.and Its Relationship with Molecular Structure / 中药新药与临床药理

Objective To observe the free radical scavenging action of volatile oil from Piper longum L.growing in Hainan Province and to explore the relationship of the action with molecular structure.Methods 1,1-Diphenyl-2-picrylhydrazyl radical(DPPH) method was used to determine the scavenging action.The components of volatile oil were analyzed by GC-MS,and their molecular structures were identified by databank.Results(1)The volatile oil from Piper longum L.had scavenging actions on free radicals in a concentration-dependent manner.(2)Forty one compounds were found in volatile oil from Piper longum L.growing in Hainan Province,in which the amount of carbon-carbon double bond(C=C) was in predomination.Conclusion Carbon-carbon double bond(C=C)is the essential functional group for free radical scavenging action of volatile oil from Piper longum L.

EFFECTS OF OIL OF PIPER LONGUM UNSAPONIFIABLE MATTER ON CHOLESTEROL METABOLISM IN EXPERIMENTAL HYPERCHO-LESTEROLEMIC MICE / 中国药理学通报

Exogenous hypercholesterolemia in mice was induced by feeding with diets containing 2% cholesterol. It was found that Oil of Piper Longum Unsaponifiable Matter ( OPUM ) at 40mg/kg?d-1 ig for 20d could reduce the levels of total cholesterol (TC). ( LDL + VLDL) - c and the hepatic cholesterol content, increase that of biliary cholesterol in exogenous hypercholesterolemic mice. The hypocholesterolemic effect of OPUM is in a dose-dependent manner over the range of 20~40mg/kg. OPUM at 40mg/kg ig could elevate Lecithin - Cholesterol Acyltransferase ( LCAT ) activity inhibited by treating with ip of yolk. It was suggested that the hypochole-sterol effect of OPUM could be concerned with protectiong cholesterol esterification and be of great advantage to protection from arteriosclerosis arteriosclerosis in mice.