Research progress in chemical constituent and pharmacological activity of Punica granatum L / 中国药科大学学报

@#Punica granatum L.(pomegranate) is a medicinal plant belonging to the genus Punica Linn..The peel, seed, flower, leaf and root of P.granatum is widely used as traditional medicine in China.Phytochemical studies showed that the major chemical constituents of P.granatum were tannins, flavonoids, terpenes, alkaloids, phenolic acids, anthocyanins, fatty acids, etc.Biological studies on extracts and active ingredients of P.granatum show some bioactivities, such as antioxidant, hypoglycemic, anti-inflammatory, anti-tumor, antibacterial activities.Herein, the chemical constituents and pharmacological effects of different parts of pomegranate were reviewed, providing a theoretical basis for the further research and utilization of pomegranate.

Extratos hidroetanólicos de canela (Cinnamomum verum) e romã (Punica granatum L) promovem ação antibacteriana frente cepas resistentes a antibióticos de Acinetobacter baumannii e Pseudomonas aeruginosa / Cinnamon (Cinnamomum verum) and pomegranate (Punica granatum L) hydroethanolic extracts promote antibacterial action against antibiotic-resistant strains of Acinetobacter baumannii and Pseudomonas aeruginosa

A resistência bacteriana tem aumentado progressivamente no mundo, assim, há necessidade de novas opções de tratamentos. A fitoterapia tem ganhado notoriedade para combater infecções, principalmente as causadas por bactérias resistentes aos antibacterianos disponíveis. Diante do exposto, o presente estudo teve como objetivo preparar e analisar a composição fitoquímica e a ação antibacteriana dos extratos hidroetanólicos de canela (EHC) e romã (EHR) isolados e associados frente culturas planctônicas e biofilmes de cepas padrão e clínicas de Acinetobacter baumannii e Pseudomonas aeruginosa, além disso, analisar a ação citotóxica dos extratos em queratinócitos humanos (HaCat). Para isso, os EHC e EHR foram preparados e quantificado o teor de sólidos solúveis. Posteriormente, foi quantificado o teor de flavonoides e fenóis totais, análise antioxidante por meio da redução do radical 2,2'-difenil-1-picrilhidrazila (DPPH), e a fitoquímica por cromatografia líquida (HPLC). Em relação a ação antibacteriana dos extratos, foi aplicado o teste de microdiluição em caldo (CLSI ­ M7-A9) e a ação sinérgica realizada por meio do ensaio de checkerboard. As concentrações mais efetivas foram analisadas sobre biofilmes em formação (prevenção) e biofilmes formados (tratamento de 24 h), e quantificada a viabilidade por meio do teste colorimétrico MTT. Para avaliar a citotoxidade, os tratamentos foram aplicados sobre cultura celular de HaCat por 24 h e analisados por meio do teste colorimétrico MTT. A análise estatística foi realizada com 5% de significância (p<0.05), analisados pelo método ANOVA complementado pelo Teste de Tukey. Os resultados demonstraram que os EHC e EHR possuem ação antioxidante e presença de fitocompostos. Os extratos apresentaram ação antibacteriana para todas as cepas avaliadas, quando os mesmos foram associados, obteve-se concentrações sinérgicas para as cepas clínicas de A. baumannii. Em relação a ação antibiofilme, o EHC inibiu a formação em 95% e EHR em 96% do biofilme de #Ab 1, enquanto a cepa #Pa 2 teve 92% e 93% de inibição quando em contato com EHC e EHR, respectivamente. Após tratamento de 24 h em biofilmes formados, as reduções da viabilidade foram de 72% para as cepas #Ab 2 e #Ab 3 quando em contato com o EHC, já EHR inibiu em 83% a viabilidade da cepa #Ab ATCC. Para P. aeruginosa (#Pa 2), as reduções da viabilidade foram de 84% e 88,5% quando tratados com EHC e EHR, respectivamente. A avaliação da citotoxicidade em HaCat demonstrou que após tratamentos com diferentes concentrações dos extratos a viabilidade celular se manteve acima de 70% em todos os grupos. Diante disso, conclui-se que os EHC e EHR apresentam importante ação antioxidante e antibacteriana, tanto em culturas planctônicas quanto em biofilmes, e não apresentaram efeitos citotóxicos na faixa de concentração testada. (AU)

Bacterial resistance has progressively increased in the world, thus, there is a need for new treatment options. Phytotherapy has gained notoriety for fighting infections, mainly those caused by bacteria resistant to available antibacterials. In view of the above, the present study aimed to prepare and analyze the phytochemical composition and antimicrobial action of hydroethanolic extracts of cinnamon (EHC) and pomegranate (EHR) isolated and associated against planktonic cultures and biofilms of standard and clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition, analyze the cytotoxic action of the extracts on human keratinocytes (HaCat). For this, the EHC and EHR were prepared and the soluble solids content was quantified. Subsequently, the content of flavonoids and total phenols, antioxidant analysis through the reduction of the radical 2,2'-diphenyl1-picrylhydrazyl (DPPH), and phytochemistry by liquid chromatography (HPLC) were quantified. Regarding the antimicrobial action of the extracts, the broth microdilution test (CLSI ­ M7-A9) was applied and the synergistic action was performed through the checkerboard test. The most effective concentrations were analyzed on forming biofilms (prevention) and formed biofilms (24 h treatment), and viability was quantified using the MTT colorimetric test. To evaluate the cytotoxicity, the treatments were applied on HaCat cell culture for 24 h and analyzed using the MTT colorimetric test. Statistical analysis was performed with 5% significance (p<0.05), analyzed by the ANOVA method complemented by the Tukey test. The results showed that the EHC and EHR have antioxidant action and presence of phytocompounds. The extracts showed antibacterial action for all evaluated strains, when they were associated, synergistic concentrations were obtained for the clinical strains of A. baumannii. Regarding the antibiofilm action, EHC inhibited formation by 95% and EHR by 96% of the #Ab 1 biofilm, while the #Pa 2 strain had 92% and 93% inhibition when in contact with EHC and EHR, respectively. After 24 h treatment in formed biofilms, viability reductions were 72% for strains #Ab 2 and #Ab 3 when in contact with EHC, whereas EHR inhibited the viability of strain #Ab ATCC by 83%. For P. aeruginosa (#Pa 2), viability reductions were 84% and 88.5% when treated with EHC and EHR, respectively. The evaluation of cytotoxicity in HaCat showed that after treatments with different concentrations of extracts, cell viability remained above 70% in all groups. Therefore, it is concluded that EHC and EHR have important antioxidant and antibacterial action, both in planktonic cultures and in biofilms, and did not show cytotoxic effects in the tested concentration range. (AU)

Anti-Obesity Effect of Methanolic Extracts of Local Punica granatum in High-Fat Diet-Induced Obese Rats

@#Introduction: Obesity has been linked to coronary artery disease, type II diabetes, metabolic syndrome, stroke, and cancer. Pomegranate (Punica granatum; PG) has been used extensively in folk medicine for several therapeutic purposes. The goal of this study is to investigate the anti-obesity effect of PG peel and pulp methanolic extracts in highfat diet (HFD)-induced Sprague-Dawley rats. Methods: The animals were separated into 7 groups namely Normal control group (normal diet); HFD-induced only as negative control group, and positive control group (HFD-induced + orlistat); treatment group included HFD-induced + peel 125 mg/kg, HFD-induced +peel 250 mg/kg, HFD-induced + pulp 125 mg/kg and HFD-induced + pulp 250 mg/kg. Results: It was observed that methanolic extract of peel and pulp PG 250 mg/kg showed low increment of body weight with a reduction in weight of liver, visceral fat, and subcutaneous fat. Compared to the negative control group, total cholesterol, triglyceride, and low-density lipoprotein levels were shown to be lowered for PG peel and pulp groups also showed higher values in high-density lipoprotein. Conclusion: PG reduced obesity-related markers in blood, liver, and adipose tissue and inhibited obesity caused by a high-fat diet probably because of its antioxidant properties.

An aqueous pomegranate peel extract (Punica granatum) protect against Elastase-induced pulmonary emphysema in Sprague Dawley rats model

We investigated the effect of Punica granatum peel aqueous extract (PGE), on pulmonary inflammation and alveolar degradation induced by intratracheal administration of Elastase in Sprague Dawley rats. Lung inflammation was induced in rats by intratracheal instillation of Elastase. On day 1 and 2, animals received an intraperitoneal injection of PGE (200 mg/mL), three hours later, they were intratracheally instilled with 25U/kg pancreatic porcine Elastase. Animals were sacrificed 7 days later. Bronchoalveolar lavage (BAL) were collected and cellularity, histology and mRNA expression of Monocyte chemotactic protein 1(MCP-1), Tumor Necrosis Factor-Alpha (TNF-α), Interleukin 6 (IL-6), and Matrix Metalloproteinase-2 (MMP-2) were studied. In addition, activity of TNF- α, IL-6 and MCP-1 on BAL were also analyzed by ELISA Kit. Elastase administration increased: BAL cellularity, neutrophils recruitment and BAL MCP1, IL-6 expressions. It also increased lung TNF-α, MCP-1, MMP-2 expressions, platelets recruitment, histological parameters at 7th day of elastase treatment. Intraperitoneal injection of 200 mg/kg of PGE reduced, significantly, BAL cellularity, and neutrophils recruitment. However, in animal treated with PGE, MCP-1, MMP-2 and IL-6 on day 7, were similar to the Sham group. Treatment with PGE (200 mg/ kg) also significantly reduced lung TNF-α, and MCP-1 expression. This study reveals that PGE Punica granatum protects against elastase lung inflammation and alveolar degradation induced in rats

Genetic diversity among wild pomegranate (Punica granatum) in Azad Jammu and Kashmir region of Pakistan

BACKGROUND: Pomegranate (Punica granatum L.), one of the most important tropical fruits in Azad Jammu and Kashmir regions of Pakistan, is highly valued for its nutrition and medicinal purposes. Although pomegranate is native to this region, the genetic diversity among wild pomegranate accessions is currently unknown. Such information would be vital for germplasm conservation and breeding efforts. In the current study, genetic diversity among forty-eight wild pomegranate accessions collected from different agro-ecological zones of Azad Jammu and Kashmir was assessed using 41 simple sequence repeat (SSR) markers. RESULTS: The markers revealed 303 alleles averaging 7.39 alleles per marker. Polymorphic information content ranged from 0.12 (PGCT093B) to 0.88 (Pom006), with a mean of 0.54. The average genetic distance (GD) across all genotypes was 0.52, and was lowest between Chattar Class and Thorar genotypes (GD = 0.27), but highest between Khun Bandway and Akhor Ban (GD = 0.74). A neighbor-joining dendrogram separated the genotypes into three major clusters, with further sub-clustering within each cluster. CONCLUSIONS: Overall, the results presented here show significant genetic diversity among wild pomegranate accessions in Azad Jammu and Kashmir region of Pakistan. These accessions present a valuable genetic resource to breeding and cultivar improvement programs within the region.

Evaluations of antidepressant activity of Punica granatum peel extract in albino mice

Background: Depression is the most common disorder of mental illnesses and affects excess of 10-15% of population. According to the WHO reports, more than 350 million persons suffer from depression all over the world. The aim of present study is to evaluate anti-depressant activity of Punica granatum peel extract (PgPE) in albino mice.Methods: Male albino mice (20-30 g) were used. Animals were divided into 5 groups with 6 animals in each which were subjected to forced swim test. Group 1 is control, group 2 received (standard) imipramine 10 mg/kg, p.o, group 3 (T1) PgPE 50 mg/kg, p.o, group 4 (T2) PgPE 100 mg/kg, p.o, group 5 (T3) PgPE 200 mg/kg, at first animals were forced to swim for 15 min (trained), and the study was performed after 24 hrs. All the animals were treated with individual drug 60 min prior to study, animals were forced to swim for 6 min and the duration of immobility was recorded. The mouse was considered immobile when it floats motionlessly or made only those moments necessary to keep its head above the water surface. The total duration of immobility of each mouse was recorded after the test in each group.Results: The duration of immobility is significantly reduced at PgPE 200 mg/kg and results were analysed by one way analysis of variance (ANOVA).Conclusions: PgPE 200 mg/kg significantly (p<0.05) decreases the duration of immobility in mice.

Punica granatum improves renal function in gentamicin-induced nephropathy in rats via attenuation of oxidative stress

Background: Gentamicin is widely used as an antibiotic for the treatment of gram negative infections.Evidences indicates that oxidative stress is involved in gentamicin-induced nephrotoxicity. In Ayurvedicmedicine, Punica granatum Linn. is considered as 'a pharmacy unto itself”. It has been claimed intraditional literature, to treat various kidney ailments due to its antioxidant potential.Objective: To explore the possible mechanism of action of methanolic extract of P. granatum leaves(MPGL) in exerting a protective effect on gentamicin-induced nephropathy.Material and methods: Animals were administered with gentamicin (80 mg/kg/day i.m.) and simultaneously with MPGL (100, 200 and 400 mg/kg p.o.) or metformin (100 mg/kg p.o.) for 8 days. A satellitegroup was employed in order to check for reversibility of nephrotoxic effects post discontinuation ofgentamicin administration. At the end of the study, all the rats were sacrificed and serumeurine parameters were investigated. Antioxidant enzymes and tumor necrosis factor alpha (TNF-a) levels weredetermined in the kidney tissues along with histopathological examination of kidneys.Results: Increase in serum creatinine, urea, TNF-a, lipid peroxidation along with fall in the antioxidantenzymes activity and degeneration of tubules, arterioles as revealed by histopathological examinationconfirmed the manifestation of nephrotoxicity caused due to gentamicin. Simultaneous administration ofMPGL and gentamicin protected kidneys against nephrotoxic effects of gentamicin as evidenced fromnormalization of renal function parameters and amelioration of histopathological changes.Conclusion: Data suggests that MPGL attenuated oxidative stress associated renal injury by preservingantioxidant enzymes, reducing lipid peroxidation and inhibiting inflammatory mediators such as TNF-a.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Os efeitos da Punica granatum L. em diferentes concentrações sobre duas linhagens celulares: estudo in vitro / The effects of Punica granatum L. at different concentrations on two cell lines: in vitro study

Introdução: A Punica granatum L. (PG), utilizada como medicamento fitoterápico, apresenta propriedades terapêuticas, anti-inflamatórias e antioxidante. Embora diversos estudos já tenham sido realizados com este fitoterápico, seus possíveis efeitos citotóxicos nos tecidos humanos ainda não são claros. Objetivo: Avaliar a citotoxicidade da PG por meio de cultura celular com duas linhagens: fibroblastos humanos de mucosa oral (FLM) e células de carcinoma epidermoide oral humano (KB). Material e método: As células foram submetidas ao teste de viabilidade celular por 24 horas nas concentrações da PG 1%, 0,50%, 0,25%, 0,125%, 0,062% e 0,031%, e aos testes de citotoxicidade celular em 4 horas, 1, 3, 5 e 7 dias, em diferentes concentrações, realizados em triplicata. Foi utilizado um controle negativo (Triton 1%) e um controle positivo sem o extrato de PG. Os dados obtidos foram submetidos à ANOVA (p < 0,05). Resultado: Foi possível observar que o extrato da PG possui efeitos inibitórios, apresentando-se maior nas células KB em relação às FLM. Os testes de citotoxicidade e viabilidade mostraram inibição e morte das células KB e FLM nas concentrações 1%, 0,50% e 0,25%. Conclusão: O efeito inibitório da PG foi dose-dependentes, mostrando-se maior nas células KB em relação às FLM.

Introduction: Punica granatum L. (PG), used as a herbal medicine, has therapeutic, anti-inflammatory and antioxidant properties, although several studies have already been carried out with this herbal medicine, its possible cytotoxic effects on human tissues are still unclear. Objective: To evaluate the cytotoxicity of PG through cell culture with two strains: human oral mucosa fibroblasts (LFM) and human oral squamous cell carcinoma (KB) cells. Material and method: The cells were submitted to the cell viability test for 24 hours in the concentrations of PG 1%, 0.50%, 0.25%, 0.125%, 0.062% and 0.031% and the cell cytotoxicity tests in 4 hours, 1, 3, 5 and 7 days in different concentrations, performed in triplicate. A negative control (Triton 1%) and a positive control without the PG extract were used. The data obtained were submitted to ANOVA (p <0.05). Result: it was possible to observe that the PG extract has inhibitory effects, being higher in KB cells in relation to LFM. The cytotoxicity and viability tests showed inhibition and death of KB and FLM cells at concentrations of 1%, 0.50% and 0.25%. Conclusion: The inhibitory effect of PG was dose dependent, showing itself to be greater in KB cells compared to LMB.

Extrato da casca de Punica granatum potencializa o efeito anti cárie de um enxaguatório bucal contendo trimetafosfato de sódio e flúor / Punica granatum peel extract enhances the anti caries effect of mouthwash containing sodium trimetaphosphate and fluoride

A cárie dentária está entre as principais e mais comuns doenças bucais. É causada por ácidos produzidos pelo biofilme microbiano que levam à desmineralização do esmalte. A prevenção e controle dessa doença crônica consistem na desorganização periódica do biofilme e na promoção da remineralização dentária. Para resolver esse problema, associamos o extrato de casca de Punica granatum (romã) (PPE) ao trimetafosfato de sódio (TMP) e fluoreto (F) em formulações para uso como enxaguatório bucal, e avaliamos sua eficácia na redução do processo de desmineralização do esmalte dental, bem como seu potencial anti biofilme contra importantes patógenos orais presentes na cárie dentária (Streptococcus mutans ATCC 25175 e Candida albicans ATCC 10231). Blocos de esmalte bovino (4 mm × 4 mm) selecionados por dureza superficial inicial (SHi) foram alocados aleatoriamente de acordo com grupos de tratamentos de formulação (n = 12 / grupo): ETF1 (3,0% PPE + 0,2% TMP + 100ppmF), TF1 (0,2% TMP + 100ppmF), ETF2 (3,0% PPE + 0,3% TMP + 225ppmF), TF2 (0,3% TMP + 225ppmF), F1 (100 ppmF), F2 (225 ppmF) e P (formulação sem E / T / F - placebo). Os blocos foram tratados 2x / dia com cada formulação e submetidos a cinco ciclos de pH (soluções desmineralizantes / remineralizantes) a 37° C. A seguir, determinaram-se a dureza superficial final (SHf), a dureza integrada da subsuperfície de perda (ΔKHN) padronizada e as concentrações de fluoreto de esmalte (F) de cálcio (Ca) e fósforo (P). A porcentagem de perda de dureza superficial (% SH) foi calculada (% SH = [(SHf - SHi) / SHi)] x 100), e as formulações que promoveram menores porcentagens de desmineralização do esmalte (% SH) e suas contrapartes foram selecionadas para os ensaios anti biofilme, bem como a formulação contendo apenas PPE (E). Para isso, ETF2 e TF2 (% SH = -34,5% e -53,1%, respectivamente), e a formulação E foram usadas para tratar por 1 ou 10 minutos biofilmes duplos de C. albicans e S. mutans crescidos por 24 horas em discos de hidroxiapatita (HA). A desmineralização da superfície do esmalte foi menor nas amostras tratadas com a formulação ETF2, resultando em uma diminuição de 46% na% SH em comparação com a F2. Novamente, a capacidade de reduzir o corpo da lesão (ΔKHN) foi maior (~ 26%) com ETF2 em relação a F2, e F2 proporcionou a maior concentração de F, Ca e P na superfície do esmalte. Entre as formulações de enxaguatório bucal ETF2, TF2 e E, as maiores taxas de redução de células viáveis foram exibidas tratando o biofilme com ETF2 por 10 minutos, independentemente do microrganismo testado. Em conclusão, a adição de PPE (3%) em enxaguatórios bucais contendo TMP (0,3) e F (225ppm) promoveu uma diminuição considerável no mineral sem perda de esmalte dental, além de reduzir consideravelmente o biofilme cariogênico formado por S. mutans e C albicans. Assim, cria uma perspectiva promissora para o desenvolvimento de um produto comercial dental sem álcool com os benefícios de milênios reconhecidos à saúde do Punica granatum(AU)

Dental caries is among the main and most common oral diseases. It is caused by acids produced by microbial biofilm that lead to enamel demineralization. The prevention and control of this chronic disease consist of periodic disorganization of the biofilm and the promotion of dental remineralization. To address this problem, we associate Punica granatum (pomegranate) peel extract (PPE) with sodium trimemtaphosphate (TMP), and fluoride (F) in formulations for being used as mouthwash, and evaluate its efficacy on reducing dental enamel demineralization process as well as its antibiofilm potential against important oral pathogens present in dental caries (Streptococcus mutans ATCC 25175 and Candida albicans ATCC 10231). Bovine enamel blocks (4 mm × 4 mm) selected by initial surface hardness (SHi) were randomly allocated according to groups of formulation treatments (n= 12/group): ETF1 (3.0%PPE+0.2%TMP+100ppmF), TF1 (0.2%TMP+100ppmF), ETF2 (3.0%PPE+0.3%TMP+225ppmF), TF2 (0.3%TMP+225ppmF), F1 (100 ppmF), F2 (225 ppmF), and P (formulation without E/T/F - placebo). The blocks were treated 2×/day with each formulation and submitted to five pH cycles (demineralizing/remineralizing solutions) at 37°C. Next, final surface hardness (SHf), integrated loss subsurface hardness (ΔKHN), and enamel fluoride (F) calcium (Ca) and phosphorus (P) concentrations were determined. The percentage of surface hardness loss (%SH) was calculated (%SH = [(SHf - SHi)/SHi)] x 100), and the formulations which promoted lower percentages of enamel demineralization (%SH) and its counterparts were selected to the antibiofim assays, as well as formulation containing only PPE (E). For that, ETF2 and TF2 (%SH= -34.5% and -53.1% respectively), and formulation E were used to treat for 1 or 10 minutes dual biofilms of C. albicans and S. mutans grown for 24 hours on hydroxyapatite discs (HA). Demineralization of the enamel surface was lower in samples treated with formulation ETF2, resulting in a 46% decrease in %SH in comparison with F2. Again, the capacity to reduce the lesion body (ΔKHN) was higher (~ 26%) with ETF2 in relation to F2, and F2 provided the highest concentration of F, Ca and P in enamel surface. Amongst the mouthwash formulations ETF2, TF2 and E, the highest rates of viable cells reduction were exhibited by treating biofilm with ETF2 for 10 minutes regardless of the microorganism tested. In conclusion, the addition of PPE (3%) in mouthwashes containing TMP (0.3) and F (225ppm) promoted a considerably decrease in the mineral loss of dental enamel besides considerable reducing cariogenic biofilm formed by S. mutans and C. albicans. It thus creates a promising prospect for the development of an alcohol free dental commercial product with the millennial recognized health benefits of Punica granatum(AU)

Study on Medication Regulation and Pharmaceutical Property of Tibetan Medicine Prescription of Punica gran- atum Seeds Based on Apriori Algorithm and “Ro Nus Zhu rJes ”Vector Model / 中国药房

OBJECTIVE：To invest igate the medication regulation of Tibetan medicine prescription of Punica granatum seeds， and to analyze the pharmaceutical property of it in the treatment of “Sanweihuo”（Me drod gusm) dysfunction induced-related diseases. METHODS ：Using Clinical Notes of Tibetan Medicine ，Medicine Standard of the Ministry of Public Health ：Tibetan Medicine as standard ，Tibetan medicine prescriptions of P. granatum seeds recorded in Sibu Yidian ，Yuewang Yaozhen and other medical records were collected ，and TCMISS V 2.5 software was used to analyze main indication and medicine frequency statistically. The core medicine combinations of the prescriptions were analyzed with Apriori algorithm （support degree ≥50％， confidence degree ≥90％)，and medication regulations were analyzed. Tibetan medicine “Ro Nus Zhu rJes ”vector model was used to analyze the properties of Tibetan medicine prescription of P. granatum seeds in the treatment of high frequency diseases ，and K-means cluster analysis was carried out for seventeen effectiveness. RESULTS ：A total of 300 Tibetan medicine prescriptions of P. granatum seeds were collected ，involving 104 kinds of main indications ，among which frequency of 18 kinds of indications was more than or equal to 10 times，including“non-indigestion syndrome ”（Ma Zhu ba) （76 times)，“wood cloth disease ”（Smug po) （46 times)，“edema”（Chu nad) （34 times)，etc. Totally 322 kinds of Tibetan medicines were involved. Among them ，21 flavors were more than or equal to 50 in frequency ，including Piper longum （192 times)，Amomum kravanh （164 times) and Carthamus tinctorius（134 times)，etc. Fifty-six core combinations were obtained ，mainly including P. granatum seeds-P. longum （192 times)， P. granatum seeds-A. kravanh （164 times) and P. granatum seeds-C. tinctorius （134 times). The prescriptions （13 pieces) with six flavors as pungent ，sweet and sour ，and with prominent medicinal properties such as roughness ，heat and dry effects could be used to treat “non-indigestion syndrome ”，and then included into type Ⅰ. The prescriptions （13 pieces) with six flavors as bitter ，sweet and pungent ，and with prominent medicinal properties such as cooling ，dull and heavy effects could be used to treat “wood cloth disease”，and then included into type Ⅱ. The prescriptions （11 pieces) with six flavors as sweet ，sour and pungent ，and with prominent medicinal properties such as dull ，heavy and moistening effects could be used to treat “edema”，and then included into type Ⅲ. CONCLUSIONS ：Tibetan medicine P. granatum seeds is often used in combination with the isothermal medicines such as P. longum ，A. kravanh . P. granatum seeds are also commonly found in cool prescriptions ，revealing that Tibetan medicine pays attention to “Sanweihuo”while treating diseases ，which embodies the academic thought of Tibetan medicine that “there is cooling in temperature ，and there is temperature in cooling ”.

Molecular characterization of PgUFGT gene and R2R3-PgMYB transcription factor involved in flavonoid biosynthesis in four tissues of wild pomegranate (Punica granatum L.)

The diversity on fruit colouration in plants directly depends on the flavonoids that explain the development of different pigmentation patterns. Anthocyanins are the major class of flavonoid pigments that are synthesized through flavonoid biosynthetic pathway. In the present study, two genes: PgUFGT gene and R2R3-PgMYB gene, involved in anthocyanin biosynthesis were analysed in four tissues of wild pomegranate. The structural genes, UDP-glucose: flavonoid-3-O-glucosyl transferase (PgUFGT; GenBank accession number: MK058491) and its myeloblastosis transcription factor (R2R3-PgMYB; GenBank accession number: MK092063) were isolated and their expression pattern were studied. Molecular modelling indicated that the main secondary structures of PgUFGT and R2R3-PgMYB genes are α-helix and random coil. In addition, expression profiling of PgUFGTand R2R3-PgMYB by quantitative-real time PCR indicated a positive correlation between anthocyanin content and their expression in leaves, flowers, green and red fruits of wild pomegranate. Among all the tissues, the red fruit exhibited high transcripts levels of PgUFGT as well as R2R3-PgMYB transcription factor. An extensive homology with UFGTs from other plants was revealed on comparative and bioinformatic analyses. Present study reveals that PgUFGT plays a predominant role in anthocyanin content in wild pomegranate fruits. Further, it is strongly suggested that R2R3-PgMYB transcription factor regulates the anthocyanin biosynthesis in wild pomegranate via expression of PgUFGT gene. This is the first study which provides an insight on expression profile of PgUFGT and R2R3-PgMYB that are involved in colour development and fruit ripening in wild pomegranate.

Evaluation of micro-irrigation methods in pomegranate under semi-arid tropical climate of India

Aim: To evaluate different micro-irrigation systems against basin irrigation in terms of growth, yield, fruit quality, disease infestation and water use of pomegranate under semi-arid tropical climate of Central India. Methodology: A field experiment was conducted in newly planted pomegranate cv. Bhagwa orchard with four irrigation treatments, viz., drip (4 lph, 4 plant-1), microjet 180o (2 plant-1), microjet 360o (2 plant-1) and basin (double ring) irrigation replicated five times in a randomised block design (RBD) in heavy textured soil. Results: Basin irrigation is widely practiced method of irrigation which presently facilitate better soil moisture distribution, resulted in better root proliferation and higher leaf nutrient status (K 0.98%, Cu 105.4 ppm and Zn 35.9 ppm) followed by drip (N 2.12% and Mn 40.4 ppm) system of irrigation. Maximum fruit yield was obtained under basin and drip which was 376.7% and 242.8% higher than that obtained under microjet 180o irrigation system, respectively. Water use efficiency was highest in drip (0.526) than basin (0.499) system of irrigation. Disease incidence in the plants was less under drip while it was high under microjet system of irrigation. Interpretation: Experimental findings clearly indicate that the areas which had ample amount of irrigation, basin irrigation was the most efficient system in harnessing maximum yields, while drip irrigation resulted in higher water use efficiency under arid to semiarid tropical climate, micro-sprinkler irrigation system is not beneficial for pomegranate orchards.

Extract of the Punica granatum Flowers Attenuates Progression of Hepatoxicity against Cadmium Chloride-Induced Liver Injury in Rats

Background: In the present study, we investigated the role of cadmium in acute liver injury, as well as the underlying mechanisms. Methods: On the other hand, we investigated whether Punica granatum flower extract (PG) has a healing effect on CD-induced liver damage. Control groups (G1-G2) received water and 200 mg/kg PG, respectively. Toxicity groups (G3-G6) received 2 different doses of Cd (15-30 mg/kg b.w) orally in a single administration. To evaluate liver function, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyl transferase (GGT) were detected in the serum, superoxide dismutase (SOD), glutathione peroxidase (GPx), thiobarbituric acid reactive substances (TBARS) were established in the liver and 8-hydroxy-20-deoxyguanosine (8-OHdG) levels were determined for the antioxidative and anti inflammatory effects of PG. Tissue sections were also evaluated histopathologically. Results: We found that Cd exposure decreased levels of SOD, GPx and, increased expression of caspase-3 and levels of TBARS and 8-OHdG. Biochemical and histopathological analysis revealed the toxic effects of cadmium on the liver for the rats in oral acute toxicity study. Functional parameters were significantly improved in PG-treated groups and the severity of the liver injury and apoptosis were significantly decreased in this group. Conclusion: As a result, PG can be consumed as a protective agent against acute liver injuries.

Actividad cicatrizante in vivo del polvo carbonizado de Punica granatum Linn y Eichhornia crassipes / In vivo healing activity of the charred powder of Punica granatum Linn and Eichhornia crassipes

Introducción: Las especies vegetales constituyen el remedio primero a los problemas de salud que aquejan a las personas. La granada es rica en ácido ascórbico, mejora la barrera epidérmica y reduce la contracción de las heridas. La flor de agua posee actividad antibacteriana. Objetivo: Evaluar el efecto cicatrizante del polvo carbonizado de Punica granatum Linn (granada) y de Eichhornia crassipes (flor de agua), en un modelo experimental en ratas. Métodos: Estudio analítico experimental con el empleo 30 ratas macho distribuidas en 3 grupos (n= 10). Grupo I y II; tratadas con el polvo carbonizado de la granada y flor de agua respectivamente. Grupo III: Tratadas con cloruro de sodio al 0,9 por ciento. Se realizó un modelo de herida por escisión en el dorso. La evolución de la cicatrización fue seguida por la velocidad de contracción de la herida en milímetros. Se realizó histología sobre biopsias de tejido cicatrizado. Para el análisis estadístico se empleó la prueba no paramétrica de Mann- Whitney. El nivel de significación se fijó en p < 0,05. Resultados: Se comprobó el efecto cicatrizante de los polvos carbonizados de la granada y flor de agua; disminuyó el área de las heridas de manera significativa respecto al grupo control. El estudio histológico mostró dermis madura grado III en los grupos I y II. Conclusiones: La aplicación tópica del polvo carbonizado de granada y de flor de agua influyó sobre el cierre de las heridas y en la maduración de la dermis, por lo cual favoreció la cicatrización(AU)

ABSTRACT Introduction: Plant species are the first remedy to the people's health problems. The pomegranate is rich in ascorbic acid, improves the epidermal barrier and reduces the contraction of wounds. The water hyacinth has antibacterial activity. Objective: To evaluate the healing effect of the charred powder of Punica granatum Linn (pomegranate) and Eichhornia crassipes (water hyacinth), in an experimental model in rats. Methods: Experimental analytical study with 30 male rats distributed in 3 groups (n = 10). Group I and II; treated with the charred powder of the pomegranate and water hyacinth respectively. Group III: treated with 0.9 percent sodium chloride. An incision wound model was made on the back. The evolution of healing was followed by the speed of contraction of the wound in millimeters. Histology was performed on biopsies of scar tissue. For the statistical analysis, the nonparametric Mann-Whitney test was used. The level of significance was set at p < 0.05. Results: The healing effect of the charred powders of the pomegranate and water hyacinth was verified; decreased the wound area significantly compared to the control group. The histological study showed mature grade III dermis in groups I and II. Conclusions: The topical application of the charred powder of pomegranate and water hyacinth influenced the closure of the wounds and the maturation of the dermis, which favored healing(AU)

Evaluación de la actividad inmunomoduladora del polvo seco de Punica granatum Linn en un modelo experimental de inmunosupresión / Evaluation of the immunomodulatory activity of dry powder of Punica granatum Linn in an experimental model of immunosuppression

Introducción: La inmunosupresión es uno de los principales obstáculos en el tratamiento de cáncer, por esta razón, diversos inmunomoduladores naturales y sintéticos son estudiados, con el fin de atenuar los efectos de la terapia convencional. Objetivo: Evaluar la actividad inmunomoduladora del polvo seco de Punica granatum Linn (granada). Métodos: Ratas machos Wistar fueron divididas en cuatro grupos: grupo I (control), II (inmunizado con glóbulos rojos de carnero), III (inmunosuprimido con ciclofosfamida e inmunizado con glóbulos rojos de carnero) y IV (tratado con polvo de granada, inmunosuprimido con ciclofosfamida e inmunizado con glóbulos rojos de carnero). Las variables estudiadas fueron el peso corporal y detección de anticuerpos hemoaglutinantes. Resultados: Se observó ligera tendencia al aumento del peso corporal en los grupos I, II y III, con respuesta diferente en el grupo IV, donde hubo una leve disminución. El título de anticuerpos del grupo III disminuyó con respecto al II y IV, tanto en la respuesta primaria como secundaria. En el grupo IV el título de anticuerpos resultó ser estadísticamente significativo con relación al del grupo III (p= 0,000) para ambas respuestas. Conclusiones: La Púnica granatum mostró efecto inmunomodulador, al incrementar el nivel de anticuerpos hemaglutinantes(AU)

Introduction: Immunosuppression is one of the main obstacles in the treatment of cancer, for this reason, several natural and synthetic immunomodulators are studied, in order to attenuate the effects of conventional therapy. Objective: To evaluate the immunomodulator activity of Punica granatum Linn (pomegranade). Methods: Wistar male rats were divided into four groups: group I (control), II (immunized with red blood cells of sheep), III (immunosuppressed with cyclophosphamide and immunized with red blood cells of sheep) and IV (treated with pomegranate powder, immunosuppressed with cyclophosphamide and immunized with red blood cells of sheep). The variables studied were body weight and detection of haemagglutinating antibodies. Results: A slight tendency to increase body weight was observed in groups I, II and III, with a different response in group IV, where there was a slight decrease. The antibody titre of group III decreased with respect to II and IV, both in the primary and secondary response. In group IV the antibody titer was found to be statistically significant in relation to group III (p = 0.000) for both responses. Conclusions: Punic granatum showed immunomodulatory effect, increasing the level of haemagglutinating antibodies(AU)

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

Preventive and Curative Effects of Metformin, Nigella sativa, Punica granatum and Zingiber officinale on Male Reproductive Dysfunction in Diabetic Rats

Diabetes mellitus is a metabolic disorder, that affects almost all the cells of the body. Well documented complications of DMinclude, neuropathy, nephropathy, microangiopathy, and retinopathy. The negative impact of the disorder is also found inother cells, including male reproductive system. In this study, fifty adult Wistar albino male rats were used. The rats weredivided into 10 groups, each group comprised of 5 rats. Diabetes was induced by intraperitoneal injection of STZ, effects ofDM were observed on the sperm producing cells then preventive as well as curative effects of Metformin, Punica granatum,Nigella sativa and Zingiber officinale were observed. It was found that all the substances prevent as well as repair/cure thedamage caused by DM to the seminiferous tubules and other structures. The results of this study show that Metforminthough prevented and caused repair to the damaged cells as well, but was not as effective as Punica granatum, Nigellasativa and Zingiber officinale.

Mature pomegranate recognition methods in natural environments using machine vision / Métodos de reconhecimento de romã madura em ambientes naturais usando visão de máquina

ABSTRACT: The use of machine vision to recognize mature pomegranates in natural environments is of major significance in improving the applicability and work efficiency of picking robots. By analyzing the color characteristics of color images of mature pomegranates under different illumination conditions, the feasibility of the YCbCr color model for pomegranate image recognition under different illumination conditions was proven. First, the Cr component map of pomegranate image is selected and then the pomegranate fruit is segmented by the kernel fuzzy C-means clustering algorithm to obtain the pomegranate image. Contrast experiments of pomegranate image segmentation under different illumination conditions were then performed using the proposed kernel fuzzy C-means clustering algorithm, the fuzzy C-means clustering algorithm, the Otsu algorithm and the threshold segmentation algorithm. Results of the experiments verified the effectiveness and superiority of the proposed algorithm.

RESUMO: O uso de máquina para reconhecer romãs maduras em ambientes naturais é de grande importância para melhorar a aplicabilidade e a eficiência do trabalho de robôs de colheita. Ao analisar as características de cor das imagens coloridas de romãs maduras sob diferentes condições de iluminação, a viabilidade do modelo de cores YCbCr para o reconhecimento de imagens de romãs sob diferentes condições de iluminação foi comprovada. Primeiro, o mapa do componente Cr da imagem da romã é selecionado e, em seguida, o fruto da romãzeira é segmentado pelo algoritmo de agrupamento C-means fuzzy do kernel para obter a imagem da romã. Experimentos contrastados de segmentação de imagens de romã sob diferentes condições de iluminação foram então realizados usando o algoritmo proposto de agrupamento C-means fuzzy, o algoritmo fuzzy de agrupamento C-means, o algoritmo Otsu e o algoritmo de segmentação de limiares. Os resultados dos experimentos verificaram a efetividade e superioridade do algoritmo proposto.

Antiurobacterial Activity of Punica granatum L. Seed Extract

The natural products derived from medicinal plants are proven to be the abundant source of biologically active compounds, which have been the basis of development of new pharmaceuticals. The present study was carried out to understand the antibacterial activity of methanolic extract of Punica granatum L. seed (MPG) against the clinically isolated urinary tract infections (UTI) causing bacteria i.e. Escherichia coli and Klebsiella pneumoniae using different methods i.e. disc diffusion, agar well diffusion, modified agar well diffusion. Colony forming units per ml (CFU/ml) were also determined for wild and MPG treated bacteria by spread plate and absorbance method. Phytochemical analysis of MPG and biochemical characterization of K. pneumoniae and E. coli was carried out using a standard protocol. MPG was a dark red colour sticky mass with a yield of 26.87% (w/w). Phytochemical analysis revealed the presence of steroids, tannins, coumarins, glycosides, flavonoids and phenolic compounds. The present investigation showed that MPG was effective against K. pneumoniae and E. coli. K. pneumoniae was a multi-drug resistant strain, found to be more sensitive to MPG in comparison to E. coli. The CFU/ml was found to be reduced in a dose-dependent manner in MPG treated urobacteria, which suggests that P. granatum seeds can be used as a remedy for UTI and also can be taken regularly to prevent the recurrence of UTI.

Studies on Green Synthesis of Copper Nanoparticles Using Punica granatum

Aim: The present study aimed at green synthesis of copper nanoparticles using various plant extracts as reducing and stabilizing agents. It would also study the antibacterial effect of the synthesized copper nanoparticles. Place and Duration of Study: Department of Microbiology, Bhavan’s Vivekananda college of Science, Humanities and Commerce, Hyderabad, India. The duration of the study is for six months between May 2017 to October 2017. Methodology: The aqueous solutions of different plant extracts were mixed with CuSO4 solution and incubated for green synthesis of stable copper nanoparticles. These were tested by UV-Visible spectroscopy and SEM analysis. Antibacterial tests of the biosynthesized nanoparticles were carried out on Gram-positive Bacteria Staphylococcus aureus by Agar well assay. Results: The aqueous solutions of different plant extracts yielded stable copper nanoparticles as indicated by the O.D values tested using UV-Visible spectroscopy. The best plant extract that yielded higher amount of copper nanoparticles was fruit rind extract of Punica granatum. The synthesized nanoparticles were found to be 56-59 nm, characterized by Scanning Electron Microscopy (SEM). The synthesized copper nanoparticles exhibited a strong antibacterial activity against Staphylococcus aureus. Conclusion: The copper nanoparticles can be green synthesized using fruit rind extract of Punica granatum and these can be used as efficient antimicrobial agents against Staphylococcus aureus and the study is significant currently as drug resistant infections of Staphylococcus aureus are gaining much prevalence and prominence.