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In vivo antimalarial activity and toxicological effects of methanolic extract of Cocos nucifera (Dwarf red variety) husk fibre.

Elizabeth Abidemi BALOGUN ; Sylvia Orume MALOMO ; Joseph Oluwatope ADEBAYO ; E-mail: TOPEBAYO2002@YAHOO.COM. ; Ahmed Adebayo ISHOLA ; Ayodele Olufemi SOLADOYE ; Lawrence Aderemi OLATUNJI ; Olatunji Matthew KOLAWOLE ; Stephen Olubunmi OGUNTOYE ; Abiola Samuel BABATUNDE ; Oluwole Busayo AKINOLA

Journal of Integrative Medicine.2014;12(6):504-511. doi:10.1016/S2095-4964(14)60054-6

OBJECTIVEPhytochemical constituents as well as antimalarial and toxicity potentials of the methanolic extract of the husk fibre of Dwarf Red variety of Cocos nucifera were evaluated in this study.

METHODSThe dried powdered husk fibre was exhaustively extracted with hexane, ethyl acetate and methanol successively and the methanolic extract was screened for flavonoids, phenolics, tannins, alkaloids, steroids, triterpenes, phlobatannins, anthraquinones and glycosides. A 4-day suppressive antimalarial test was carried out using Plasmodium berghei NK65-infected mice, to which the extract was administered at doses of 31.25, 62.5, 125, 250 and 500 mg/kg body weight (BW). Toxicity of the extract was evaluated in rats using selected hematological parameters and organ function indices after orally administering doses of 25, 50 and 100 mg/kg BW for 14 d.

RESULTSPhytochemical analysis revealed the presence of alkaloids, tannins, phenolics, saponins, glycosides, steroids and anthraquinones in the extract. Moreover, the extract reduced parasitemia by 39.2% and 45.8% at doses of 250 and 500 mg/kg BW respectively on day 8 post-inoculation. Various hematological parameters evaluated were not significantly altered (P>0.05) at all doses of the extract, except red blood cell count which was significantly elevated (P<0.05) at 100 mg/kg BW. The extract significantly increased (P<0.05) urea, creatinine, cholesterol, high-density lipoprotein-cholesterol and bilirubin concentrations in the serum as well as atherogenic index, while it reduced albumin concentration significantly (P<0.05) at higher doses compared to the controls. Alanine aminotransferase activity was reduced in the liver and heart significantly (P<0.05) but was increased in the serum significantly (P<0.05) at higher doses of the extract compared to the controls.

CONCLUSIONThe results suggest that methanolic extract of the Dwarf red variety has partial antimalarial activity at higher doses, but is capable of impairing normal kidney and liver function as well as predisposing subjects to cardiovascular diseases.

Animals ; Antimalarials ; administration & dosage ; adverse effects ; pharmacology ; Cocos ; Dose-Response Relationship, Drug ; Malaria ; drug therapy ; Mice ; Plant Extracts ; administration & dosage ; adverse effects ; pharmacology ; Plasmodium berghei ; Rats ; Rats, Wistar

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Effect of Saikokeishito, a Kampo medicine, on hydrogen peroxide-induced premature senescence of normal human dermal fibroblasts.

Takanobu TAKATA ; Yoshiharu MOTOO ; E-mail: MOTOO@KANAZAWA-MED.AC.JP. ; Naohisa TOMOSUGI

Journal of Integrative Medicine.2014;12(6):495-503. doi:10.1016/S2095-4964(14)60052-2

OBJECTIVESaikokeishito (TJ-10) is a Kampo (traditional Japanese herbal) medicine, clinically used for hundreds of years in East Asia. Among its various mechanisms elucidated so far, TJ-10 inhibits the production of transforming growth factor-β1 (TGF-β1) and development of pancreatic fibrosis in vivo. Oxidative damage of normal human dermal fibroblasts (NHDFs) in the corium is a cause of human dermal senescence. Our aim was to determine whether TJ-10 protects NHDFs from premature senescence by hydrogen peroxide (H₂O₂).

METHODSPremature senescence was induced in NHDFs by 200 μmol/L H₂O₂ for 4 h. Cell viability and the expressions of p53, AMP-activated protein kinase α1 (AMPKα1), AMPKα2, and 14-3-3 protein sigma (14-3-3 σ) were measured in NHDFs treated with TJ-10 for 48 h before exposure to H₂O₂for 4 h.

RESULTSCell viability after treatment with 200 μmol/L H₂O₂ for 4 h was similar (about 80%) to after pre-treatment with TJ-10. Ascorbic acid as a control did not protect NHDFs from damage by 200 μmol/L H₂O₂. Treatment with 200 μmol/L H₂O₂tended to up-regulate p53 and to down-regulate SIRT1 and AMPKα1, but had no effect on AMPKα2 and 14-3-3 σ expression. Pretreatment with TJ-10 inhibited H₂O₂-induced up-regulation of p53 and enhanced AMPKα1 expression.

CONCLUSIONIt is suggested that Saikokeishito has a protective effect on oxidative stress-induced senescence of NHDFs.

AMP-Activated Protein Kinases ; metabolism ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cell Culture Techniques ; Cell Survival ; drug effects ; Cellular Senescence ; drug effects ; Dermis ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Medicine, Kampo ; methods ; Sirtuin 1 ; metabolism ; Up-Regulation ; drug effects

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Cytotoxic genes from traditional Chinese medicine inhibit tumor growth both in vitro and in vivo.

Yuan-hui ZHANG ; Yuan WANG ; Ali Hussein YUSUFALI ; Frederick ASHBY ; Daniel ZHANG ; Zi-fei YIN ; George V ASLANIDI ; Arun SRIVASTAVA ; Chang-quan LING ; E-mail: LINGCHANGQUAN@HOTMAIL.COM. ; Chen LING ; E-mail: LINGCHEN@PEDS.UFL.EDU.

Journal of Integrative Medicine.2014;12(6):483-494. doi:10.1016/S2095-4964(14)60057-1

OBJECTIVELittle effort has been made to study the protein-encoding genes isolated from traditional Chinese medicine (TCM) drugs, and the delivery of these genes into malignant cells through recombinant adeno-associated virus (rAAV) vectors has not been attempted.

METHODSWe synthesized the cDNAs of five known cytotoxic proteins isolated from TCM drugs and the FLAG epitope-tagged cDNAs were subcloned into a rAAV plasmid vector. The protein expression was confirmed by Western blot assay. Various cancer cell lines were transfected with the above plasmids and cell growth was monitored both in vitro and in vivo. The best cytotoxic gene was further packaged into rAAV vectors, under the control of a liver cancer-specific promoter. The liver tumor growth was then monitored following intratumor administration of the rAAV vectors.

RESULTSThe expression plasmids, encoding individual potential cytotoxic genes tagged with FLAG epitope, were successfully generated and sequenced. Among these genes, trichosanthin (TCS) gene yielded the most promising results for the inhibition of cancer cell growth in vitro. The over-expressed TCS functioned as a type I ribosome-inactivating protein, followed by inducing apoptosis that is associated with the Bcl-PARP signaling pathway. Furthermore, intratumor injection of rAAV vectors containing the TCS gene significantly inhibited the growth of human hepatocellular carcinoma tumors in a murine xenograft model.

CONCLUSIONOur studies suggest that the use of TCM cytotoxic genes is a useful therapeutic strategy for treating human cancers in general, and liver tumors in particular.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; physiology ; Cell Line, Tumor ; DNA, Complementary ; Dependovirus ; Genetic Vectors ; Humans ; Liver Neoplasms ; physiopathology ; Medicine, Chinese Traditional ; methods ; Mice ; Neoplasms ; physiopathology ; Trichosanthin ; genetics ; pharmacology

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Constructing protein-protein interaction network of hypertension with blood stasis syndrome via digital gene expression sequencing and database mining.

Yong-hong LIAN ; Mei-xia FANG ; Li-guo CHEN ; E-mail: TCHENLY@JNU.EDU.CN.

Journal of Integrative Medicine.2014;12(6):476-482. doi:10.1016/S2095-4964(14)60058-3

OBJECTIVETo construct a protein-protein interaction (PPI) network in hypertension patients with blood-stasis syndrome (BSS) by using digital gene expression (DGE) sequencing and database mining techniques.

METHODSDGE analysis based on the Solexa Genome Analyzer platform was performed on vascular endothelial cells incubated with serum of hypertension patients with BSS. The differentially expressed genes were filtered by comparing the expression levels between the different experimental groups. Then functional categories and enriched pathways of the unique genes for BSS were analyzed using Database for Annotation, Visualization and Integrated Discovery (DAVID) to select those in the enrichment pathways. Interologous Interaction Database (I2D) was used to construct PPI networks with the selected genes for hypertension patients with BSS. The potential candidate genes related to BSS were identified by comparing the number of relationships among genes. Confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), gene ontology (GO) analysis was used to infer the functional annotations of the potential candidate genes for BSS.

RESULTSWith gene enrichment analysis using DAVID, a list of 58 genes was chosen from the unique genes. The selected 58 genes were analyzed using I2D, and a PPI network was constructed. Based on the network analysis results, candidate genes for BSS were identified: DDIT3, JUN, HSPA8, NFIL3, HSPA5, HIST2H2BE, H3F3B, CEBPB, SAT1 and GADD45A. Verified through qRT-PCR and analyzed by GO, the functional annotations of the potential candidate genes were explored.

CONCLUSIONCompared with previous methodologies reported in the literature, the present DGE analysis and data mining method have shown a great improvement in analyzing BSS.

Aged ; Data Mining ; methods ; Databases, Factual ; Female ; Gene Expression ; Hemostatic Disorders ; epidemiology ; genetics ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Protein Interaction Maps

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Role of adult resident renal progenitor cells in tubular repair after acute kidney injury.

Hui-ling WANG ; E-mail: VIOLLLA@163.COM. ; Nan-mei LIU ; Rui LI ;

Journal of Integrative Medicine.2014;12(6):469-475. doi:10.1016/S2095-4964(14)60053-4

Acute kidney injury is a serious global health problem and determinant of morbidity and mortality. Recent advancements in the field of stem cell research raise hopes for stem cell-based regenerative approaches to treat acute kidney diseases. In this review, the authors summarized the latest research advances of the adult resident renal progenitor cells (ARPCs) on kidney repair, the role of ARPCs on tubular regeneration after acute kidney injury, the current understanding of the mechanisms related to ARPC activation and modulation, as well as the challenges that remain to be faced.
Acute Kidney Injury ; physiopathology ; Antigens, CD ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Kidney ; physiopathology ; Kidney Tubules ; physiopathology ; Receptors, CXCR ; metabolism ; Regeneration ; physiology ; Reperfusion Injury ; physiopathology ; Stem Cells ; physiology

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Comparative study on WHO Western Pacific Region and World Federation of Chinese Medicine Societies international standard terminologies on traditional medicine: diseases of Gynecology and Obstetrics (Part 3).

Zhao-Guo LI ; E-mail: ZHOOUSHI@163.COM.

Journal of Integrative Medicine.2014;12(5):463-467. doi:10.1016/S2095-4964(14)60043-1

Gynecology ; Humans ; Medicine, Chinese Traditional ; Obstetrics ; Terminology as Topic ; Translations ; World Health Organization

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Simultaneous determination of five anthraquinones in a Chinese traditional preparation by RP-HPLC using an improved extraction procedure.

Yan-bin SHI ; E-mail: YBYYSH@126.COM. ; Hui-li LI ; Hai-qin WANG ; Yan-biao YANG ; Xiao-yun ZHANG ; Hui WANG ; Zong-jie ZHU ; Zhi-Ye ZHANG ; Cheng-an ZHANG

Journal of Integrative Medicine.2014;12(5):455-462. doi:10.1016/S2095-4964(14)60037-6

OBJECTIVEThe stable quality of Chinese herbal medicines is a critical factor for their reliable clinical efficiency. An improved liquid-liquid extraction procedure and a liquid chromatographic method were developed to simultaneously analyze five anthraquinones (aloe-emodin, rhein, emodin, chrysophanol and physcion) in a Chinese traditional hospital preparation, Fuyankang mixture, in order to quantitatively control its quality in a more effective way.

METHODSA more economical and repeatable extraction procedure based on conventional liquid-liquid extraction technique was developed and used to extract five marker components in Fuyankang mixture. These anthraquinones were separated in less than 20 min on a C18 column with methanol and 0.1% phosphoric acid (88:12, v/v) as mobile phase. The method was validated for specificity, precision, spiked recovery and stability.

RESULTSCompared to conventional liquid-liquid extraction, the improved liquid-liquid extraction was found to be more effective for simultaneous extraction of anthraquinones from an aqueous Chinese herbal preparation, especially for hydrophobic compounds. The improved extraction method was successfully applied to determine the content of five marker components in Fuyankang mixture by the means of reverse phase high-performance liquid chromatography.

CONCLUSIONThe improved extraction procedure may be suitable for routine quality control of Fuyankang mixture and other traditional preparations at city-level hospitals in China.

Anthraquinones ; analysis ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; analysis ; Emodin ; analogs & derivatives ; analysis

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Liuwei Dihuang, a traditional Chinese herbal formula, suppresses chronic inflammation and oxidative stress in obese rats.

Benjamin PERRY ; Junzeng ZHANG ; Tarek SALEH ; Yanwen WANG ; E-mail: YANWEN.WANG@NRC.CA.

Journal of Integrative Medicine.2014;12(5):447-454. doi:10.1016/S2095-4964(14)60044-3

OBJECTIVETo investigate the anti-inflammatory, anti-oxidative stress, and adipokine-ameliorating effects of Liuwei Dihuang (LWDH), a traditional Chinese herbal formula, in obese rats.

METHODSAfter 2 weeks of acclimation with free access to regular rodent chow and water, obese-prone-caesarean-derived (OP-CD) rats were fed a modified AIN-93G diet containing 60% energy from fat. Treatment was performed twice daily by gavage feeding with 500, 1 500, or 3 500 mg/kg body weight LWDH suspended in water (n=12 rats per group). Twelve obese-resistant-CD (OR-CD) rats were fed the atherogenic diet and gavaged with water, and served as the normal control. Blood biomarkers of inflammation, oxidative stress and adiponectin were measured post-sacrifice and used to determine the treatment effect of LWDH and assess the suitability of OR/OP-CD rats for studying these parameters.

RESULTSAfter 9 weeks of treatment, LWDH lowered serum C-reactive protein (CRP) and tumour necrosis factor-α (TNF-α) levels. Serum interleukin-6 (IL-6) levels showed a tendency towards reduction, but were not significantly different from the OP-CD control. Liver superoxide dismutase (SOD) activity was increased in response to all three doses of LWDH, while the levels of reduced (GSH) and oxidized glutathione (GSSG) and thiobarbituric acid reactive substances (TBARS) were unchanged. Serum adiponectin levels were increased in response to oral administration of LWDH at the dose of either 500 or 1 500 mg/kg body weight. In addition, comparisons between OR-CD and OP-CD rats revealed differential, and for some biomarkers, conflicting characteristics of high-fat diet-fed OP-CD rats in reference to obese human subjects in terms of inflammatory and oxidative stress biomarkers and circulating adiponectin levels.

CONCLUSIONThe results show, for the first time, the anti-inflammatory, anti-oxidative stress and adiponectin-ameliorating effects of LWDH in obese rats. The suitability of the OR/OP-CD rat model as a research tool to study inflammation, oxidative stress, and adipokine production requires further investigation.

Adiponectin ; blood ; Animals ; C-Reactive Protein ; analysis ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Glutathione ; metabolism ; Inflammation ; drug therapy ; Interleukin-6 ; blood ; Male ; Obesity ; complications ; Oxidative Stress ; drug effects ; Rats

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Effect of aqueous extract of Solanum xanthocarpum Schrad. & Wendl. on postmenopausal syndrome in ovariectomized rats.

Urmila ASWAR ; Email: ASWARURMILA@GMAIL.COM. ; Mayuri GURAV ; Ganesh MORE ; Khaled RASHED ; Manoj ASWAR

Journal of Integrative Medicine.2014;12(5):439-446. doi:10.1016/S2095-4964(14)60042-X

OBJECTIVESolanum xanthocarpum Schrad. & Wendl. (Solanaceae) is present in many Ayurveda compound formulations including Chavanaprasha and Dasamoolarishta. The whole plant is used in conditions such as inflammation, constipation and promoting conception in females. In the present study, we carried out different tests to evaluate the effect of aqueous extract of Solanum xanthocarpum (SXE) in postmenopausal syndrome.

METHODSThe study was carried out in bilaterally ovariectomized one-month-old Wistar rats (40-50 g). Bilaterally ovariectomized (OVX) Wistar rats were divided into four groups (n=6) receiving different treatments, consisting of a vehicle (distilled water), aqueous extract of Solanum xanthocarpum at two different doses (200 and 400 mg/kg) administered orally daily for 90 d and standard drug β estradiol at a dose of 1 mg/kg administered subcutaneously biweekly for 90 d. Estrogenic activity was assessed by vaginal cornification, sexual behavior, serum estradiol and uterine weight to body weight ratio. Antiosteoporotic activity was assessed on the basis of biomechanical and biochemical parameters followed by histopathological studies, and antidepressant activity was assessed by forced swim test.

RESULTSSXE showed presence of steroids. At the dose of 200 mg/kg, it significantly improved all the parameters of sexual behavior (P<0.01), caused vaginal cornification, and increased serum estradiol and uterine weight (P<0.01). It also significantly improved all the parameters of bone strength as well as depression (P<0.01). Histopathology of bones confirmed the above findings.

CONCLUSIONThe study indicated that SXE may provide an effective treatment in the prevention of postmenopausal symptoms.

Animals ; Antidepressive Agents ; pharmacology ; Estradiol ; blood ; Estrogens ; pharmacology ; Female ; Femur ; drug effects ; metabolism ; Humans ; Male ; Mice ; Osteoporosis, Postmenopausal ; prevention & control ; Ovariectomy ; Plant Extracts ; pharmacology ; Postmenopause ; drug effects ; Rats ; Rats, Wistar ; Sexual Behavior, Animal ; drug effects ; Solanum

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Low doses of ethanolic extract of Boldo (Peumus boldus) can ameliorate toxicity generated by cisplatin in normal liver cells of mice in vivo and in WRL-68 cells in vitro, but not in cancer cells in vivo or in vitro.

Jesmin MONDAL ; Kausik BISHAYEE ; Ashis Kumar PANIGRAHI ; Anisur Rahman KHUDA-BUKHSH ; E-mail: prof_arkb@yahoo.co.in, KHUDABUKHSH_48@REDIFFMAIL.COM.

Journal of Integrative Medicine.2014;12(5):425-438. doi:10.1016/S2095-4964(14)60045-5

OBJECTIVEUse of cisplatin, a conventional anticancer drug, is restricted because it generates strong hepatotoxicity by accumulating in liver. Therefore its anticancer potential can only be fully exploited if its own toxicity is considerably reduced. Towards this goal, ethanolic extract of the plant, Boldo (Peumus boldus), known for its antihepatotoxic effects, was used simultaneously with cisplatin, to test its ability to reduce cisplatin's cytotoxicity without affecting its anticancer potential.

METHODSThe cytotoxicity of Boldo extract (BE) and cisplatin, administered alone and in combination, was determined in three cancer cell lines (A549, HeLa, and HepG2) and in normal liver cells (WRL-68). Drug-DNA interaction, DNA damage, cell cycle, apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP, ΔΨ) were also studied. Hepatotoxicity and antioxidant activity levels were determined by alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and glutathione assays in mice. The cytotoxicity of related proteins was tested by Western blotting.

RESULTSCo-administration of BE and cisplatin increased viability of normal cells, but had no effect on the viability of cancer cells. Boldo protected liver from damage and normalized different antioxidant enzyme levels in vivo and also reduced ROS and re-polarized MMP in vitro. Bax and cytochrome c translocation was reduced with caspase 3 down-regulation. Further, a drug-DNA interaction study revealed that BE reduced cisplatin's DNA-binding capacity, resulting in a reduction in DNA damage.

CONCLUSIONResults indicated that a low dose of BE could be used beneficially in combination with cisplatin to reduce its toxicity without hampering cisplatin's anticancer effect. These findings signify a potential future use of BE in cancer therapy.

Animals ; Antineoplastic Agents ; toxicity ; Cells, Cultured ; Cisplatin ; toxicity ; DNA Damage ; Female ; Glutathione ; metabolism ; Hepatocytes ; drug effects ; metabolism ; pathology ; Humans ; Male ; Mice ; Neoplasms ; drug therapy ; pathology ; Peumus ; Plant Extracts ; pharmacology

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