Design and synthesis of 6,20-epoxy A-ring modified oridonin derivatives with antitumor activity through extrinsic and mitochondrial pathways.

Oridonin is an antitumor ent-kaurane diterpenoid that medicinal chemists have been paying close attention to in recent years. Herein, a novel 6,20-epoxy A-ring modified oridonin derivative 2 was obtained by a 6-step synthesis. A series of 14-O derivatives of 2 (EpskA1-EpskA24) were synthesized to further enhance the activity. Based on their cytotoxicity against MCF-7, A549 and L-02 cells, EpskA9, EpskA10 and EpskA21 were chosen for further screening to obtain a wider antitumor spectrum. Collectively, EpskA21 showed the most potent antiproliferative activity, inhibiting proliferation and migration, and inducing apoptosis and cell cycle arrest in MCF-7 and MIA-PaCa-2 cells. With the help of network pharmacology analysis, apoptosis-related proteins were selected and further tested by western blot assay. The inhibition of PI3K/AKT and an increase in the levels of Bax/Bcl-2 ratio, Cyt-C, cleaved-Caspase-9, cleaved-Caspase-3 and cleaved-PARP was observed, indicating that EpskA21 induced apoptosis through the mitochondrial pathway. Given that an increase in DR5 expression and activated Caspase-8 were also observed, the extrinsic apoptosis pathway might also be related to the antitumor effect.

Antidepressant-like effects of Cimicifuga dahurica (Turcz.) Maxim. via modulation of monoamine regulatory pathways.

Sheng-ma is recorded in the Compendium of Materia Medica and mainly originates from the rhizomes of Cimicifuga dahurica (Turcz.) Maxim. (CD), Cimicifuga heracleifolia Kom. and Cimicifuga foetida L. The alcoholic extract of Cimicifuga foetida L. (Brand name: Ximingting®) has been approved for the treatment of perimenopausal symptoms accompanying hot flash, depression and anxiety in China. However, there's no further study about the antidepressant-like effects of C. dahurica (CD). The aim of this study is to investigate the antidepressant-like effect of CD extracted by 75% ethanol and its possible mechanisms.The neuro-protective effects of CD on injured PC12 cells induced by corticosterone was measured firstly. Then, forced swim test (FST), tail suspension test (TST), reserpine-induced hypothermia, 5-hydroxytryptophan (5-HTP) induced head twitch response in mice and chronic unpredictable mild stress (CUMS) on sucrose preference tests were executed. Moreover, the potential mechanisms were explored by measuring levels of monoamine neurotransmitter in mice frontal cortex and hippocampus, testing monoamine oxidase enzyme A (MAO-A) activities in the brains of CUMS-exposed mice. Results showed that CD (60, 120 mg/kg) can significantly decreased the immobility period in FST and TST in mice without affecting locomotor activity. CD (30 mg/kg, 60 mg/kg, 120 mg/kg) could significantly counteracted reserpine-induced hypothermia and increased the number of head-twitches in 5-HTP induced head twitch response. It was also found that the monoamine neurotransmitter levels in the hippocampus and frontal cortex were significantly increased in 60 mg/kg and 120 mg/kg CD treated mice. In addition, CD (60 and 120 mg/kg) significantly inhibited MAO-A after 6-week CUMS exposure. CD can effectively produce an antidepressant-like effect, which involved with modulation of monoamine regulatory pathways.

Three new phenols and one new lignan from Clematis terniflora var. manshurica (Rupr.) Ohwi with their anti-inflammatory activity.

Three undescribed phenols, mandshusica C-E (1-3) and a new lignan, mandshusica F (5), along with six known compounds (4, 6-10) were isolated from the roots and rhizomes of Clematis terniflora var. manshurica (Rupr.) Ohwi. Their structures were elucidated by extensive spectroscopic analysis as well as NMR and ECD calculations. Moreover, the possible biosynthetic pathways of compounds 1-3 were also discussed. All compounds were evaluated for their anti-inflammatory activities in LPS-induced RAW 264.7 cells. Compounds 1, 3, 4 significantly reduced the levels of NO and TNF-α, while compounds 2 and 8 significantly inhibited NO production in LPS-induced RAW264.7 cells.

Mono-ADP-ribosylation, a MARylationmultifaced modification of protein, DNA and RNA: characterizations, functions and mechanisms.

The functional alterations of proteins and nucleic acids mainly rely on their modifications. ADP-ribosylation is a NAD+-dependent modification of proteins and, in some cases, of nucleic acids. This modification is broadly categorized as Mono(ADP-ribosyl)ation (MARylation) or poly(ADP-ribosyl)ation (PARylation). MARylation catalyzed by mono(ADP-ribosyl) transferases (MARTs) is more common in cells and the number of MARTs is much larger than poly(ADP-ribosyl) transferases. Unlike PARylation is well-characterized, research on MARylation is at the starting stage. However, growing evidence demonstrate the cellular functions of MARylation, supporting its potential roles in human health and diseases. In this review, we outlined MARylation-associated proteins including MARTs, the ADP-ribosyl hydrolyses and ADP-ribose binding domains. We summarized up-to-date findings about MARylation onto newly identified substrates including protein, DNA and RNA, and focused on the functions of these reactions in pathophysiological conditions as well as speculated the potential mechanisms. Furthermore, new strategies of MARylation detection and the current state of MARTs inhibitors were discussed. We also provided an outlook for future study, aiming to revealing the unknown biological properties of MARylation and its relevant mechanisms, and establish a novel therapeutic perspective in human diseases.

Production improvement of an antioxidant in cariogenic Streptococcus mutans UA140.

AIMS: This study aimed to improve the production of mutantioxidin, an antioxidant encoded by a biosynthetic gene cluster (mao) in Streptococcus mutans UA140, through a series of optimization methods. METHOD AND RESULTS: Through the construction of mao knockout strain S. mutans UA140∆mao, we identified mutantioxidin as the antioxidant encoded by mao and verified its antioxidant activity through a reactive oxygen species (ROS) tolerance assay. By optimizing the culture medium and fermentation time, 72 h of fermentation in chemically defined medium (CDM) medium was determined as the optimal fermentation conditions. Based on two promoters commonly used in Streptococcus (ldhp and xylS1p), eight promoter refactoring strains were constructed, nevertheless all showed impaired antioxidant production. In-frame deletion and complementation experiments demonstrated the positive regulatory role of mao1 and mao2, on mao. Afterward, the mao1 and mao2, overexpression strain S. mutans UA140/pDL278:: mao1mao2, were constructed, in which the production of mutantioxidin was improved significantly. CONCLUSIONS: In this study, through a combination of varied strategies such as optimization of fermentation conditions and overexpression of regulatory genes, production of mutantioxidin was increased by 10.5 times ultimately.

Two new prenylated flavonol glycosides from Epimedium koreanum Nakai leaves and their anti-pulmonary fibrosis activities.

Two new prenylated flavonol glycosides, namely Desmethylicaritin-3-O-ß-D-glucopyranosyl-(1→3)-α-L(4″-O-acetyl) rhamnopyranosyl-7-O-ß-D(6''''-O-acetyl)-glucopyranoside (1) and 5,7,3',4'-tetrahydroxyl-8-(3,3-dimethylallyl)-flavonol-3-O-α-L-rhamnopyranoside (2), and one with no NMR spectral data reported (3) were isolated from Epimedium koreanum Nakai. Their structures were elucidated by 1D, 2D NMR and HRESIMS analysis. The identification of the sugar moieties was carried out by means of acid hydrolysis and HPLC analysis of their derivatives. The anti-pulmonary fibrosis activities result showed that compound 2 had significant inhibitory effects on A549 cell fibrosis, which was similar to that of the positive control drug, pirfenidone.

Sodium nitroprusside improved the quality of Radix Saposhnikoviae through constructed physiological response under ecological stress.

The ecological significance of secondary metabolites is to improve the adaptive ability of plants. Secondary metabolites, usually medicinal ingredients, are triggered by unsuitable environment, thus the quality of medicinal materials under adversity being better. The quality of the cultivated was heavily declined due to its good conditions. Radix Saposhnikoviae, the dried root of Saposhnikovia divaricata (Turcz.) Schischk., is one of the most common botanicals in Asian countries, now basically comes from cultivation, resulting in the market price being only 1/10 to 1/3 of its wild counterpart, so improving the quality of cultivated Radix Saposhnikoviae is of urgency. Nitric oxide (NO) plays a crucial role in generating reactive oxygen species and modifying the secondary metabolism of plants. This study aims to enhance the quality of cultivated Radix Saposhnikoviae by supplementing exogenous NO. To achieve this, sodium nitroprusside (SNP) was utilized as an NO provider and applied to fresh roots of S. divaricata at concentrations of 0.03, 0.1, 0.5, and 1.0 mmol/L. This study measured parameters including the activities of antioxidant enzymes, secondary metabolite synthesis enzymes such as phenylalanine ammonia-lyase (PAL), 1-aminocyclopropane-1-carboxylic acid (ACC), and chalcone synthase (CHS), as well as the contents of NO, superoxide radicals (O2·-), hydrogen peroxide (H2O2), malondialdehyde (MDA), and four secondary metabolites. The quality of Radix Saposhnikoviae was evaluated with antipyretic, analgesic, anti-inflammatory effects, and inflammatory factors. As a result, the NO contents in the fresh roots were significantly increased under SNP, which led to a significant increase of O2·-, H2O2, and MDA. The activities of important antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), were found to increase as well, with their peak levels observed on the 2nd and 3rd days. PAL, ACC, and CHS activities were also significantly enhanced, resulting in the increased secondary metabolite contents of Radix saposhnikoviae in all groups, especially the 0.5 mmol/L SNP. The four active ingredients, prim-O-glucosylcimifugin, cimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, and sec-O-glucosylhamaudol, increased by 88.3%,325.0%, 55.4%, and 283.8%, respectively, on the 3rd day. The pharmaceutical effects of Radix Saposhnikoviae under 0.5 mmol/L SNP were significantly enhanced. Exogenous SNP can induce the physiological response of S. divaricata under adverse conditions and significantly improve the quality of Radix Saposhnikoviae.

Two new anti-inflammatory compounds from the roots and rhizomes of Clematis terniflora var. manshurica (Rupr.) Ohwi.

Two new compounds named mandshurica A (1) and mandshurica B (2), together with four known lignans (3-6) were isolated from the roots and rhizomes of Clematis terniflora var. manshurica (Rupr.) Ohwi. The structures of the new compounds were elucidated by HR-ESI-MS, 1D and 2D NMR spectroscopy. Moreover, the anti-inflammatory activity of compounds 1 and 2 were evaluated against lipopolysaccharide-induced mouse macrophage RAW264.7 cells. Compounds 1 and 2 displayed significant inhibitory effect on NO production, and compound 2 exhibited obvious inhibition on the pro-inflammatory cytokines TNF-α. Both new compounds showed potential anti-inflammatory activity.

Ginsenoside Rb1 for overcoming cisplatin-insensitivity of A549/DDP cells in vitro and vivo through the dual-inhibition on two efflux pumps of ABCB1 and PTCH1.

BACKGROUND: The multi-drug resistance is an inherent weakness in the chemotherapeutics of non-small cell lung cancer occurring frequently all over the world. Clinically, ginseng and Chinese medicinal prescriptions including ginseng usually used as anti-tumor adjuncts due to its characteristic of qi-invigorating, which could improve the curative effect of chemotherapy drugs and reduce their toxic side effects. Triterpenoid saponins are the crucial active ingredients in Panax ginseng, and Ginsenoside Rb1 is of the highest quantities. However, the research on the tumor drug-resistance reversal effect and mechanism of ginsenoside Rb1 is still not clear. PURPOSE: This study aimed to systematically estimate the reversal activity of Ginsenoside Rb1 on cisplatin-insensitivity of A549/DDP cells and to reveal its prospective molecular mechanism. METHODS: MTT assay were conducted to evaluate the reversal activity on cisplatin-insensitivity of A549/DDP cells of Ginsenoside Rb1in vitro, and the behavior was also studied by establishing a subcutaneous transplanted tumor model of A549/DDP in BALB/c-nu mice. In addition, P-gp ATPase activity assay, cisplatin accumulation assay, Annexin V-FITC apoptosis assay, real-time qPCR analysis and western blotting analysis were used to clarify the potential mechanism. RESULTS: Ginsenoside Rb1 could effectively reverse the cisplatin-resistance of A549/DDP in vitro and vivo. And after the co-treatment of Ginsenoside Rb1 plus cisplatin, the accumulation of cisplatin increased in A549/DDP cells, which was accompanied with the down-regulation of the mRNA and protein expression levels of ABCB1, SHH, PTCH1 and GLI2. Besides, the apoptosis-inducing ability of cisplatin improved by the relative regulation on the protein expression level of Bax and Bcl-2. Far more importantly, the changes of CYP3A4 mRNA and protein levels were not significant. CONCLUSION: Ginsenoside Rb1 could increase the concentration of intracellular cisplatin and improve the insensitivity for cisplatin on A549/DDP cells. Even better, there was perhaps no unpredictable CYP3A4-mediated pharmacokinetic interactions after the combination of Ginsenoside Rb1 plus cisplatin. Ginsenoside Rb1 was a probable reversal agent for the cisplatin-insensitivity of A549/DDP cells, with a bifunction of inhibiting the efflux of two drug pumps (P-gp and PTCH1) by targeting ABCB1 and Hedgehog (Hh) pathway. In general, this research laid the groundwork for the development of a new reversal agent for the cisplatin-insensitivity of NSCLC.

Icariin, the main prenylflavonoid of Epimedii Folium, ameliorated chronic kidney disease by modulating energy metabolism via AMPK activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedii Folium is a famous traditional Chinese medicine (TCM) widely used in classic formulas, Chinese patent drugs and health care products for treating kidney diseases. Therefore, we speculated that icariin, its main component, might also have a good therapeutic effect on chronic kidney disease (CKD). AIM OF STUDY: To investigate the efficacy and potential mechanism of icariin on CKD. MATERIALS AND METHODS: A CKD model was established by intragastric administration of adenine (200 mg/kg/d) to adult male SD rats for 28 consecutive days. TGF-ß1-induced fibrotic HK-2 cells were applied to establish the renal fibrosis model in vitro. Biochemical determination, pathological staining, flow cytometry and ELISA were performed to preliminarily evaluate the renoprotection of icariin. The intervention effect of icariin on renal fibrosis progression was assessed by cell stiffness determination and multiple immunological methods. The potential mechanism of icariin on CKD was revealed by means of 1H NMR metabolomics, qRT-PCR and Western blotting analysis. RESULTS: Icariin at the dosage of 100 mg/kg/d and 200 mg/kg/d markedly ameliorated rat renal function in a dose-dependent manner. Based on renal pathological features, the mechanism of icariin intervention in CKD was initially revealed by metabolomics, which was closely related to energy metabolism pathways. Furthermore, the detection results of AMPK and related factors in its mediated signaling pathways indicated that icariin exerted a therapeutic effect on CKD by attenuating inflammation and oxidative stress responses and retarding renal fibrosis progression through regulating AMPK/SIRT1/NF-κB and AMPK/ACC signaling pathways. CONCLUSION: It was the first time to demonstrate that icariin could treat adenine-induced CKD by modulating energy metabolism via AMPK activation in a dose-dependent manner.

Four new compounds isolated from the rhizomes of Phedimus aizoon (L.) 't Hart.

One new coumarin, one new flavonoid, two new distinctive compounds with characteristics of an iriflophene unit and a flavonoid unit connecting via a furan ring, together with five known compounds were isolated from the rhizomes of Phedimus aizoon (L.) 't Hart. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 4, 5 and 8 increased the survival rate of H9c2 cells induced by doxorubicin in vitro, which was capable of further drug exploration.

Two new dammarane-type ginsenosides from Panax ginseng.

Two new dammarane-type ginsenosides elucidated as 3-O-[ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl-(1→2)-ß-D-glucopyranosyl]-20-O-[α-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl]-20(S)-protopanaxadiol, named Ginsenoside Rb6(2); (Z)-6-O-ß-D-glucopyranosyl-dammar-20(22), 24-diene-3ß, 6α, 12ß-triol, named (20Z)-Ginsenoside Rh4 (4); along with five known ginsenosides(1, 3, 5, 6, 7) were isolated from the radix and rhizome of wild ginseng (Panax ginseng). The structures were determined by chemical and comprehensive spectroscopic analysis, including 1D, 2D NMR, HR-ESI-MS and hydrolysis reaction.

Systematic analysis of the material basis and mechanism of total saponins of mountain cultivated ginseng against doxorubicin-induced cardiotoxicity based on integrating network pharmacology and in vivo substance profiling.

INTRODUCTION: Doxorubicin-induced cardiotoxicity (DIC) is a serious obstacle to oncologic treatment. Mountain cultivated ginseng (MCG) exhibits stronger pharmacological effects than cultivated ginseng (CG) mainly due to the differences in ginsenosides. However, the material basis and the underlying mechanism of the protective effects of total saponins of MCG (TSMCG) against DIC are unclear. OBJECTIVES: We aimed to elucidate the material basis and the pharmacodynamic effects of TSMCG on DIC as well as the underlying mechanisms. METHODS: To comprehensively analyze the effective substances, the chemical components of TSMCG and their prototypes or metabolites in vivo were characterized through UHPLC/Q-TOF-MS. Then, an absorbed component-target-disease network was established to explore the mechanisms underlying the protective effects of TSMCG against DIC. H9c2 cells were employed for pharmacodynamic assays. The mechanism was verified by Western blot and molecular docking simulations. RESULTS: A total of 56 main ginsenosides were identified in TSMCG, including 27 ginsenosides of PPD type, 15 ginsenosides of PPT type, two ginsenosides of OA types, and 12 ginsenosides of other types. Moreover, 55 ginsenoside prototypes or metabolites in vivo were tentatively characterized. Ginsenoside Ra1 , a differential compound between MCG and CG, could be metabolized by oxidation and deglycosylation. Network pharmacology showed that AKT1, p53, and STAT3 are core targets of 62 intersecting genes. Molecular docking results indicated that most of the ginsenosides have favorable affinity with these core targets. After doxorubicin exposure, TSMCG could increase cell viability and inhibit apoptosis in a dose-dependent manner. CONCLUSION: Our work reveals a novel comprehensive strategy to study the material basis of the protective effects of TSMCG against DIC and the underlying mechanisms through integrating in vivo substance identification, metabolic profiling, network pharmacology, pharmacodynamic evaluation, and mechanism verification.

Structure characterization and anti-fatigue activity of an acidic polysaccharide from Panax ginseng C. A. Meyer.

ETHNOPHARMACOLOGICAL RELEVANCE: Panax ginseng C. A. Meyer is a traditional Chinese herbal medicine, which has been used in China for more than 2000 years. Its traditional effect of "invigorating vitality" is mainly reflected in anti-fatigue. However, due to the difficulty of identification of polysaccharide structure, there are few reports on homogeneous ginseng polysaccharide, and the molecular mechanism of its anti-fatigue effect remains to be further explored. AIM OF THE STUDY: In order to find the homogenous ginseng polysaccharide with the most anti-fatigue effect, this study is for the first time extracted, isolated and structurally identified polysaccharide monomer from Mountain Cultivated Ginseng (MCG). Then the anti-fatigue activity and molecular mechanism were studied. MATERIALS AND METHODS: The structure of ginseng acidic polysaccharide APS-1 prepared by high performance gel permeation chromatography (HPGPC) was determined by acid hydrolysis/HPLC, methylation/GC-MS and NMR analysis. Anti-fatigue effect was evaluated by exhaustive swimming model, and AMPK axis-related proteins were detected by Western blot. RESULTS: APS-1 significantly prolonged fatigue tolerance time, alleviated accumulation of BLA, LDH and BUN, increased activities of SOD and CAT, alleviated oxidative damage caused by MDA, increased activity of CK, regulated glycolysis, and alleviated muscle fiber contraction. The expressions of LKB1, p-AMPK, PGC-1α and Glut4 in muscle were significantly up-regulated. CONCLUSIONS: The anti-fatigue effect of APS-1 was significantly, and the molecular mechanism may be related to the activation of AMPK axis signaling pathway to improve glucose uptake and mitochondrial function.

Two new dammarane-type saponins from radix and rhizomes of Panax ginseng C. A. Meyer.

Two new dammarane-type ginsenosides elucidated as 6-O-[α-D-glucopyranosyl-(1→3)-ß-D-glucopyranosyl]-dammar-24-ene-3ß, 6α, 12ß, 20S-tetraol, named 20(S)-Ginsenoside Re10 (4); 6-O-[α-L-rhamnopyranosyl-(1→2)-ß-D-glucopyranosyl]-20-O-[α-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl]-dammar-24-ene-3ß, 6α, 12ß, 20R-tetraol, named 20(R)-Ginsenoside Re11 (8); along with one steroidal saponin (1) and six known triterpenoid ginsenosides (2, 3, 5, 6, 7 and 9) were isolated from the radix and rhizomes of mountain-cultivated ginseng (Panax ginseng C. A. Meyer, family Araliaceae). Their structures were determined by comprehensive chemical and spectroscopic analysis. In addition, what's even more concerning is that protodioscin was isolated for the first time from Panax ginseng.

A glycoprotein from mountain cultivated ginseng: Insights into their chemical characteristics and intracellular antioxidant activity.

A glycoprotein (MGP2) from mountain-cultivated ginseng (MCG) was purified by Tris-HCl extraction followed by DEAE-52 ion exchange chromatography and Sephadex G-100 gel filtration chromatography. The approximate molecular weight (27.0 kDa) and monomeric nature were determined by reduced and non-reduced SDS-PAGE. The structure of MGP2 was characterized by a practical and reliable "protein-polysaccharide analyzed by spectroscopy combined with chemical analysis" strategy. The results showed that MGP2 belonged to Arabinogalactan proteins (AGPs) which contained high amount of Glc (35.1 %). The hemagglutination test concluded that MGP2 was not a lectin. In addition, the MGP2 exhibited antioxidant activity by scavenging radical capacity tests and the ability to protect human erythrocytes and RAW264.7 cells from oxidative damage induced by AAPH. Therefore, these results suggested that glycoprotein MGP2 could be used as a natural antioxidant in drug and food industry.

New anti-pulmonary fibrosis prenylflavonoid glycosides from Epimedium koreanum.

Four new prenylflavonoid glycosides, namely koreanoside H-K (1-4), together with eleven known ones (5-15) were isolated from the leaves of Epimedium koreanum Nakai. Their structures were elucidated by 1D NMR, 2D NMR, HR-ESI-MS, IR and UV. The identification of the sugar moieties was carried out by means of acid hydrolysis and HPLC analysis of their derivatives. It is worth noting that compound 3 and compound 4 were elucidated to contain fucose and quinovose moieties, which were two extremely rare sugar units from the genus Epimedium. The anti-pulmonary fibrosis activity of the new compounds was evaluated using A549 cell line. Compounds 1, 2 and 4 showed significant anti-pulmonary fibrosis activities.

Fibrous Roots of Cimicifuga Are at Risk of Hepatotoxicity.

The cause of liver damage by using black cohosh preparation has been concerned but remains unclear. After a preliminary investigation, the black cohosh medicinal materials sold in the market were adulterated with Asian cohosh (Cimicifuga) without removing the fibrous roots. The safety of Cimicifuga rhizome and fibrous roots is unknown and has not been reported. Therefore, in this paper, the rhizome and fibrous roots of Cimicifuga dahurica (Turcz.) Maxim (C. dahurica) were completely separated, extracted with 70% ethanol, and freeze-dried to obtain crude rhizome extract (RC) and fibrous roots extract (FRC). UHPLC-Q-TOF-MS was used to identify 39 compounds in the rhizome and fibrous roots of Cimicifuga, mainly saponins and phenolic acids. In the L-02 cytotoxicity experiment, the IC50 of fibrous roots (1.26 mg/mL) was slightly lower than that of rhizomes (1.417 mg/mL). In the 90-day sub-chronic toxicity study, the FRC group significantly increased the level of white blood cells, ALP, ALT, AST, BILI and CHOL (p < 0.05); large area of granular degeneration and balloon degeneration occurred in liver tissue; and the expression of p-NF-kB in the nucleus increased in a dose-dependent manner. Overall, Fibrous roots of Cimicifuga are at risk of hepatotoxicity and should be strictly controlled and removed during the processing.

Total flavonoids in Epimedium koreanum Nakai alleviated chronic renal failure via promoting AMPK activation.

Chronic renal failure (CRF) is a result of the progression of chronic kidney diseases (CKD), a global health problem with a high cost of treatment and no ideal therapy. The aim of this study is to evaluate the pharmacological efficacy of the total flavonoids in Epimedium koreanum Nakai (TFE), a dietary supplement, against CRF and to determine the mechanism of actions. An adenine-induced CRF rat model and a TGF-ß1 induced human kidney proximal tubule epithelial (HK-2) cell based in vitro renal fibrosis model were established and used to evaluate TFE's efficacy. Renal hemodynamics, biochemical indexes, inflammatory cytokines, histopathology and the reactive oxygen species (ROS) levels were determined to evaluate the efficacy of TFE on CRF. NMR-based metabolomics, immunohistochemical (IHC) staining, immunofluorescence (IF) staining, quantitative real time-PCR (qRT-PCR) and western blotting were conducted to determine the mechanism. The results showed that TFE had a significant effect on CRF at 150 mg kg-1 d-1 and could significantly alleviate renal fibrosis in the animal model. Twelve potential biomarkers, which mainly involve energy metabolism pathways, for CRF were identified using the metabolomics approach. The mechanism study suggested that TFE regulated AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) and AMPK/silent information regulator 1 (SIRT1)/nuclear factor kappa-B (NF-κB) signaling pathways. Furthermore, the effect of TFE was inhibited by compound C in the in vitro experiment, which also confirmed the above conclusion.

Isolation and structural elucidation of a low-molecular-weight polysaccharide from the roots of Panax ginseng C. A. Meyer.

A low-molecular-weight polysaccharide named MCGP-L was extracted and purified from the roots of Mountain cultivated ginseng (Panax ginseng C. A. Meyer). The polysaccharide MCGP-L was purified by molecular exclusion chromatography using the Sephadex G-25 column. The average molecular weight of MCGP-L was estimated to be 3 × 103 kDa. Monosaccharide composition analysis showed MCGP-L was composed of three kinds of monosaccharide: D-glucose, D-galactose and D-mannose. The physicochemical properties and structural characteristics of MCGP-L were investigated by the combination of chemical and instrumental analysis such as methylation analysis, High Performance Gel-Permeation Chromatography (HPGPC), High Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR). The backbone of MCGP-L was composed of (1→4)-linked-α-D-Glcp residues and with branch chain substituted at O-6 position of (1→4,6)-linked-α-D-Glcp. The branch chain consists of →6)-α-D-Galp-(1→, →2)-α-D-Manp-(1→ and ß-D-Glcp-(1→.