Lactobacillus plantarum TWK10-fermented soymilk improves cognitive function in type 2 diabetic rats.

BACKGROUND: The brain is especially sensitive to diabetes-induced damage. Chronic hyperglycemia can potentially lead to brain dysfunctions, affecting spatial learning and memory. RESULTS: The type 2 diabetes (T2D) rats were administered TWK10-fermented soy milk water extract (WE) and ethanol extract (EE) for 6 weeks. WE and EE treatment attenuated T2D-induced alteration in cognitive function assessed using the Morris water maze. Moreover, administration of WE and EE significantly elevated superoxide dismutase activity (166.96% and 181.21%, P < 0.05, respectively) and reduced malondialdehyde concentration (35.03% and 43.97%, P < 0.05, respectively) in the hippocampus of the rats. Additionally, the calmodulin level and nitric oxide concentration were regulated by WE and EE. CONCLUSION: This study provides scientific evidence that WE and EE enhance anti-oxidative enzyme activity, which subsequently regulates factors associated with cognitive function in T2D rats. © 2020 Society of Chemical Industry.

Effect of ethanol extract from Lactobacillus plantarum TWK10-fermented soymilk on wound healing in streptozotocin-induced diabetic rat.

Wound healing is a highly dynamic phenomenon comprising numerous coordinated steps including homeostasis/coagulation, inflammation, migration, proliferation, and remodeling. Diabetes mellitus (DM) is a multisystem chronic epidemic that prolongs inflammation in wounds and is associated with impaired healing. This study aimed to investigate the effect of an ethanol extract from Lactobacillus plantarum TWK10 (TWK10)-fermented soymilk on wound healing. The anti-inflammatory effects of the ethanol extract of TWK10-fermented soymilk on lipopolysaccharide-stimulated RAW264.7 macrophage cells were examined. The ethanol extract of TWK10-fermented soymilk (100 µg/mL) significantly decreased nitric oxide production from 11.34 ± 0.74 µM to 8.24 ± 2.02 µM (p < 0.05) and enhanced proliferation in Detroit 551 cells cultured in high-glucose medium; the cell number peaked at 128.44 ± 7.67% (compared to the untreated control) at 600 µg/mL. An ethanol extract of TWK10-fermented soymilk + vaseline-treated rat model of streptozotocin-induced diabetic wounds was generated herein, and the following groups were formed herein: normal control (NC), blank control (BC), low dose group (LD, 0.24 mg/wound), intermediate dose (MD, 0.48 mg/wound), and high dose (HD, 2.40 mg/wound). On day 14 after wound infliction, the wound area in the LD, MD, and HD groups was significantly decreased to 10.2, 8.4, and 8.5% respectively (p < 0.05). Moreover, in the LD, MD, and, HD groups, tumor necrosis factor-α, interleukin 6, and matrix metalloproteinase-9 were downregulated in the wounded skin. These results show that the topical application of the ethanol extract of TWK10-fermented soymilk is beneficial for enhancing wound healing and for the closure of diabetic wounds.

Glyceryl 1,3-Dipalmitate Produced from Lactobacillus paracasei subspecies. paracasei NTU 101 Inhibits Oxygen-Glucose Deprivation and Reperfusion-Induced Oxidative Stress via Upregulation of Peroxisome Proliferator-Activated Receptor γ in Neuronal SH-SY5Y Cells.

Glyceryl 1,3-dipalmitate (GD) purified from Lactobacillus paracasei subsp. paracasei NTU 101-fermented products has been demonstrated to possess neuroprotective properties. We determined the effect of GD on oxygen-glucose deprivation and reperfusion (OGD/R)-induced SH-SY5Y neuroblastoma cell death. GD ameliorated OGD/R-induced apoptosis by elevating the protein expression of nuclear peroxisome proliferator-activated receptor γ (PPARγ) and nuclear factor erythroid 2-related factor 2 (Nrf2), thereby attenuating reactive oxygen species (ROS) generation. Pretreatment with GD reduced nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) expression from 1.54 ± 0.27 to 0.84 ± 0.46, thereby attenuating the induction of pro-inflammatory mediators, and increased the plasma membrane Ca2+ ATPase (PMCA) levels from 0.81 ± 0.02 to 1.08 ± 0.06, thus reducing the levels of cytosolic Ca2+; this also correlated with reduced cell death. We conclude that GD prevents SH-SY5Y cells from injury after OGD/R insult, possibly by modulating oxidative stress and inflammatory response.

Prevention of hypertension-induced vascular dementia by Lactobacillus paracasei subsp. paracasei NTU 101-fermented products.

CONTEXT: Numerous etiological studies have established positive clinical association between hypertension and vascular dementia (VaD). Lactobacillus paracasei subsp. paracasei NTU 101-fermented products have been shown to decrease vascular risk factors such as hypertension, atherosclerosis, hyperlipidemia and obesity. OBJECTIVE: This study investigated the effect of ethanol extract of Lactobacillus paracasei subsp. paracasei NTU 101-fermented products (NTU101F) in hypertension-induced VaD in rats. MATERIALS AND METHODS: Hypertension was promoted by subcutaneous injection of deoxycorticosterone acetate (DOCA, 25 mg/kg body weight/day, twice a week) and substitution of drinking water with 1.0% NaCl and 0.2% KCl. The NTU101F groups (0.5, 1.0, and 5.0) administered NTU101F at the concentrations 11, 22, and 110 mg/kg body weight/day, respectively, starting from day 51 day of DOCA-salt treatment. Morris water maze (MWM) was used for testing learning and memory. Different biochemical estimations were used to assess oxidative stress and inflammatory response in hippocampus. RESULTS: Oral administration of NTU101F in DOCA-salt hypertension-induced VaD rats resulted in a significant decrease in blood pressure by 18.3-23.2% (p < 0.001), which was regulated by increasing eNOS density (about 3-fold) in the aorta, promoting NO production, and decreasing of matrix metallopeptidase 9 activity (about 2-fold) in the hippocampus, in addition to improve the kidney function and structure, decrease escape latency and increase the times spent in the target quadrant by 23.5-27.8% (p < 0.05). CONCLUSION: Overall, our findings suggest that NTU101F could exert neuroprotection in the brain and attenuate hypertension-induced VaD.

Anti-obesity activity of the water extract of Lactobacillus paracasei subsp. paracasei NTU 101 fermented soy milk products.

The anti-obesity activity of the water extract of soy milk fermented with Lactobacillus paracasei subsp. paracasei NTU 101 (W101) was investigated. A high-fat diet (HFD) was used to induce obesity in rats, and the effects of daily W101 feeding (8 weeks) were observed. The rats fed the HFD and supplemented with low-dose W101 (LW101, 15 mg per kg body weight per day) or high-dose W101 (HW101, 150 mg per kg body weight per day) had significantly reduced final body weight in comparison with that of the HFD group. W101 decreased the formation of lipid plaques in the aorta, reduced the adipocyte cross-sectional area and diameter, and reduced the levels of CCAAT/enhancer-binding protein ß (C/EBPß), peroxisome proliferator associated receptor γ (PPARγ), and C/EBPα. Regarding lipogenesis regulation in adipocytes, W101 suppressed heparin-releasable lipoprotein lipase (HR-LPL) in adipose tissues and inhibited lipid absorption, thereby reducing lipogenesis. Lactobacillus paracasei subsp. paracasei NTU 101-fermented soy milk may be used to develop health foods that prevent obesity.

A cell-based model of bone remodeling for identifying activity of icarrin in the treatment of osteoporosis.

The activity of icarrin (a flavonoid from Herba epimedii) was investigated in the regulation of bone remodeling, a process coupled by osteoblast-mediated bone forming and osteoclast-mediated bone resorption. By directly co-culturing mouse bone marrow stromal cells and mouse preosteoclastic RAW264.7, and transwell co-culturing rat ovarian follicular granulosa cells (FGC), a 30 % increase in alkaline phosphatase (ALP) activity and 25 % increase in estradiol level occurred. Compared with the antiresorptive drug, alendronate, and an anabolic drug, PTH1-34, icarrin possessed all of the positive effects on the co-culture by increasing ALP activity, estradiol production and decreasing tartrate-resistant acid phosphatase activity. A similar action of icarrin occurred on co-culture of mesenchymal stem cells, mouse peripheral blood mononuclear cells, and FGC. Overall, by using a co-cultured cell-based in vitro screening assay, icarrin is suggested as a new class of dual-action therapeutic agent for osteoporosis.

Synergistic inhibitory effect of Icariside II with Icaritin from Herba Epimedii on pre-osteoclastic RAW264.7 cell growth.

Increasing evidence shows the therapeutic superiority of herbal extracts in comparison to isolated single constituents. One of the reasons may be attributed to the synergy effect of compound combinations. Flavonoids from Herba Epimedii have been shown to have therapeutic effect against bone loss. Our previous study showed that Icariside II inhibited pre-osteoclast RAW264.7 growth. The aim of this study was to investigate whether the activity of Icariside II is synergized by other components of Herba Epimedii. The inhibitory activity of Icariside II was significantly enhanced in the presence of the extract of Herba Epimedii (EHE) at the ratio of 1:1, 1:5 and 1:10. Icaritin, another flavonoid constituent, was shown here to inhibit RAW264.7 growth in a dose-dependent manner. Further, we found that Icariside II, together with Icaritin, synergistically inhibited RAW264.7 growth. The synergistic effect is significant when the ratio of Icariside II and Icaritin was 10:1, 5:1, 1:1, 1:2, and 1:5, respectively. In conclusion, Icaritin were an active component. The inhibitory activity of Icariside II on pre-osteoclast RAW264.7 growth was synergized by Icaritin, which maybe contribute to the efficiency of Herba Epimedii extract on curing bone-related diseases, such as osteoporosis.

Cytotoxic biflavones from Stellera chamaejasme.

Bioassay-guided phytochemical studies on Stellera chamaejasme led to the isolation of two new biflavones, chamaejasmenin E (1) and chamaejasmin D (2), together with ten known compounds. The structures of new compounds were elucidated by extensive spectroscopic analyses and their absolute configurations on 2, 3, 2″ and 3″ were confirmed by TDDFT quantum chemical calculated ECD spectra combined with experimental ECD spectra. All isolated biflavones were evaluated for their cytotoxic activities against Bel-7402 and A549 tumor cell lines, and sikokianin D (3) was found to possess the most potential cytotoxic activities against both the two cell lines with IC50 values of 1.29 ± 0.21 and 0.75 ± 0.25 µM, respectively. Moreover, some structure-function relationships of these bioflavones for cytotoxic activities were explored and summarized.

Inhibitory effect of Ecliptae herba extract and its component wedelolactone on pre-osteoclastic proliferation and differentiation.

ETHNOPHARMACOLOGICAL RELEVANCE: Ecliptae herba, also known as "Mo-Han-Lian", has long been used in China to nourish Kidney and thereafter strengthen bones. Accumulating evidence indicates that extracts of Ecliptae herba have antiosteoporotic effect. However, the effective compounds and cellular mode of action are still unclear. To investigate the effect of ethyl acetate extract of Ecliptae herba (EAE) and its component wedelolactone on proliferation and differentiation of preosteoclastic RAW264.7 cells as well as proliferation of bone marrow stromal cells (BMSC). MATERIALS AND METHODS: RAW264.7 and BMSC were examined for proliferation by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method. Tartrate-resistant acid phosphatase (TRAP) activity of RAW264.7 was measured by using p-nitrophenyl sodium phosphate (pNPP) assay after the cells were treated with 30ng/ml receptor activator for nuclear factor-κ B ligand (RANKL) plus various concentrations of EAE, wedelolactone or alendronate. The formation of multinucleated TRAP-positive RAW264.7 cells was observed by using a TRAP-staining kit. RESULTS: Treatment of RAW264.7 cells with EAE at high doses (20µg/ml and 40µg/ml) or wedelolactone at 10µg/ml resulted in a decrease in proliferation of RAW264.7 cells. Low doses of EAE (5, 10µg/ml) and wedelolactone (2.5µg/ml) inhibited RANKL-induced TRAP activity by 20.3%, 37.9%, and 48.3%. The inhibitory effect of wedelolactone is more potent than that of alendronate, an anti-resorptive drug. Morphological changes revealed that 5µg/ml EAE and 2.5µg/ml wedelolactone reduced the number of multinucleated osteoclast-like cells. At the high doses, EAE (20µg/ml) and wedelolactone (10µg/ml) inhibited the growth of BMSC. CONCLUSIONS: EAE and its component wedelolactone inhibited osteoclast RAW264.7 proliferation and differentiation at the low doses, but at the high doses, showed cytotoxic effect on BMSC. These results indicated that EAE and wedelolatone might be potential alternative therapy for osteoporosis.

[Antitumor components screening of Stellera chamaejasme L. under the case of discrete distribution of active data].

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.

Structure elucidation of degradation products of Z-ligustilide by UPLC-QTOF-MS and NMR spectroscopy.

Z-Ligustilide, a major phthalide isolated from a widely used traditional Chinese medicine Ligusticum chuanxiong, possesses various pharmacological activities including neuroprotective, anti-inflammatory, antiproliferative and vasorelaxing effects. However, it is unstable and inclined to degrade in natural conditions, which limits its study and application greatly. In this study, degradation behavior of Z-ligustilide and its degradation products stored at room temperature under direct sunlight were investigated and structure elucidated by HPLC-UV, UPLC-QTOF-MS and NMR. Z-ligustilide degradation and total five degradation products were generated and detected. Two degradation products were unequivocally identified as senkyunolide I and senkyunolide H by comparison with reference compounds. Another two degradation products were further isolated by semi-preparative HPLC and structure elucidated as (E)-6, 7-trans-dihydroxyligustilide and (Z)-6, 7-epoxyligustilide by 1H and 13C NMR, respectively. The degradation pathways of Z-ligustilide were finally proposed. Oxidation, hydrolysis and isomerization are the major degradation reactions.

[Analysis of chemical constituents of chuanxiong rhizoma absorbed into rat brain tissues by UPLC-Q-TOF-MS].

OBJECTIVE: To analyze and identify the chemical constituents in rat brain tissues after oral administration of Chuanxiong Rhizoma extracts. METHOD: The dosed and blank rat brain tissues were analyzed by UPLC-Q-TOF-MS. Different peaks were observed in total ion chromatograms and then identified according to their retention time, accurate mass weight, MS and MS/MS data. RESULT: After oral administration of Chuanxiong Rhizoma extracts, 3 compounds were absorbed into rat brain tissues through BBB. They were identified as senkyunolide I, senkyunolide A and ligustilide. CONCLUSION: The study is helpful for interpreting effective substance of Ligusticum chuanxiong.

A new diterpenoid from Rhizophora apiculata.

A new diterpenoid, 15(S)-isopimar-7-en-1-oxo-15,16-diol (1), was isolated from the stems of mangrove plant Rhizophora apiculata. The structure of the new compound was elucidated by MS, IR, 1D, and 2D NMR techniques, including HMQC, HMBC, and NOESY correlations. In addition, seven known constituents were isolated from this plant for the first time.

Rhizoma coptidis and berberine-induced activation of murine microglia N9 cells.

AIMS OF THE STUDY: To investigate the effect of water extract of Rhizoma coptidis (WEC) and berberine on the activation of murine microglia N9 cells and corresponding mechanism related to mitochondria. MATERIALS AND METHODS: Phagocytic activity of murine microglia N9 cells was measured by neutral red staining method after the cells were treated with various concentrations of WEC and alkaloids for 24h. Flow cytometric analysis was performed to determine the level of intracellular ROS, Ca(2+), and mitochondrial transmembrane potential (Delta psi) after 87 microg/ml of WEC and 12.4 microg/ml of berberine treatment. Global changes of gene expression in WEC- and berberine-treated N9 cells were measured using cDNA microarray. RESULTS: WEC and berberine, but not palmatine and jatrorrhizine, enhanced phagocytic activity of murine N9 cells in a dose-dependent manner. Both of WEC and berberine stimulated free radical generation, enhanced mitochondrial Delta psi and induced gene expression of Ndufab1, Cox6a2 and Atp5a1. However, a more significant phagocytic effect was observed for WEC. WEC, but not berberine, increased intracellular Ca(2+) concentration. The gene expression of Atp5c1 was selectively up-regulated by WEC, while three genes of Uqcrq, Cox8b, and Atp5g2 were induced by berberine. CONCLUSIONS: WEC and berberine activated murine microglia N9 cells by the regulation of mitochondrial function and mitochondria-related signal molecules. The action of WEC is stronger than that of berberine, indicating that the effect of WEC is ascribed partially, but not totally, to berberine.

Functional analysis of cultured neural cells for evaluating cold/cool- and hot/warm-natured Chinese herbs.

Recently, modern scientific research has been required to understand pharmacological basis of traditional Chinese medicine (TCM) theory based on the ancient clinical experience, and to investigate the molecular mechanisms of action of Chinese herbs. Here, 20 Chinese herbs, classified into 4 properties (hot, warm, cold and cool) of TCM, were analyzed for their ability to exhibit antioxidant action, to enhance glucose uptake by murine microglia N9 cells, and to influence neurotransmitter norepinephrine (NE) release from rat pheochromocytoma PC12 cells. We found a generally protective effect of both hot/warm-natured and cold/cool-natured herbs against H(2)O(2)-induced N9 cell death, partially by elevating superoxide dismutase (SOD) activity. Glucose uptake was elevated after treatment with some hot/warm-natured herbs. In addition, most herbs with hot/warm nature tended to stimulate NE release, while such stimulatory effect was not observed in the herbs with cold/cool nature. Two cold/cool-natured herbs, Rhizoma coptidis and Radix scutellariae, even significantly suppressed the release. These results suggest that the distinct abilities of Chinese herbs to regulate neural cell functions appear to be correlated with their natures identified in traditional TCM theory, and may be a useful guide for their utility in neural degenerative diseases.