Portulaca oleracea L. Extract Alleviated Type 2 Diabetes Via Modulating the Gut Microbiota and Serum Branched-Chain Amino Acid Metabolism.

SCOPE: Portulaca oleracea L. extracts (PE) show hypoglycemic function, but the precise mechanism remains obscure. This study is designed to investigate the association of the antidiabetes effect of PE with the gut microbiota modulation and BCAAs metabolism. METHODS AND RESULTS: The Orbitrap LC-MS to Orbitrap Fusion Lumos Tribrid mass spectrometer is employed to analyze the major compounds in PE. The components of the intestinal microflora in diet-induced/STZ-treated diabetic mice are analyzed by high-throughput 16S rRNA genes sequencing. The results show that PE improves blood glucose and insulin level, increases anti-inflammatory cytokine level, lowers serum branched-chain amino acids (BCAAs), and increases serum glutamine level. PE also protects the mucosal epithelium of the colon and cecum from damage. On the impact of gut microbial composition, PE reduces the Firmicutes to Bacteroidetes ratio and the abundance of the Lachnospiraceae_NK4A136_group, Blautia, Ruminiclostridium_9, Dubosiella, and increases the abundance of the Bacteroides, Akkermansia, and Mucisprillum genera. Bacterial functionality prediction indicates PE potentially inhibits bacterial BCAAs biosynthesis, and promotes the tissue-specific expression of BCAAs catabolic enzyme for reducing BCAAs supplementation. CONCLUSION: These results reveal that PE improving T2D-related biochemical abnormalities is associated not only with gut microbiota modification but also with the tissue-specific expression of BCAAs catabolic enzyme.

A directed co-assembly of herbal small molecules into carrier-free nanodrugs for enhanced synergistic antitumor efficacy.

Carrier-free nanomedicines without structural modification are attractive for the development of natural small molecules (NSMs) and biomedical applications. Moreover, the combination of NSMs is expected to obtain nanomedicines with high efficacy and low side effects due to their inherent pharmacological activities and health benefits. However, poor water solubility and low bioavailability of NSMs limit their wider biomedical and clinical applications. In this study, we revealed the co-assembly properties of pentacyclic triterpenoids and constructed a series of carrier-free nanodrugs, which are co-assembled nanoparticles (NPs) formed by the combination of two NSMs via a supramolecular assembly strategy. Experimental work and simulation studies were combined to reveal the co-assembly mechanism of non-covalent interactions between NSMs. Not only do co-assembled NPs have rapid cellular uptake ability and passive targeting tumor ability based on the EPR effect, but also their constituent units could arrest the cell cycle at different stages of tumor cells and induce apoptosis, showing synergistic anti-tumor effects (CI < 0.7). Compared with self-assembled NPs and positive control, co-assembled NPs show the strongest therapeutic effect in vivo. Importantly, the co-assembled NPs highlight the unique advantages of NSMs in terms of biosafety and health benefits, and systemic toxicity and histological examination confirm that co-assembled NPs have reliable biosafety, and no side effects and nano toxicity risks were observed.

Anti-edema and antioxidant combination therapy for ischemic stroke via glyburide-loaded betulinic acid nanoparticles.

Stroke is a deadly disease without effective pharmacotherapies, which is due to two major reasons. First, most therapeutics cannot efficiently penetrate the brain. Second, single agent pharmacotherapy may be insufficient and effective treatment of stroke requires targeting multiple complementary targets. Here, we set to develop single component, multifunctional nanoparticles (NPs) for targeted delivery of glyburide to the brain for stroke treatment. Methods: To characterize the brain penetrability, we radiolabeled glyburide, intravenously administered it to stroke- bearing mice, and determined its accumulation in the brain using positron emission tomography-computed tomography (PET/CT). To identify functional nanomaterials to improve drug delivery to the brain, we developed a chemical extraction approach and tested it for isolation of nanomaterials from E. ulmoides, a medicinal herb. To assess the therapeutic benefits, we synthesized glyburide-loaded NPs and evaluated them in stroke- bearing mice. Results: We found that glyburide has a limited ability to penetrate the ischemic brain. We identified betulinic acid (BA) capable of forming NPs, which, after intravenous administration, efficiently penetrate the brain and significantly reduce ischemia-induced infarction as an antioxidant agent. We demonstrated that BA NPs enhance delivery of glyburide, leading to therapeutic benefits significantly greater than those achieved by either glyburide or BA NPs. Conclusion: This study suggests a new direction to identify functional nanomaterials and a simple approach to achieving anti-edema and antioxidant combination therapy. The resulting glyburide- loaded BA NPs may be translated into clinical applications to improve clinical management of stroke.

Anti-Disuse Osteoporosis Activity of a Complex of Calcium-Binding Peptide from Auricularia auricula Protein Hydrolysates.

Osteoporosis is a common metabolic bone disease that is often seen in bedridden patients and astronauts. Long-term bed rest and nonweight bearing tend to induce disuse osteoporosis. Calcium supplements are commonly used to help treat disuse osteoporosis along with medications, most of which are calcium carbonate based, but they have poor absorption effects. In this study, we prepared a novel Auricularia auricula peptide-calcium complex (AP-Ca) and evaluated its protective effects on disuse osteoporosis. In vitro assays showed that AP-Ca significantly increased the contents of calcium (P < 0.05) and the activity of alkaline phosphatase (AKP; P < 0.05) of osteoblasts cultured in a two-dimensional-rotating wall vessel. Meanwhile, supplementation with AP-Ca also inhibited the production of pro-inflammatory factors induced by the loss of stress, especially TNF-α (P < 0.05). In vivo, a mouse tail suspension (TS) model was established, and the results showed that AP-Ca helped to improve bone mineral density, bone mineral content, and bone organic content in TS mice and effectively alleviated the alteration of enzymes related to bone metabolism, including AKP (P < 0.05) and serum tartrate-resistant acid phosphatase (P < 0.05), to avoid more serious bone loss induced by TS. Furthermore, we found that AP-Ca downregulated the bone resorption-associated pro-inflammatory genes interleukin-1 (IL-1), tumor necrosis factor-α, and IL-6 by 59.53 ± 3.55%, 48.01 ± 5.68%, and 40.00 ± 5.89%, respectively (P < 0.05). In conclusion, AP-Ca showed potential to suppress bone loss induced by disuse and might be considered a new alternative to reduce the risk of disuse osteoporosis. PRACTICAL APPLICATION: This peptide-calcium complex supplement exhibited protective effects on the bone loss induced by disuse, which provided a new alternative for patients and astronauts to reduce the risk of disuse osteoporosis.

Antidiabetic effects of water-soluble Korean pine nut protein on type 2 diabetic mice.

Korean pine nut protein (PNP) has a variety of biological activities, which are good for human health, but its ability to preventing diabetes has not been reported. This study evaluated the effects of water-soluble proteins of Korean pine nut obtained from a dilute alkali extract on carbohydrate metabolism of type 2 diabetic mice on a model of diabetes induced using a high fat diet combined with streptozotocin. The results showed that the hypoglycemic effect of PNP at a middle dose was the most significant, which was 38.7% lower than that of control. The extract significantly improved the oral glucose tolerance and liver indexes, increased the activity of the carbohydrate metabolism enzymes, and regulated the expression of the function of key genes for carbohydrate metabolism. It had a positive effect on both insulin resistance and glycolytic/gluconeogenesis signaling. In conclusion, PNP can regulate fasting blood glucose, improve insulin resistance, correct the glucose metabolism disorder in diabetic mice, and have a positive regulatory role. As the functional food, it has the potential to be beneficial in the treatment of type 2 diabetes mellitus as a new hypoglycemic functional food.

Single small molecule-assembled nanoparticles mediate efficient oral drug delivery.

Oral drug delivery, which requires surviving the harsh environment in the gastrointestinal (GI) tract and penetrating the intestinal epithelium, has not been achieved using simple formulation nanoparticles (NPs). Medicinal natural products (MNPs) have been widely used in traditional medicine for disease management through oral consumption. However, most pharmacologically active compounds within MNPs do not have the properties suitable for oral applications. We hypothesize that some MNPs contain natural nanomaterials that can convert those compounds into oral formulations by forming NPs. After screening 66 MNPs, we identified five classes of small molecules that form NPs, many of which are capable of efficient drug encapsulation and GI penetration. We show that one of them, dehydrotrametenolic acid (DTA), is capable of mediating oral delivery for effective disease treatment. We determine that DTA NPs assemble through hydrogen bonding and penetrate the GI tract via apical sodium-dependent bile acid transporter. Our study reveals a novel class of single component, small molecule- assembled NPs for oral drug delivery, and suggests a novel approach to modernizing MNPs through nanomaterial discovery.

Protective mechanisms of purified polyphenols from pinecones of Pinus koraiensis on spleen tissues in tumor-bearing S180 mice in vivo.

The previous study evaluated the antitumor activity and the underlying mechanism of the purified polyphenols from pinecones of Pinus koraiensis (PPP-40) using a tumor-bearing S180 mice model. This study was designed to evaluate the protective effects of PPP-40 on spleen tissues of S180 mice in vivo. Pretreatment with PPP-40 (150 mg per kg BW per D) could significantly inhibit tumor growth, enhance spleen index and prevent the decline of haematological parameters of S180 mice induced by the tumor microenvironment. Moreover, the treatment with PPP-40 was shown to significantly inhibit splenocyte apoptosis by TUNEL staining and flow cytometry, characterized by the inhibition of splenocyte cycle (G0/G1) arrest, increase in the percentages of splenic T lymphocytes (CD3+ T cells) and T cell subsets (CD3+CD4+ and CD3+CD8+ T cells), as well as the production of T cell-related cytokines (IL-2, IL-12, and TNF-α) in splenocytes exposed to the tumor microenvironment. These effects were associated with a decrease in oxidative stress, as evidenced by the changes in the SOD, GSH-Px, GSH and MDA levels of liver and spleen tissues of S180 mice. Furthermore, the protective effect of PPP-40 on spleen tissues was deeply analyzed by detecting apoptosis-related proteins using immunohistochemistry staining. The results indicated that the protective multi-mechanisms of action also were associated with the inhibition of apoptosis through down-regulation protein expressions of Bax, caspase-9, caspase-8 caspase-3, Fas and up-regulation of the expressions of Bcl-2. These results suggested that PPP-40 is a natural antitumor agent and possesses strong immunomodulatory activities by protecting the spleen tissues of tumor-bearing S180 mice.

Characterization and bioactivity of polysaccharides obtained from pine cones of Pinus koraiensis by graded ethanol precipitation.

Pinus koraiensis polysaccharides (PKP) were extracted by hot water from P. koraiensis pine cones. Five polysaccharide fractions named PKP-A, PKP-B, PKP-C, PKP-D and PKP-E were successfully separated at final ethanol concentrations of 30%, 50%, 60%, 70% and 80%, respectively. HPLC, FT-IR, GC-MS and automatic amino-acid analysis were applied to investigate their chemical characteristics. Monosaccharide component analysis indicated that the five fractions were all composed of D-ribose, L-rhamnose, L-arabinose, D-xylose, D-mannose, D-glucose and D-galactose, but their molar ratios were quite different. HPLC results revealed that the polysaccharides precipitated by higher concentrations of ethanol solution had lower molecular masses. Moreover, the antioxidant activities of the five fractions were studied on the basis of hydroxyl radical and ABTS radical scavenging tests. The five graded polysaccharide fractions exhibited good inhibitory power, and MTT tests in vitro showed the IC50 of PKP-A and PKP-E were 1,072.5 and 2,070.0 µg · mL-1, respectively. These results demonstrated that the PKP could be a potential source of natural antioxidants or dietary supplements.

Chemical composition and antioxidant activities of three polysaccharide fractions from pine cones.

The traditional method of gas chromatography-mass spectrometry for monosaccharide component analysis with pretreatment of acetylation is described with slight modifications and verified in detail in this paper. It was then successfully applied to the quantitative analysis of component monosaccharides in polysaccharides extracted from the pine cones. The results demonstrated that the three pine cone polysaccharides all consisted of ribose, rhamnose, arabinose, xylose, mannose, glucose and galactose in different molar ratios. According to the recovery experiment, the described method was proved accurate and practical for the analysis of pine cone polysaccharides, meeting the need in the field of chemical analysis of Pinus plants. Furthermore; the chemical characteristics, such as neutral sugar, uronic acids, amino acids, molecular weights, and antioxidant activities of the polysaccharides were investigated by chemical and instrumental methods. The results showed that the chemical compositions of the polysaccharides differed from each other, especially in the content of neutral sugar and uronic acid. In the antioxidant assays, the polysaccharide fractions exhibited effective scavenging activities on ABTS radical and hydroxyl radical, with their antioxidant capabilities decreasing in the order of PKP > PAP > PSP. Therefore, although the polysaccharide fractions had little effect on superoxide radical scavenging, they still have potential to be developed as natural antioxidant agents in functional foods or medicine.

[Effect of equol of a isoflavone metabolite on menopausal symptoms].

OBJECTIVE: To explore the effect of equol of a isoflavone metabolite on menopausal women and the menopausal syndrome. METHODS: 71 menopausal women living in Harbin were assessed, and the participants were at the age of 45-55 years. The concentrations of Isoflavone and equol in first-void morning urines were analyzed by HPLC. All participants completed Kupperman questionnaire tables which were used to evaluate the severity of menopausal symptoms. The association between the equol concentrations in menopausal women and the menopausal syndrome were examine by statistical analysis method. RESULTS: The negative association between the urinary equol concentrations and the menopausal index score (P < 0.05) were found, and the negative correlation only exists in EP group after grouping at the concentration of 0.5 microg/ml equol. CONCLUSION: It was suggested that it were more higher of urinary equol concentration in menopausal women, more less of the menopausal symptoms.