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Our previous study isolated an anti-fatigue polysaccharide (HRWP) from the Hippophae rhamnoides berry. In this study, using ion-exchange chromatography and gel filtration chromatography in turn, a water-soluble homogenous polysaccharide HRWP-A was isolated from HRWP. Structural analysis determined that HRWP-A was a polysaccharide with repeating units of (1→4)-β-d-galactopyranosyluronic residues, of which 85.16% were esterified with methyl groups. An antitumor activity assay showed that HRWP-A could significantly inhibit the Lewis lung carcinoma (LLC) growth in tumor-bearing mice. Further experiments suggested that the antitumor effect of HRWP-A might be mediated through immunostimulating activity, as it enhances the lymphocyte proliferation, augments the macrophage activities, as well as promoting NK cell activity and CTL cytotoxicity in tumor-bearing mice. To our knowledge, this is the first report on a natural antitumor high-methoxyl homogalacturonan pectin from the H. rhamnoides berry-a compound that acts as a potential immunostimulant and anticancer adjuvant.

Ethnopharmacological relevance: The fruits of <b>Hippophae rhamnoides</b> L., <b>Lycium barbarum</b> L., <b>Lycium ruthenicum</b> Murr. and <b>Nitraria tangutorum</b> Bobr. are traditional medicinal food of Tibetans and used to alleviate fatigue caused by oxygen deficiency for thousands of years. The present study focused on exploiting natural polysaccharides with remarkable anti-fatigue activity from the four Qinghai-Tibet plateau characteristic berries.<br>Materials and methods: The fruits of <b>Hippophae rhamnoides</b>, <b>Lycium barbarum</b>, <b>Lycium ruthenicum</b> and <b>Nitraria tangutorum</b> were collected from Haixi national municipality of Mongol and Tibetan (N 36.32°, E98.11°; altitude: 3100 m), Qinghai, China. Their polysaccharides (HRWP, LBWP, LRWP and NTWP) were isolated by hot-water extraction, and purified by DEAE-Cellulose ion-exchange chromatography. The total carbohydrate, uronic acid, protein and starch contents of polysaccharides were determined by a spectrophotometric method. The molecular weight distributions of polysaccharides were determined by gel filtration chromatography. Their monosaccharide composition analysis was performed by the method of 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivatization and RP-HPLC analysis. HRWP, LBWP, LRWP and NTWP (50, 100 and 200 mg/kg) were orally administrated to mice once daily for 15 days, respectively. Anti-fatigue activity was assessed using the forced swim test (FST), and serum biochemical parameters were determined by an autoanalyzer and commercially available kits; the body and organs were also weighted.<br>Result: LBWP, LRWP and NTWP were mainly composed of glucans and some RG-I pectins, and HRWP was mainly composed of HG-type pectin and some glucans. All the four polysaccharides decreased immobility in the FST, and the effects of LBWP and NTWP were demonstrated in lower doses compared with HRWP and LRWP. There was no significant difference in liver and heart indices between non-treated and polysaccharide-treated mice, but the spleen indices were increased in LBWP and NTWP (200 mg/kg) group. Moreover, the FST-induced reduction in glucose (Glc), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and increase in creatine phosphokinase (CK), lactic dehydrogenase (LDH), blood urea nitrogen (BUN), triglyceride (TG) and malondialdehyde (MDA) levels, all indicators of fatigue, were inhibited by HRWP, LBWP, LRWP and NTWP to a certain extent while the effects of LBWP and NTWP were much better than that of HRWP and LRWP at the same dosage.<br>Conclusion: Water-soluble polysaccharides HRWP, LBWP, LRWP and NTWP, from the fruits of four Tibetan plateau indigenous berry plants, significantly exhibited anti-fatigue activities for the first time, through triglyceride (TG) (or fat) mobilization during exercise and protecting corpuscular membrane by prevention of lipid oxidation via modifying several enzyme activities. Moreover, it is demonstrated that LBWP and NTWP are more potent than HRWP and LRWP, which were proposed to be applied in functional foods for anti-fatigue and antioxidant potential.<br><br>Display Omitted

The Nitraria tangutorum Bobr. fruit is an indigenous berry of the shrub belonging to the Zygophyllaceae family which grows at an altitude of over 3000 m in the Tibetan Plateau, and has been used as a native medicinal food for treating weakness of the spleen, stomach syndrome, dyspepsia, neurasthenia, dizziness, etc. for thousands of years. Nowadays, N. tangutorum industrial juice by-products generated from health food production can be a potential low cost source of some unique bioactive ingredients. In a prior study, we established a simultaneous microwave/ultrasonic assisted enzymatic extraction method for extracting antioxidant ingredients from the industrial by-products of N. tangutorum juice. In this study, these ingredients were selectively fractionated by cation-exchange resin chromatography to obtain an anthocyanin fraction namely NJBAE. NJBAE was found to be composed of 16 anthocyanins derived from six anthocyanidins by HPLC-ESI-MS, and has an appreciable cardioprotective effect on doxorubicin-induced injured H9c2 cardiomyocytes. The cardioprotective mechanism research showed that NJBAE could directly scavenge ROS, restrict further generation of ROS, promote the activity of key antioxidase, enhance glutathione redox cycling, then affect the apoptotic signaling changes in a positive way, and finally mediate caspase-dependent cell death pathways. Therefore, NJBAE has great potential to be used for preventing and treating cardiovascular disease in the food, pharmaceutical and other emerging industries.

Our previous study isolated a natural high-methoxyl homogalacturonan (HRWP-A) from Hippophae rhamnoides and showed antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of HRWP-A were further investigated. Results showed that HRWP-A could recover the body condition and activated macrophage in Cyclophosphamide (CTX)-induced immunosuppressed mice. Further, we investigated the possible mechanism underlying the effects of HRWP-A on mouse peritoneal macrophages. qPCR and western blot revealed that HRWP-A upregulated the expression of TLR4 mRNA in vitro. This process was accompanied by a clear increase in MyD88 expression and p-IκB-α, but these effects were largely abrogated by pretreatment with anti-TLR4 antibodies. The effects of HRWP-A on macrophage NO, IL-1β and IL-6 production were also inhibited by anti-TLR4 antibodies and were greatly influenced by the NF-κB inhibitor PDTC. Moreover, HRWP-A failed to induce the production of NO, IL-1β and IL-6 in peritoneal macrophages prepared from C3H/HeJ mice, which have a point mutation in the Tlr4 gene, suggesting the involvement of the TLR4 molecule in HRWP-A-mediated macrophage activation. These results may have important implications for our understanding of the structure-activity relationship of immunopotentiating polysaccharides from medicinal herbs.

• A simultaneous microwave/ultrasonic-assisted enzymatic extraction method was established for the first time. • Simultaneous microwave/ultrasonic-assisted enzymatic process can improve antioxidant capacity of juice by-product extract. • Simultaneous microwave/ultrasonic-assisted enzymatic process can increase the extraction efficiency of antioxidant ingredients. • <b>Nitraria tangutorun</b> Bobr. juice by-products extract exhibited excellent cell protection effect from oxidative injury.<br>By-products originating from food processing are a considerable disposal problem for the food industry. Because of the absence of specifically effective processing technology, huge quantities of by-products are often abandoned as rubbish and prone to microbial spoilage. Given this, a simultaneous microwave/ultrasonic assisted enzymatic extraction (SMU-AEE) method was established for the first time, and performed for antioxidant ingredients extraction from <b>Nitraria tangutorum</b> juice by-products (NJB) in the present study. Its experimental conditions were optimized by single factor test and response surface methodology (RSM), and gave the corresponding response values for antioxidant capacity of NJB extract (NJBE) of 219.73 ± 7.03 mg TE/g, which was 27.62%-190.23% higher than those obtained by traditional extraction methods. Chemical composition assay suggested that the increasing of antioxidant capacity of NJBE by SMU-AEE was because of the improvement of extraction efficiency of antioxidant ingredients from NJB, including phenols, flavonoids and anthocyanins. Furthermore, oxidative injury protection ability assay showed that NJBE was good at protecting cells from UVB-oxidative phototoxicity and doxorubicin-oxidative cardiotoxicity, and its protecting ability surpasses or approaches to that of grape seed extract (GSE, the positive control drug), indicating its good potential to be a natural antioxidant in food and pharmaceutical industries.