Activation of Intestinal HIF2α Ameliorates Iron-Refractory Anemia.

In clinics, hepcidin levels are elevated in various anemia-related conditions, particularly in iron-refractory anemia and in high inflammatory states that suppress iron absorption, which remains an urgent unmet medical need. To identify effective treatment options for various types of iron-refractory anemia, the potential effect of hypoxia and pharmacologically-mimetic drug FG-4592 (Roxadustat) are evaluated, a hypoxia-inducible factor (HIF)-prolyl hydroxylase (PHD) inhibitor, on mouse models of iron-refractory iron-deficiency anemia (IRIDA), anemia of inflammation and 5-fluorouracil-induced chemotherapy-related anemia. The potent protective effects of both hypoxia and FG-4592 on IRIDA as well as other 2 tested mouse cohorts are found. Mechanistically, it is demonstrated that hypoxia or FG-4592 could stabilize duodenal Hif2α, leading to the activation of Fpn transcription regardless of hepcidin levels, which in turn results in increased intestinal iron absorption and the amelioration of hepcidin-activated anemias. Moreover, duodenal Hif2α overexpression fully rescues phenotypes of Tmprss6 knockout mice, and Hif2α knockout in the gut significantly delays the recovery from 5-fluorouracil-induced anemia, which can not be rescued by FG-4592 treatment. Taken together, the findings of this study provide compelling evidence that targeting intestinal hypoxia-related pathways can serve as a potential therapeutic strategy for treating a broad spectrum of anemia, especially iron refractory anemia.

Exposure interval to ambient fine particulate matter (PM2.5) collected in Southwest China induced pulmonary damage through the Janus tyrosine protein kinase-2/signal transducer and activator of transcription-3 signaling pathway both in vivo and in vitro.

PM2.5 is a well-known air pollutant threatening public health. Studies confirmed that exposure to the particles could impair pulmonary function, cause chronic obstructive pulmonary disease, and increase the incidence of lung cancer. The characteristic of PM2.5 varies across regions. The toxic function of PM2.5 in southwest China remains to be elucidated. This study aimed to investigate lung injury and its mechanisms induced by PM2.5 collected in Chengdu. Rats were administered with PM2.5 by intratracheal instillation for 4 weeks. Biochemical, cell count, and inflammation-related parameters were measured. Lung tissues were obtained for hematoxylin and eosin and Masson's trichrome staining. The expression levels of vascular endothelial growth factor (VEGF), Janus tyrosine protein kinase-2 (JAK-2), and signal transducer and activator of transcription-3 (STAT-3) were detected by immunohistochemistry assays. Meanwhile, A549 cells were treated with the PM2.5. The cell cycle, and apoptosis were measured by flow cytometry. mRNA and protein expressions of JAK-2, STAT-3, p-STAT-3, and VEGFA were detected using qPCR and Western blot analysis respectively. Results of in vivo study showed that PM2.5 induced lung pathological injury, aggravated the accumulation of inflammatory cells, and increased the serum levels of inflammatory factors. In vitro experiments showed that PM2.5 disrupted the cell growth cycle and increased cell apoptosis through the activation of the JAK-2/STAT-3 signaling pathway. Taken together, this study provided convincing experimental evidence that PM2.5 collected in southwest China could induce pulmonary injury as manifested by inflammatory response and lung fibrosis, possibly through the modulation of the JAK-2/STAT-3 signaling pathway.

A plant-based medicinal food inhibits the growth of human gastric carcinoma by reversing epithelial-mesenchymal transition via the canonical Wnt/ß-catenin signaling pathway.

BACKGROUND: Natural products, especially those with high contents of phytochemicals, are promising alternative medicines owing to their antitumor properties and few side effects. In this study, the effects of a plant-based medicinal food (PBMF) composed of six medicinal and edible plants, namely, Coix seed, Lentinula edodes, Asparagus officinalis L., Houttuynia cordata, Dandelion, and Grifola frondosa, on gastric cancer and the underlying molecular mechanisms were investigated in vivo. METHODS: A subcutaneous xenograft model of gastric cancer was successfully established in nude mice inoculated with SGC-7901 cells. The tumor-bearing mice were separately underwent with particular diets supplemented with three doses of PBMF (43.22, 86.44, and 172.88 g/kg diet) for 30 days. Tumor volumes were recorded. Histopathological changes in and apoptosis of the xenografts were evaluated by hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining, respectively. Serum levels of TNF-α, MMP-2, and MMP-9 were detected by enzyme-linked immunosorbent assay. The mRNA expression levels of ß-catenin, GSK-3ß, E-cadherin, N-cadherin, MMP-2/9, Snail, Bax, Bcl-2, Caspase-3/9, and Cyclin D1 were evaluated via real-time quantitative polymerase chain reaction. The protein expression levels of GSK-3ß, E-cadherin, N-cadherin, and Ki-67 were determined by immunohistochemistry staining. RESULTS: PBMF treatment efficiently suppressed neoplastic growth, induced apoptosis, and aggravated necrosis in the xenografts of SGC-7901 cells. PBMF treatment significantly decreased the serum levels of MMP-2 and MMP-9 and significantly increased that of TNF-α. Furthermore, PBMF treatment notably upregulated the mRNA expression levels of GSK-3ß, E-cadherin, Bax, Caspase-3, and Caspase-9 but substantially downregulated those of ß-catenin, N-cadherin, MMP-2, MMP-9, Snail, and Cyclin D1 in tumor tissues. The Bax/Bcl-2 ratio was upregulated at the mRNA level. Moreover, PBMF treatment remarkably increased the protein expression levels of GSK-3ß and E-cadherin but notably reduced those of Ki-67 and N-cadherin in tumor tissues. CONCLUSIONS: The PBMF concocted herein exerts anti-gastric cancer activities via epithelial-mesenchymal transition reversal, apoptosis induction, and proliferation inhibition. The underlying molecular mechanisms likely rely on suppressing the Wnt/ß-catenin signaling pathway.

[Marine fish bone collagen oligopeptide combined with calcium aspartate increases bone mineral density in obariectomized rats].

OBJECTIVE: To investigate the effect of marine fish collagen oligopeptide and calcium aspartate alone or in combination on bone mineral density in ovariectomized rats. METHODS: Sixty three-month-old SPF Wistar female rats were randomly divided into 6 groups according to their body weight: sham operation group, model control group(ovariectomy), calcium aspartate group(ovariectomy), marine fish collagen oligopeptide group(ovariectomy), aspartate calcium + marine fish collagen oligopeptide group(ovariectomy) and calcium carbonate control group(ovariectomy), 10 rats in each group. The sham operation group and the model control group were given the same volume of pure water by gavage, and the other groups were intragastrically administered with calcium aspartate(116. 7 mg/kg), marine fish collagen oligopeptide(250 mg/kg), calcium aspartate(116. 7 mg/kg) + marine fish collagen oligopeptide(250 mg/kg), calcium carbonate(35. 6 mg/kg), and the test substance was continuously administered for 90 days. After 90 days, the animals were sacrificed, and the liver and kidney of the rats were taken to calculate the organ coefficient and pathological examination. The rat femurs were taken to measure bone mineral density and bone calcium content and rat serum was used to determine serum calcium, phosphorus concentration and alkaline phosphatase(ALP) activity. RESULTS: Bone mineral density and bone calcium content in the model group were significantly lower than those in the sham operation group(P<0. 05), indicating that the osteoporosis model was successfully established by oophorectomy. There was no significant difference in the organ index of each group(P>0. 05), liver/kidney HE staining microscopic examination showed no abnormal changes, indicating the safety of the test substances. The bone mineral density of the aspartate calcium + marine fish collagen oligopeptide group was significantly greater than that of the model group(P<0. 05). The bone mineral density of the aspartate calcium group and the marine fish collagen oligopeptide group was larger than that of the model group, but there was no significant difference(P>0. 05). Compared with that in the model group, the calcium content of calcium aspartate + marine fish collagen oligopeptide group was significantly higher(P<0. 05). Compared with that in the calcium aspartate group, the calcium content of calcium aspartate + marine fish collagen oligopeptide group was higher(P<0. 05), there was no significant difference in serum calcium concentration between groups(P>0. 05). Compared with that in the model group, serum phosphorus concentration in the aspartate calcium group, marine fish collagen oligopeptide group, aspartate calcium + marine fish collagen oligopeptide group was significantly higher(P<0. 05) and ALP activity was significantly reduced(P<0. 05). CONCLUSION: The combination of calcium aspartate and marine fish collagen oligopeptide has a significant effect on increasing bone mineral density, also indicating that marine fish bone collagen oligopeptide could promote absorption of calcium aspartate.

A Compound of Chinese Herbs Protects against Alcoholic Liver Fibrosis in Rats via the TGF-ß1/Smad Signaling Pathway.

Alcoholic liver fibrosis (ALF) has become a major public health concern owing to its health impacts and the lack of effective treatment strategies for the disease. In this study, we investigated the effect of a compound composed of Chinese herbs Pueraria lobata (Willd.), Salvia miltiorrhiza, Schisandra chinensis, and Silybum marianum on ALF. An ALF model was established. Rats were fed with modified Lieber-Decarli alcohol liquid diet and injected with trace CCl4 at late stage. The rats were then treated with several doses of the compound. Biochemical and fibrosis-relevant parameters were measured from the sera obtained from the rats. Liver tissues were obtained for hematoxylin and eosin and Masson's trichrome staining. Matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 were determined by immunohistochemistry assays. The mRNA and protein expression levels of transforming growth factor-ß1 (TGF-ß1), Smad2, Smad3, and Smad7 on the livers were also measured by quantitative polymerase chain reaction and Western blot. Results showed that the compound treatment alleviated pathological lesions in the liver, decreased the serum levels of hyaluronan, laminin, and hydroxyproline, and diminished the expression of hepatic tissue inhibitor of metalloproteinase-1. Compound treatment also increased hepatic matrix metalloproteinase-13 expression and inhibited the TGF-ß1/Smad signaling pathway. In conclusion, the compound has a protective effect against ALF in rats, and an underlying mechanism is involved in the TGF-ß1/Smad signaling pathway.