Ganoderic acid A from Ganoderma lucidum ameliorates lipid metabolism and alters gut microbiota composition in hyperlipidemic mice fed a high-fat diet.

Ganoderic acid A (GA) is one of the most abundant triterpenoids in Ganoderma lucidum, and has been proved to possess a wide range of beneficial health effects. The aim of the current study is to investigate the amelioration effects and mechanism of GA on improving hyperlipidemia in mice fed a high-fat diet (HFD). The results showed that GA intervention significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, and ameliorated the biochemical parameters of serum and liver related to lipid metabolism in HFD-fed mice. Histological analysis also showed that the excessive accumulation of lipid droplets in the liver induced by HFD-feeding was greatly alleviated by GA intervention. In addition, GA intervention also increased the level of short chain fatty acids (SCFAs) in the intestine and promoted the excretion of bile acids (BAs) through feces. High-throughput sequencing of bacterial full-length 16S rDNA revealed that daily supplementation with GA made significant structural changes in the gut microbial population of mice fed with HFD, in particular modulating the relative abundance of some function related microbial phylotypes. The relationships between lipid metabolic parameters and gut microbial phylotypes were also revealed by correlation analysis based on a heatmap and network. The result showed that 46 key gut microbial phylotypes (OTUs) were markedly correlated with at least one lipid metabolic parameter. Moreover, UPLC-QTOF/MS-based liver metabolomics showed that 111 biomarkers (47 up-regulated metabolites and 64 down-regulated metabolites) were significantly changed after high-dose GA intervention (75 mg kg-1 day-1), compared with the HFD-fed hyperlipidemic mice. Metabolic pathway enrichment analysis of the differential hepatic metabolites demonstrated that GA intervention had significant regulatory effects on primary bile acid biosynthesis, fatty acid biosynthesis, amino sugar and nucleotide sugar metabolism, inositol phosphate metabolism, and so on. In addition, GA intervention regulated the mRNA levels of hepatic genes involved in fatty acid metabolism and bile acid homeostasis. These findings present new evidence supporting that GA from G. lucidum has the potential to alleviate lipid metabolic disorders and ameliorate the imbalance of gut microflora in a positive way.

Exosomes secreted from osteocalcin-overexpressing endothelial progenitor cells promote endothelial cell angiogenesis.

Exosome secretion is an important paracrine way of endothelial progenitor cells (EPCs) to modulate resident endothelial cells. The osteocalcin (OCN)-expressing EPCs have been found to be increased in cardiovascular disease patients and are considered to be involved in the process of coronary atherosclerosis. Since OCN has been proven to prevent endothelial dysfunction, this study aimed to evaluate the effect of exosomes derived from OCN-overexpressed EPCs on endothelial cells. Exosomes derived from EPCs (Exos) and OCN-overexpressed EPCs (OCN-Exos) were isolated and incubated with rat aorta endothelial cells (RAOECs) with or without the inhibition of OCN receptor G protein-coupled receptor family C group 6 member A (GPRC6A). The effects of exosomes on the proliferation activity of endothelial cells were evaluated by CCK-8 assay, and the migration of endothelial cells was detected by wound healing assay. A tube formation assay was used to test the influence of exosomes on the angiogenesis performance of endothelial cells. Here, we presented that OCN was packed into Exos and was able to be transferred to the RAOECs via exosome incorporation, which was increased in OCN-Exos groups. Compared with Exos, OCN-Exos had better efficiency in promoting RAOEC proliferation and migration and tube formation. The promoting effects were impeded after the inhibition of GPRC6A expression in RAOECs. These data suggest that exosomes from OCN-overexpressed EPCs have a beneficial regulating effect on endothelial cells, which involved enhanced OCN-GPRC6A signaling.

Emodin azide methyl anthraquinone derivative induced G0/ G1 arrest in HER2/neu-overexpressing MDA-MB-453 breast cancer cells.

PURPOSE: Our previous data have shown that emodin azide methyl anthraquinone derivative (AMAD) triggered mitochondrial- dependent cell apoptosis involving caspase-8-mediated Bid cleavage, and induced proteasomal degradation of HER2/neu by blocking Her2/neu binding to Hsp90. In the present study, we futher investigated the effect of this compound on the cell cycle and related molecular mechanisms in HER2/neu-overexpressing MDA-MB-453 breast cancer cells. METHODS: The cell cycle distribution was tested by flow cytometry. The expression of cell cycle-related proteins was determined by Western blot analysis; DNA agarose gel electrophoresis was used to examine the apoptosis of MDAMB- 453 cells induced by emodin AMAD. RESULTS: After MDA-MB-453 cells were treated with different concentrations of emodin AMAD for 24 hrs, cells were arrested in G0/G1 phase, and the expression of G0/G1 related proteins c/Myc, Cyclin D1, CDK4 and p-Rb changed. DNA fragmentation appeared on the agarose gel in a concentration- dependent manner. CONCLUSION: Emodin AMAD induced G0/G1 arrest in Her2/ neu-overexpressing MDA-MB-453 cancer cells. This G0/G1 arrest was associated with decreasing protein expression of c-Myc, Cyclin D1, CDK4, and p-Rb.

[The influence of zi-hua burn cream on the survival of random skin flaps in rats].

OBJECTIVE: To observe the effect of zi-hua burn cream on the survival of skin flaps in rats, and its mechanisms. METHODS: 72 Wistar rats, were randomly divided into four groups as zi-hua group(n = 18, external application of alfalfa burn cream), control group (n = 18, external application of heparin sodium cream), model group (n = 18, external application of vaseline) , negative control (n = 18, no operation). 8 cm x 2 cm random skin flaps with pedicle on the side of head were designed on the back of Wistar rats. The drug was applied on the flap surface, 2 times a day. The survival of skin flaps was observed. The change of serum superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), turner necrosis factor-alpha(TNF-alpha)and interleukin-6(IL-6)were compared at 1,2,3,7 d after operation, and histologic examination was performed. RESULTS: The survival rate of zi-hua group (73.58 - 10. 74)% was significantly higher than that of model group (33.40 - 16.05) %, showing a statistical difference (Q = 10.63, P <0.01). There was no significant difference between the zi-hua group and control group (71.65 +/- 11. 92) %. The level of serum SOD, NO in zi-hua group and control group was higher than that in model group, while the level of serum MDA, TNF-alpha and IL-6 was lower than that in model group(P <0.01). On 7 day after operation, skin flaps tissue edema,necrosis and inflammatory cell infiltration in zi-hua group and control group was less obvious than that in model group. There was significant proliferation of granuloma and fibroblast and formation of neonatal capillary in zi-hua group and control group. The vascular density in zi-hua group was obviously higher than that in the model group and control group(P <0. 01). CONCLUSIONS: Zi-hua burn cream could significantly improve the blood supply of skin flaps, increase the survival rate of skin flaps in rats. Its mechanism may be associated with the anti-free-radical-damage action, improve local microcirculation, improve the NO content, reduce the TNF-alpha and IL-6 level, reduce inflammation factor release, improve oxidative stress state, and reduce inflammation reaction.

[Study of MSCs in vitro cultured on demineralized bone matrix of mongrel].

PURPOSE: To manufacture demineralized bone matrix(DBM) of mongrel and to explore the feasibility of DBM as scaffold of bone tissue engineering. METHODS: Thigh bones of mongrel were degreased, demineralized, deproteined, freezed, dried and sterilized to form DBM. Mesenchymal stem cells(MSCs) were seeded onto the scaffold and their growth were examined by inverted phase contrast microscope and scanning electron microscope. RESULTS: DBM had a three-dimensional mesh structure.The mean pore diameter of DBM was (254.39+/-88.71)microm and the pore rate was about 70%.MSCs could adhere to the surface and inner walls of DBM, proliferated well and secreted a large amount of extracellular matrix. CONCLUSION: DBM has satisfactory biocompatibility.