The effects of phycocyanin on bleomycin-induced pulmonary fibrosis and the intestinal microbiota in C57BL/6 mice.

Phycocyanin (PC) is a light-harvesting protein isolated from Spirulina and has health benefits for a range of diseases including pulmonary fibrosis (PF). In this study, a bleomycin-induced pulmonary fibrosis model was used to determine whether PC attenuates PF and modulates the intestinal microbiota. The results showed that PC intervention attenuated the pulmonary fibrosis, demonstrated by hematoxylin-eosin staining (HE), Masson's trichrome staining, and lung dry-wet weight ratio, and PC significantly inhibited the production of interleukin-1 beta (IL-1ß), tumor necrosis factor-α (TNF-α), and lipopolysaccharide (LPS). Additionally, intestinal microbiota analysis revealed that PC intervention significantly increased the bacterial diversity and richness. Correlation analysis indicated that 9 families and 17 genes were significantly associated with at least 1 physiological index. And PC intervention significantly decreased the bacteria which is related to inflammation and dramatically increased the SCFAs-producing bacteria and probiotics. These data indicated that PC can decrease the pro-inflammatory cytokines and regulate the intestinal microbiota in BLM-induced PF mice.

Effects of phycocyanin in modulating the intestinal microbiota of mice.

The health-promoting effects of phycocyanin (PC) have become widely accepted over the last two decades. In this study, we investigated the effects of different doses of PC in modulating the intestinal microbiota and the intestinal barrier in mice. Six-week-old male C57BL/6 mice were treated with PC for 28 days. Fecal samples were collected before and after PC intervention, and the microbiota were analyzed by 16S rRNA high-throughput sequencing. Bacterial abundance and diversity increased after PC intervention. Saccharolytic bacteria of the families Lachnospiraceae and Ruminococcaceae, which can produce butyric acid, increased after PC treatment. The family Rikenellaceae, which contains hydrogen-producing bacteria, also increased after PC intervention. The PC treatment reduced intestinal permeability and increased the intestinal barrier function, as demonstrated by hematoxylin-eosin staining and reduced serum lipopolysaccharide levels. The modulating effects on the intestinal microbiota were more favorable in the low-dose PC group.

Preparation and characterization of novel poly (vinyl alcohol)/collagen double-network hydrogels.

Hydrogels prepared by conventional methods show poor performance in many aspects. Many double network (DN) hydrogels prepared by crosslinking agents have disadvantages such as toxicity. In this work, we prepared novel DN hydrogels with fish-originated collagen (Col) and poly (vinyl alcohol) (PVA), in which self-assembly of collagen and self-crosslinking of PVA were achieved. Infrared spectra indicated the existence of double network with chemical interactions between Col and PVA. X-ray diffraction (XRD) patterns showed that characteristic peak of freeze-thaw PVA in DN hydrogel was retained. Scanning electron microscope (SEM) images before and after degradation and swelling property tests indicated that the morphology of the hydrogel was a compact meshwork, which is consistent with the high water-retention rate. The degradation rate of the DN hydrogels was controlled by the ratio of Col and PVA. Compared with a pure collagen hydrogels, the stress of DN hydrogels was greatly enhanced from 6 to 33 kPa at a strain of 40%. This study indicates that DN hydrogels prepared by Col and PVA are an ideal biomaterial for tissue engineering.

Expression and function of HSP110 family in mouse testis after vasectomy.

HSP110 functions to protect cells, tissues, and organs from noxious conditions. Vasectomy induces apoptosis in the testis; however, little is known about the reason leading to this outcome. The aim of the present study was to evaluate the expression and function of HSP110 in mouse testis after vasectomy. Following bilateral vasectomy, we used fluorescent Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to detect apoptosis, Western blotting and immunohistochemistry to examine HSP110 expression and localization. Serum antisperm antibody (AsAb) and testosterone were measured by Enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay, respectively. Expression of endoplasmic reticulum stress (ERS) sensors and downstream signaling components was measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and the phosphorylation of eIF2α and JNK was detected by Western blotting. Vasectomy induced morphologic changes, increased apoptosis in the testis, increased serum AsAb, and decreased testosterone levels. After vasectomy, ORP150 mRNA level was increased first and then decreased, Bcl-2 was decreased, and the expression of HSPA4l, GRP78, GADD153, PERK, ATF6, IRE-1, XBP-1s, Bax, Bak, and caspases and the phosphorylation of eIF2α and JNK were increased. We present that an ER stress-mediated pathway is activated and involved in apoptosis in the testis after vasectomy. HSPA4l and ORP150 may play important roles in maintaining the normal structure and function of testis.

Protective effect of HO-1 transfection against ethanol-induced osteoblast damage.

Heme oxygenase-1 (HO-1) plays important roles in anti-oxidant, anti-inflammatory and immunoregulative activities. The aim of this study was to observe if HO-1 transfection could inhibit the damage of osteoblasts induced by ethanol. HO-1 was transfected into osteoblasts via constructed plasmid. After exposure to ethanol for 24 h, cytoactivity and apoptosis of osteoblasts were measured by MTT assay and flow cytometry, respectively. Furthermore, the oxidative stress and inflammatory factors in osteoblasts were measured. Compared to positive control group, the cytoactivity of transfected osteoblasts was significantly increased, and the apoptosis rate was significantly decreased (P<0.05). At the same time, the levels of reactive oxygen species (ROS), methane dicarboxylic aldehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1) were significantly decreased (P<0.05), and superoxide dismutase (SOD) level was increased (P<0.05) in the transfected osteoblasts as compared with positive controls. These results suggest that HO-1 plays a protective role in osteoblasts, and HO-1 transfection can effectively inhibit bone damage induced by ethanol.

[Chromomycin A(2) induces apoptosis of HepG2 cells in vitro].

OBJECTIVE: To study the effect of chromomycin A(2) in inducing apoptosis of HepG2 cells and explore the molecular mechanism. METHODS: HepG2, MCF-7, A549, and 7901 cells were exposed to chromomycin A(2) and the changes in the cell viability were detected using MTT assay. The changes in the chromatins were observed with laser scanning confocal microscope after incubation of the cells with chromomycin A(2) (60 nmol/L) for 24 h. The changes in cell morphology were examined with a phase-contrast microscope, and the apoptotic cell populations, fluorescent intensity of reactive oxygen species (ROS) and mitochondrial membrane potential were determined using flow cytometry. RESULTS: Chromomycin A(2) significantly inhibited the proliferation of the cells in a time- and dose-dependent manner, and caused changes in the cell morphology and cell apoptosis. Exposure of the cells to chromomycin A(2) resulted in chromatin condensation, ROS generation, and reduction of the mitochondrial membrane potential. CONCLUSION: Increased ROS and mitochondria damage may importantly contribute to chromomycin A(2)-induced apoptosis in HepG2 cells.

[Expression of heat shock protein 90ß and its regulation in the reproductive system of male mice].

OBJECTIVE: To study the expression and regulation of heat shock protein 90ß (HSP90ß) in the testis, epididymis and sperms of mice. METHODS: The localization and expression of HSP90ß mRNA and protein were investigated in the testis, epididymis and sperms of mice, and the regulation of HSP90ß in the male reproductive system was explored. RESULTS: HSP90ß was expressed at a higher level in the epididymis than in the testis. In the sperms of the mice, HSP90ß was localized in the acrosome area. The expression of HSP90ß in mouse epididymis decreased after castration and recovered the normal level after testosterone treatment. HSP90ß expression in the testis and epididymis also underwent changes during the postnatal development of the mice. CONCLUSION: HSP90ß may play an important role in spermiogenesis and fertilization, and its expression pattern in the epididymis after castration and during the postnatal development suggests its regulation by hormones and development.

ACC oxidase (ACO) genes in Trifolium occidentale (L.) and their relationship to ACO genes in white clover (T. repens L.) and T. pallescens (L.).

The identification and expression of two ACC oxidase (ACO) genes during leaf development in Trifolium occidentale (L.), one of the putative ancestral genomes of the allotetraploid, T. repens (L.; white clover), is described. In common with observations made in T. repens, the ACO genes displayed differential expression, with a TR-ACO2-like gene (designated TO-ACO2) confined to developing and early mature-green leaf tissue while expression of a TR-ACO3-like gene (designated TO-ACO3) is highest in leaves at the onset and during senescence. Biochemical analysis of TO-ACO2 revealed that both accumulation of the protein (determined by western analysis with a TR-ACO2 antibody) and enzyme activity matched the transcriptional activity of TO-ACO2. Western analysis also revealed that the Tr-ACO2 antibody recognised a protein of 37 kDa as a putative TP-ACO2 in T. pallescens. The 3'-UTRs of TO-ACO2 and TO-ACO3 were then compared with the 3'-UTRs of a TR-ACO2-like and TR-ACO3-like gene from T. pallescens, the other proposed ancestral genome (or closely related to the ancestor) of T. repens, with identity values of 87.8% for the ACO2-like genes and 94.8% for the ACO3-like genes. Comparison of the 3'-UTRs of TO-ACO2 with a TO-ACO2-like gene in T. repens (designated TR(O)-ACO2) and TP-ACO2 with a TP-ACO2-like gene in T. repens (designated TR(P)-ACO2) revealed identities of 100% and 96.6%, respectively, lending good support to T. occidentale as one of the ancestral genomes of T. repens. A similar comparison of the 3'-UTRs of TO-ACO3 with a TO-ACO3-like gene in T. repens (designated TR(O)-ACO3) and TP-ACO3 with a TP-ACO3-like gene in T. repens (designated TR(P)-ACO3) revealed identities of 99.5% and 97.9%, respectively, again supporting T. occidentale as one of the ancestral genomes. Further, these data confirm that both TO-ACO-like and TP-ACO-like genes are expressed in the allotetraploid T. repens.