Folia histochemica et cytobiologica最新文献

Introduction: With cellular lipid storage varying, the balance between lipid intake and lipid degradation was a must to keep healthy and determined the level of lipid droplets. Although lipid droplets accumulation had been well demonstrated in adipocytes, gene expression profiling and gene function during adipogenesis and osteoblastogenesis remain unknown.

Material and methods: Here, this work profiled gene transcriptional landscapes of lipid droplets formation during adipogenesis from human mesenchymal stem cells (hMSCs) using RNA-Seq technique. By using RNA interference (RNAi) we investigated the function of candidate genes during adipogenesis and osteoblastogenesis using Oil Red/Alizarin Red/alkaline phosphatase (ALPL) staining and qRT-PCR (quantitative real-time PCR).

Results: Eleven differentially up-regulated genes associated with lipid droplets formation were identified at 3, 5, 7, 14, 21, and 28 days during adipogenesis. Unexpectedly, APOB per se inhibiting adipogenesis weakened osteoblastogenesis and METTL7A facilitating adipogenesis negligibly inhibited osteoblastogenesis according to the phenotypic characterization of adipocytes and osteoblasts and transcriptional condition of biomarkers through lentivirus transfection assays.

Conclusions: The establishment of the gene transcriptional profiling of lipid droplets formation would provide the molecular switches of hMSCs cell fate determination and the study targets for fat metabolic diseases.

Introduction: Mesenchymal stem cells (MSCs) are an excellent donor graft source due to their potential for self-renewal and multidirectional differentiation. However, the potential mechanisms involved in MSC homing and neural differentiation are still unclear. The purpose of this study was to explore the effects of a chemokine, SDF-1a, and Wnt3a ligand on rat MSCs' migration and b-mercaptoethanol (BME)-induced neural differentiation of MSCs.

Materials and methods: MSCs were isolated from rat bone marrow and cultured in vitro to passage 3. Scratch tests and transwell assays were used to estimate the effects of SDF-1a (25 ng/mL) and Wnt3a (10 ng/mL) on the migration of MSCs. The expression of Wnt/PCP pathway proteins RhoA, c-Jun, ATF2, and Wnt3a were assessed by Western blot. The 5 mM BME-induced neural differentiation of MSCs was determined by immunofluorescence to detect neuron- and astrocyte-specific markers such as nestin, GFAP, and Olig2.

Results: Wnt3a promoted the migration ability of MSCs and regulated the expression of RhoA, c-Jun, and ATF2 proteins. MSCs could differentiate into neural stem cells and astrocytes. Wnt3a enhanced BME induced neurogenesis in MSCs by increasing the protein expression of RhoA, c-Jun, and Wnt3a.

Conclusions: The present study demonstrated that the Wnt/PCP pathway promotes migration and neural differentiation of rat MSC.

Introduction: Dopamine (DA) is a neurotransmitter/neuromodulator found in both central and peripheral nervous systems. It plays several physiological functions in some mammalian and avian species. DA has been indicated to be associated with the neuroendocrine regulation of the reproductive cycle and maternal behaviors in the female native Thai chickens. Indeed, male birds express parental behaviors as well. To date, there are no data describing the functional aspects of the DAergic system in the male native Thai chickens. Thus, the objective of this study was to elucidate the localization of tyrosine hydroxylase (TH; a DA marker) neuronal groups in the brain of the roosters.

Material and methods: The distributions of TH immunoreactivity in the brain were detected utilizing the immunohistochemical technique.

Results: TH immunoreactivity was located throughout the brain and extensively in the diencephalon and mesencephalon. The highest density of TH-immunoreactive (-ir) neurons and fibers was found within the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML). The numbers of TH-ir neurons within the nucleus anterior medialis hypothalami (AM), nucleus paraventricularis magnocellularis (PVN), nI, and ML were then compared and revealed that the numbers of TH-ir neurons within the nI and ML were significantly higher than those of the AM and PVN.

Conclusions: These present findings suggest that the DAergic neurons within the nI and ML might play an important role in the reproductive activities of the native Thai roosters. Interestingly, the DAergic system in the nI might be involved in male reproductive activities and/or parental behaviors in this equatorial species.

Introduction: The purpose of this study is to elucidate the impact of long non-coding RNA (lncRNA) MIR4435-2HG/microRNA (miR)-125b-5p/ Semaphorins 4D (Sema4D) on colorectal cancer (CRC) cell propagation and migration.

Material and methods: Sema4D expression in 73 pairs of CRC tissues and matched adjacent normal tissues was measured by qRT-PCR and western blot and its association with pathological characteristics of CRC patients was analyzed by chi-square test. Also, the expression of MIR4435-2HG, miR-125b-5p and Sema4D in CRC cell lines was detected by qRT-PCR and Western blot. Knockdown or overexpression of MIR4435-2HG, miR-125b- 5p and Sema4D were separately performed in Caco-2 and LoVo cells, and the cell propagation, migration and invasiveness were detected by cell-counting kit 8, scratch, and transwell assays.

Results: LncRNA MIR4435-2HG and Sema4D were highly expressed, while miR-125b-5p expression was decreased in CRC tissues and cells. Knockdown of MIR4435-2HG/Sema4D or overexpression of miR-125b-5p inhibited CRC cell proliferation and aggressiveness; overexpression of MIR4435-2HG/Sema4D or knockdown of miR-125b-5p prompted the malignant behaviors of cancer cells. MIR4435-2HG and Sema4D competitively bound to miR-125b-5p.

Conclusions: LncRNA MIR4435-2HG targets miR-125b-5p to upregulate Sema4D expression, and thus regulates CRC cell propagation, migration and invasiveness.

Introduction: The present study aimed to investigate the effect of homeodomain interacting protein kinase 2 (HIPK2) on pulmonary fibrosis and the probable mechanisms.

Material and methods: We constructed a mouse model of bleomycin-induced pulmonary fibrosis and up-regulated the expression of HIPK2 in the lung by in vivo transfection. Lung tissues were collected for the detection of mesenchymal markers (α-SMA, collagen I, collagen III) and the expression of β-catenin as assessed by RT-PCR, western blot, and immunohistochemistry. Mouse lung fibroblasts (MLFs) with upregulation or downregulation of HIPK2 were successfully constructed and XAV939 was used to downregulate β-catenin expression. Then, we evaluated the activation of MLFs and the Wnt/β-catenin pathway under various conditions.

Results: The results showed that in the bleomycin-induced mouse model group, the lung alveolar structure was severely damaged, the amount of collagen fibers was increased in alveolar speta, and the expression of HIPK2 in the fibrotic area was found to be reduced. After upregulating HIPK2 in the lungs of the mouse fibrosis model we found that pulmonary fibrosis was attenuated and the expression of β-catenin and mesenchymal markers was reduced. The upregulation of HIPK2 inhibited the proliferation and migration of MLFs induced by TGF-β1, promoted apoptosis of MLFs, and reduced the expression of mesenchymal markers and β-catenin. Meanwhile, downregulation of HIPK2 promoted the proliferation and migration of MLFs, inhibited apoptosis, and promoted mesenchymal markers and β-catenin expression. XAV939 treatment of MLFs silencing HIPK2 inhibited their proliferation and activation via silencing HIPK2, promoted apoptosis, and reduced interstitial markers and β-catenin expression.

Conclusions: HIPK2 can attenuate bleomycin-induced pulmonary fibrosis by inhibiting the Wnt/β-catenin pathway in mouse lung fibroblasts.

Introduction: Peroxiredoxin 6 (Prdx6) is widely expressed in mammalian tissues. Our previous study demonstrated that Prdx6 was expressed in human epididymis, present in human seminal fluid, and in spermatozoa. The protective role of Prdx6 in maintaining the viability and DNA integrity of human spermatozoa was also detected. Here, we demonstrate the potential role and mechanism of Prdx6 in human epididymis epithelial cells (HEECs).

Material and methods: Western blotting was used to measure expression levels of key proteins in the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. The malonaldehyde (MDA) levels and antioxidant capacity in HEECs were detected with the commercial kits. Digital gene expression analysis (DGE) was used to identify gene expression patterns in control and Prdx6-interference HEECs. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to validate the DGE findings.

Results: Compared to control HEECs, the expression levels of JAK1, STAT1, phosphorylated JAK1 and STAT1 were significantly increased, while the expression level of SOCS3 was significantly decreased in Prdx6-interference HEECs. The MDA level and total antioxidant capacity in Prdx6-interference HEECs were significantly increased and decreased compared to that of control, respectively. DGE analysis identified 589 up-regulated and 314 down-regulated genes (including Prdx6) in Prdx6-interference HEECs. Thirteen significantly different pathways were identified between the two groups, with the majority of genes belonging to the CCL, CXCL, IL, and IFIT family of proteins and were related to immunity. In particular, the expression levels of IL6, IL6ST, and eighteen IFN-related genes were significantly increased in Prdx6-interference HEECs compared to control HEECs.

Conclusions: We found that reduced Prdx6 expression induced higher ROS levels in HEECs, which resulted in the activation of the IL-6 receptor and IFNγ expression to induce the JAK1/STAT1 signaling pathway.