Folia histochemica et cytobiologica最新文献

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: 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.

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: Standard treatment for chronic lymphocytic leukemia (CLL) has experienced a dramatic change over the last few years. Until recently, CLL was treated using chemoimmunotherapy (CIT) with anti-CD20 monoclonal antibodies. Even though novel agents such as BTKi (Bruton Tyrosine Kinase inhibitor) and BCL2 inhibitors are the standard of care in most therapeutic settings, CIT still has its place in CLL treatment. Interestingly, little is known about its effects on the immune system of patients with CLL. Contrary to the reduction of the number of CLL cells during CIT administration, little attention has been paid to the cellular microenvironment, the evaluation of which during treatment may provide additional information about the course of the disease and prognosis and therefore was set as the aim of this study.

Material and methods: Flow cytometry was used to evaluate the phenotypes of different populations and subpopulations of lymphocytes in the peripheral blood (PB) of 20 patients with CLL before, during, and after CIT.

Results: During the CIT with R-FC (Rituximab, Fludarabine, and Cyclophosphamide) and R-B (Rituximab, Bendamustine) regimens, the sizes of the assessed populations and subpopulations of lymphocytes were dramatically reduced. Twenty-eight days after the first course of treatment, the exponential decrease of CLL cells was observed, and their number had declined to the median level of 10% of the numbers observed before the treatment. T cells, NK cells, NKCD56dim, NKT-like, and NKT-like CD56dim also decreased exponentially. After the second treatment course, a decline in the numbers of T, NK, NKCD56dim, NKT-like, and NKT-like CD56dim cells was observed, which were stable until the sixth treatment course. However, the number of NKT-like CD56bright cells decreased to the third course of treatment and then increased. The number of CLL cells in peripheral blood correlated with the number of NKCD56bright cells, influencing the treatment response.

Conclusions: Upon CIT, the reduction of CLL cells is accompanied by shifts in immune cell populations, T, NK, and NKT-like cells. Monitoring changes of those cell populations in the peripheral blood may serve as an important predictive and prognostic indicator.

Introduction: This study is to detect the expression of inflammatory factor or neutrophil-activating factor IL-8 and Wnt2 in gastric cancer (GC) and investigate the involvement of IL-8 and Wnt2 expressions in the clinicopathological indexes and prognosis.

Material and methods: We detected the expression of IL-8 and Wnt2 in 100 GC tissues and 40 normal gastric mucosae using immunohistochemistry. The relationships between the IL-8 and Wnt2 expression and the clinicopathological characteristics were explored. The relationship between IL-8 expression, Wnt2 expression, and prognosis of GC was analyzed by survival curve and survival regression.

Results: The expression of IL-8 and Wnt2 in GC tissue was 64% and 75% respectively, which was significantly higher than that in adjacent normal gastric mucosa tissues, moreover, expressions of IL-8 and Wnt2 were positively correlated. The positive rate of IL-8 and Wnt2 expressions were correlated with lymph node metastasis and TNM staging （P < 0.01, and Wnt2 was also correlated with infiltration depth (P = 0.021), but there was no difference with age, sex, and differentiation (P > 0.05). The 3-year survival analysis showed that the survival rates of IL-8- and Wnt2-positive patients were 20% and 24%, respectively, which were significantly lower than those of negative patients. Cox regression analysis showed that IL-8 and Wnt2 may be independent factors affecting the prognosis of GC.

Conclusions: Our data demonstrated that the overexpression of IL-8 and Wnt2 could be isolated prognostic factors in patients with GC and, possibly, may present new targets for the treatment of GC.

Introduction: Pre-eclampsia is a pregnancy-specific syndrome, which is partly due to abnormal proliferation and invasion of trophoblast cells. EP300 interacting inhibitor of differentiation 1 (EID1) participates in cell proliferation and invasion. This study aims to investigate the roles of EID1 in trophoblast cells and pre-eclampsia.

Material and methods: The expression of EID1 in placental tissues from 60 women with pre-eclampsia and 60 health pregnancies was detected by real-time PCR and immunohistochemical staining. EID1 was overexpressed or silenced by transfection of plasmid or siRNA in HTR-8/SVneo trophoblast cells, and then cell proliferation, cell cycle transition, migration, and invasion were determined by CCK-8 assay, flow cytometry, immunofluorescent staining, immunoblotting, and transwell assays. In addition, the activity of Akt/b-catenin signaling was measured by immunofluorescent staining and Western blot.

Results: EID1 mRNA level was decreased in placental tissues of pre-eclampsia patients, especially early-onset pre-eclampsia, accompanied by more severe clinical manifestation and a higher rate of fetal growth restriction (FGR). Gain- and loss-of-function experiments demonstrated that EID1 promoted proliferation and cell cycle transition, migration, and invasion in HTR-8/SVneo cells and its knockdown played opposite roles, suggesting that EID1 may be required for normal gestation. Akt/b-catenin signaling was activated after EID1 forced expression and deactivated after its silencing.

Conclusions: EID1 promoted proliferation and invasion of cultured trophoblast cells with possible involvement of Akt/b-catenin signaling. These findings may provide novel insights for the diagnosis and treatment of early-onset pre-eclampsia in a clinic.

Introduction: Ulcerative colitis (UC) is a nonspecific intestinal inflammatory disease. Dexmedetomidine (DEX) is a selective alpha 2-adrenergic receptor agonist commonly used for analgesia and sedation in intensive care units. Herein, the role and mechanism of DEX in dextran sulfate sodium (DSS)-induced colitis was explored.

Materials and methods: A murine model of DSS-induced colitis was established by adding 3.5% (w/v) DSS in drinking water to C57BL/6J female mice. The severity of colitis was measured by the disease activity index (DAI) score, colon length and body weight of mice. The serum concentration and mRNA levels of inflammatory cytokines in colon tissues were assessed by ELISA and RT-qPCR, respectively. Protein levels of apoptotic markers, tight junction proteins and genes involved in the TLR4/MyD88/NF-κB signaling were quantified utilizing Western blotting. The pathological changes of colon tissues were evaluated by hematoxylin-eosin (HE) staining and histological score. Intestinal permeability in vivo was assessed by fluorescein isothiocyanate (FITC)-dextran (FITC-D) administration. TUNEL assay was used to determine cell apoptosis in the intestinal epithelium.

Results: DSS administration resulted in weight loss, shortening of the colon, increased DAI score, histological abnormalities, and increased serum FITC-D levels in mice, all of which were reversed by DEX injection. Moreover, DEX attenuated DSS-triggered inflammatory response, intestinal barrier injury and intestinal epithelial cell apoptosis. Mechanically, DEX inactivated the TLR4/MyD88/NF-κB signaling in the colon tissues.

Conclusions: DEX exerts beneficial effects against the intestinal barrier dysfunction, inflammatory response, and apoptosis of intestinal epithelial cells via inactivation of the TLR4/MyD88/NF-κB signaling in mice with DSS-induced colitis.