Mechanism of Fat Emulsion-mediated PTEN-PI3K-AKT Signaling Pathway on Neurotoxicity of Bupivacaine.

This study was to explore the application value of chromosome ten (PTEN) - phosphatidylinositol 3-kinase (PI3K) - protein kinase B (AKT) signaling pathway in the treatment of Bupivacaine toxicity to neuronal cells under the regulation of fat emulsion. Neurons in the hippocampus of newborn rats were treated with Bupivacaine and fat emulsion and divided into five groups. The activity and action potential of neurons in each group were measured and Nissl's staining was performed. The results showed that the neuron activity of Bupivacaine group (42.36 ± 5.48%), Bupivacaine + fat emulsion group (70.23 ± 3.66%), and Bupivacaine + fat emulsion + PTEN/PI3K/AKT inhibitor group (79.28 ± 5.14%) was lower than that of the blank group (99.95 ± 3.42%). The duration of action potential in Bupivacaine group was increased (5.19 ± 0.48ms) and the frequency of action potential was decreased (13.87 ± 1.95) compared with the blank group (2.44 ± 0.37ms, 19.59 ± 2.14). The duration of the fat emulsion group (2.39 ± 0.39ms, 19.76 ± 2.05), Bupivacaine + fat emulsion group (2.88 ± 0.52ms, 18.53 ± 1.66), and Bupivacaine + fat emulsion + PTEN/PI3K/AKT inhibitor group (3.43 ± 0.69ms, 17.57 ± 1.58) was decreased, but the number of times increased (P < 0.05). In short, the fat emulsion can reverse the toxic effects of Bupivacaine on rat hippocampal neurons by regulating the PTEN/PI3K/AKT signaling pathway. This study provided a reference for the clinical treatment of the neurotoxicity of Bupivacaine.

Neuronal activity-induced SUMOylation of Akt1 by PIAS3 is required for long-term potentiation of synaptic transmission.

Neuronal activity regulates spatial distribution of the SUMOylation system in cytosolic and dendritic sites, which has been implicated in learning, memory, and underlying synaptic structural and functional remodeling in the hippocampus. However, the functional target proteins for activated small ubiquitin-like modifiers (SUMOs) and downstream molecular consequences behind long-term potentiation (LTP) of synaptic plasticity remain to be elucidated. In this study, we showed that N-methyl-D-aspartate receptor-mediated neuronal activity induced the covalent modification of cytosolic Akt1 by small ubiquitin-like modifier 1 (SUMO1) in rat cortical and hippocampal CA1 neurons. Protein inhibitor of activated STAT3 (PIAS3) was involved in the activity-induced Akt1 SUMO1-ylation, and K64 and K276 residues were major SUMOylated sites. Importantly, Akt1 SUMOylation at K64 and K276 enhanced its enzymatic activity and facilitated T308 phosphorylation. Furthermore, the N-terminal SAP domain of PIAS3 bound Akt1 directly. The disruption of Akt1-PIAS3 interaction by Tat-SAP, a synthetic Tat-fused cell-permeable peptide containing PIAS3 SAP domain, inhibited neuronal activity-induced Akt1 SUMOylation and impaired LTP expression and late phase LTP maintenance in the hippocampus. Correlatedly, Tat-SAP not only blocked the LTP-related extracellular signal-regulated kinase (ERK)1/2-Elk-1-brain-derived neurotrophic factor (BDNF)/Arc signaling, but also disrupted mammalian target of rapamycin (mTOR)-eIF4E-binding protein 1 (4E-BP1) pathway. These findings reveal an activity-induced Akt1 SUMOylation by PIAS3 that contributes to ERK1/2-BDNF/Arc and mTOR-4E-BP1 cascades, and in turn, long-lasting excitatory synaptic responses.

Long ncRNA MALAT1 promotes cell proliferation, migration, and invasion in prostate cancer via sponging miR-145.

BACKGROUND: The long non-coding (lncRNA) RNA MALAT1 (metastasis-associated lung adenocarcinoma transcript 1) is known to promote tumorigenesis, whereas microRNA-145 (miR-145) plays an antitumor role in several cancers. In this study, we aimed to elucidate the role of MALAT1 and miR-145 in prostate cancer cells and investigate the effect of MALAT1 downregulation on prostate cancer (PCa) cells in vitro in vivo. METHODS: The Cancer Genome Atlas (TCGA) datasets were used to carry out the initial bioinformatics analysis; the findings were then tested in LNCaP and CWR22Rv1 cell lines. Western blot and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to evaluate the levels of MALAT1 and miR-145 along with related biomarkers. Furthermore, wound-healing and Transwell assays were performed to test the migratory and invasive abilities of PCa cells. Luciferase reporter assays were used to validate the relationship between MALAT1 and miR-145; their down-stream target genes were also studied. To further substantiate these findings in an animal model, tumor studies including immunofluorescence staining of tissues were carried in nude mice. RESULTS: The expression of MALAT1 was upregulated in both LNCaP cell lines and CWR22Rv1 cell lines (F=2.882, t=13.370, P<0.001; F=2.268, t=15.859, P<0.001). Knockdown of MALAT1 reduced the migratory and invasive capabilities of PCa cells (F=0.017, t=12.212, P<0.001; F=10.723, t=6.016, P=0.002). Using direct binding, MALAT1 suppressed the antitumor function of miR-145, which in turn upregulated transforming growth factor-ß1 (TGF-ß1)-induced epithelial-mesenchymal transition (EMT) via SMAD3 and TGFBR2 (F=2.097, t=5.389, P=0.006; F=1.306, t=4.155, P=0.014). CONCLUSIONS: We confirmed that MALAT1 acts as a competing endogenous RNA (ceRNA) of miR-145. The MALAT1 based regulation of MiR-145-5p-SMAD3/TGFBR2 interactions could be an intriguing molecular pathway for the progression of PCa.

Intercellular adhesion molecule-1 and S-100ß in sevoflurane combined with epidural anesthesia for radical resection of lung cancer.

Significance of intercellular adhesion molecule-1 (ICAM-1) and S-100ß protein (S-100ß) were investigated in sevoflurane combined with epidural anesthesia for radical resection of lung cancer. In total, 172 patients who underwent radical resection of lung cancer from May 2014 to January 2016 and 160 blood samples from healthy patients in the same period were selected for prospective analysis. Lung cancer patients were the therapy group (TG). Venous blood (2 ml) was taken from patients before anesthesia (T1) and after anesthesia at 30 min (T2), 3 h (T3) and 24 h (T4), respectively. Healthy physical examination samples were the control group (CG). Enzyme linked immunosorbent assay (ELISA) was used to detect the concentration of ICAM-1 and RT-PCR to detect the concentration of S-100ß. The changes of ICAM-1 and S-100ß concentrations and their significance to patients were analyzed. The serum ICAM-1 and S-100ß concentrations in TG were significantly higher than those in CG (P<0.001), and were the highest at T1 (P>0.001), followed by T2 (P<0.001). Of the 172 patients 27 cases had adverse complications during the perioperative period. Patients were divided into the ICAM-1 high concentration group (CHCG), the ICAM-1 low concentration group (CLCG) and the S-100ß high concentration group (SHCG) and the S-100ß low concentration group (SLCG). The 3-year survival rate of CHCG was significantly lower than CLCG and SLCG (P<0.001). ICAM-1 and S-100ß protein in sevoflurane combined with epidural anesthesia for radical resection of lung cancer can effectively predict the perioperative adverse complications of patients, and have better monitoring significance for the prognosis of patients.

Trolline Ameliorates Liver Fibrosis by Inhibiting the NF-κB Pathway, Promoting HSC Apoptosis and Suppressing Autophagy.

BACKGROUND/AIMS: Previous studies have shown that trolline possesses various forms of pharmacological activity, including antibacterial and antiviral potency. The present paper addressed the putative hepatoprotective effects of trolline. METHODS: Rats received 2 ml/kg CCl4 (mixed 1: 1 in peanut oil) intragastrically twice a week for 8 weeks to induce hepatic fibrosis. The animals were then treated with trolline for additional 4 weeks. Liver pathology and collagen accumulation were observed by hematoxylin-eosin and Masson's trichrome staining, respectively. Serum transaminase activity and collagen-related indicator level were determined by commercially available kits. NF-κB pathway activation was also examined. Moreover, the effects of trolline on hepatic stellate cell (HSC-T6) apoptosis, mitochondrial membrane potential (MMP), and autophagy were assessed. RESULTS: Trolline significantly alleviated CCl4-induced liver injury and notably reduced the accumulation of collagen in liver tissues. Trolline treatment also markedly decreased inflammatory cytokines levels by inhibiting the NF-κB pathway. Trolline strongly inhibited HSC-T6 activation and notably induced cell apoptosis by modulating the Bax/Bcl-2 ratio, caspase activity, and MMP. Moreover, trolline significantly inhibited HSC-T6 autophagy, as evidenced by the decrease in the formation of autophagic vacuoles and the number of autophagosomes, by regulating the expression levles of LC3, Beclin-1, P62, Atg 5 and 7. CONCLUSION: Our study demonstrates that trolline ameliorates liver fibrosis, possibly by inhibiting the NF-κB pathway, promoting HSCs apoptosis and suppressing autophagy.

Raf Kinase Inhibitory Protein Down-Expression Exacerbates Hepatic Fibrosis In Vivo and In Vitro.

BACKGROUND/AIMS: Raf kinase inhibitory protein (RKIP) is closely associated with numerous tumors and participates in their development through regulating the growth, apoptosis, invasion and metastasis of tumor cells. However, the role of RKIP in chronic liver injury and particularly in liver fibrosis is still unclear. METHODS: In the present study, hepatic fibrosis was induced by porcine serum (PS) in rats and primary hepatic stellate cells (HSCs) were isolated from rat livers. Moreover, locostatin was used to interfere with RKIP expression. RESULTS: RKIP expression was significantly inhibited by locostatin in both liver tissues of rats and primary HSCs. Down-regulating RKIP expression resulted in serious liver injury, extensive accumulation of collagen, and significant increase in the levels of ALT, AST and TNF-α during liver fibrosis in rats. Moreover, down-regulating RKIP significantly promoted HSCs proliferation and colony formation in vitro. Reduced RKIP significantly increased the production of collagen and the level of α-SMA as well as the expression of MMP-1 and MMP-2 in both liver tissues and primary HSCs. Furthermore, down-regulating RKIP promoted the activation of the ERK and TLR4 signaling pathways. CONCLUSION: Our findings clearly indicate an inverse correlation between RKIP level and the degree of the liver injury and fibrosis. The decrease in RKIP expression may exacerbate chronic liver injury and liver fibrosis.

Loss of Raf kinase inhibitor protein is associated with malignant progression in hepatic fibrosis.

Raf kinase inhibitory protein (RKIP), besides regulating important intracellular signaling cascades, was described to be associated with progression, metastasis and prognosis in several human neoplasms. But its role in hepatic fibrogenesis remains unclear. In the present study, we found that the absence of RKIP expression significantly enhanced the proliferation of HSC-T6 cells. Reduced RKIP expression promoted the activation of HSCs and the accumulation of collagen, as evidenced by the increases in the levels of collagen I and α-smooth muscle actin. Moreover, down-regulating RKIP expression led to severe histopathological changes and collagen accumulation in hepatic tissues of rats with liver fibrosis. Furthermore, the absence of RKIP promoted the activation of ERK/MAPK pathway in vitro and in vivo. Our findings clearly demonstrate an inverse correlation between RKIP level and the degree of the liver injury and fibrosis. Loss of RKIP may be associated with malignant progression in hepatic fibrosis.