Deficiency of immune-responsive gene 1 exacerbates interleukin-1beta-elicited the inflammatory response of chondrocytes via enhancing the activation of NLRP3 inflammasome.

Immune-responsive gene 1 (IRG1) is a multifunctional protein that mediates inflammatory responses in numerous pathological conditions. However, whether IRG1 has a relevance with osteoarthritis remains unaddressed. The inflammatory response of chondrocytes contributes to the progression of osteoarthritis. This study focused on assessing the functional link between IRG1 and interleukin-1beta (IL-1ß)-elicited the inflammatory response of chondrocytes. The expression levels of IRG1 increased markedly in osteoarthritis cartilage compared to normal healthy cartilage. IRG1 level also increased after IL-1ß stimulation in chondrocytes. The knockdown of IRG1 exacerbated IL-1ß-elicited apoptosis and degradation of the extracellular matrix in chondrocytes. The nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) inflammasome activation evoked by IL-1ß stimulation was enhanced in IRG1-deficient chondrocytes. Importantly, restraint of the NLRP3 inflammasome was able to diminish IRG1-deficiency-amplified effects on IL-1ß-stimulated chondrocytes. Additionally, the supplement of itaconate could ameliorate IL-1ß-induced the inflammatory response of chondrocytes and reverse any IRG1-deficiency-induced effects. Altogether, our findings document a vital role for IRG1/itaconate in settling the inflammatory response of chondrocytes via effects on the NLRP3 inflammasome.

Inhibitive Effect of Luteolin on Sevoflurane-Induced Neurotoxicity through Activation of the Autophagy Pathway by HMOX1.

Luteolin is a flavone compound occurring in a variety of medicinal plants, which is reported to have neuroprotective properties. In this study, we aimed to explore the effects of luteolin in alleviating sevoflurane-induced neurotoxicity. GeneCards and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform were employed to screen luteolin, sevoflurane, and neurotoxicity-related genes. Subsequently, we isolated primary neurons from the hippocampus of 1-day-old C57BL/6J mice and tested for cytotoxicity after treatment of different concentrations of luteolin. Next, we measured the expression of apoptosis by flow cytometry and assessed inflammation-related factors, including heme oxygenase-1 expression detected by immunohistochemical staining and neuronal apoptosis. Finally, water maze, open field, and fear conditioning tests were conducted to observe the interaction between luteolin and sevoflurane in cognitive impairment of mice. Luteolin had the lowest cytotoxicity at concentrations of 30 or 60 µg/mL; we selected 30 µg/mL for drug administration experiments in vitro. Luteolin inhibited sevoflurane-induced neuronal apoptosis and inflammatory responses through the autophagic pathway and thus ameliorated sevoflurane-induced cognitive impairment in mice. Mechanistically, luteolin up-regulated heme oxygenase-1 expression, which activated the autophagy pathway in vitro. This was confirmed by subsequent histological experiments in mice and behavioral results showing rescue cognitive impairment. Our findings uncovered an inhibitory role of luteolin in sevoflurane-induced neuronal apoptosis and inflammatory response through activation of autophagy arising from up-regulation of heme oxygenase-1, thereby alleviating sevoflurane-induced cognitive impairment in mice.

Upregulated microRNA-381-5p strengthens the effect of dexmedetomidine preconditioning to protect against myocardial ischemia-reperfusion injury in mouse models by inhibiting CHI3L1.

OBJECTIVE: Over the years, roles of microRNAs (miRNAs) in development of human diseases has been broadly investigated, while the role of dexmedetomidine (DEX) regulating miR-381-5p in myocardial ischemia-reperfusion injury (MIRI) remains largely unknown. Thus, we aimed to identify the effect of DEX on MIRI via mediating miR-381-5p. METHODS: The MIRI mice models were established by the ligation of the left anterior descending (LAD) artery and treated with miR-381-5p agomir, silenced chitinase-3-like 1 protein (CHI3L1) and DEX. The cardiac function, serum inflammatory factors, pathological changes and cardiomyocyte apoptosis of the mice' myocardial tissues were measured. The targeting relationship between miR-381-5p and CHI3L1 was predicted. RESULTS: MiR-381-5p expression was decreased while CHI3L1 expression was increased in myocardial tissues of MIRI mice. DEX preconditioning could improve cardiac function and attenuate the pathological changes, cardiomyocyte apoptosis in myocardial tissues and inflammatory response in serum of MIRI mice. MiR-381-5p agomir improved the protective impact of DEX on myocardial injury in MIRI mice. Moreover, there existed a target relation between miR-381-5p and CHI3L1. CONCLUSION: Our study demonstrated that upregulated miR-381-5p strengthens the effect of DEX preconditioning to protect against MIRI in mouse models by inhibiting CHI3L1.

Long noncoding RNA LINC01189 is associated with HCV-hepatocellular carcinoma and regulates cancer cell proliferation and chemoresistance through hsa-miR-155-5p.

INTRODUCTION AND OBJECTIVES: Emerging evidence has demonstrated that long noncoding RNAs (lncRNAs) may be closely associated with Hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC). In this study, we investigated the expression and functions of a lncRNA, LINC01189, in HCV-associated HCC. PATIENTS OR MATERIALS AND METHODS: LINC01189 expression was measured in HCC tumors, HCV-infected HCC tumors and HCV-infected HCC cells. LINC01189 was overexpressed in HCV-infected HepG2 cells to measure its function on HCV-correlated cancer proliferation. In HCC cell lines of Huh7 and Hep3B, LINC01189 was upregulated to investigate its effects on cancer cell proliferation and 5-FU chemoresistance. The competing endogenous RNA (ceRNA) target of LINC01189, human microRNA-155-5p (hsa-miR-155-5p) was probed by dual-luciferase assay and qRT-PCR. Hsa-miR-155-5p was upregulated in LINC01189-overexpessed Huh7 and Hep3B cells to investigate their epigenetic correlation on HCC development regulation. RESULTS: LINC01189 is downregulated in HCV-infected HCC tumors and cell lines. LINC01189 overexpression inhibited HCC cancer cell proliferation and 5-FU chemoresistance. Hsa-miR-155-5p was confirmed to be a ceRNA target of LINC01189 in HCC. Upregulating hsa-miR-155-5p reversed the LINC01189-mediated inhibition on HCC proliferation and 5-FU chemoresistance. CONCLUSIONS: LINC01189 downregulation is associated with HCV infection in HCC, and it has tumor-suppressing effects on HCC development through hsa-miR-155-5p.

Inhibition of microRNA-375 ameliorated ketamine-induced neurotoxicity in human embryonic stem cell derived neurons.

Excessive exposure to commonly used anesthetic agents, such as ketamine, may induce permanent damage to immature human brains. In this work, we used a human embryonic stem cell (hESC)-derived neuron model to assess the expression and function of human microRNA 735 (hsa-miR-375) in regulating ketamine-induced neural cell death and neural toxicity in vitro. In the in vitro culture, hESC-derived neurons were incubated with ketamine for 72 h. After that, cell viability, reactive oxygen species activity, neural apoptosis, neurite degeneration, and hsa-miR-375 gene expression were assessed, respectively. We found ketamine induced neural death, reactive oxygen species augmentation, neural apoptosis, neurite degeneration and hsa-miR-375 upregulation, in hESC-derived neurons. In addition, we discovered that, lentivirus-mediated mR-375 downregulation protected ketamine-induced neural cell death and neural toxicity. Also, human brain derived neurotrophic factor (BDNF) was found to be directly and reversely regulated by hsa-miR-375. Moreover, BDNF downregulation was shown to functionally reverse the protective effect of miR-375 downregulation on ketamine-induced neural cell death and neural toxicity. Overall, this work provided strong evidence showing hsa-miR-375 is an active regulator in anesthesia-induced neural cell death and neural toxicity, possibly via inverse regulation on BDNF gene.

Thiol-addition reactions and their applications in thiol recognition.

Because of the biological importance of thiols, the development of probes for thiols has been an active research area in recent years. In this review, we summarize the results of recent exciting reports regarding thiol-addition reactions and their applications in thiol recognition. The examples reported can be classified into four reaction types including 1,1, 1,2, 1,3, 1,4 addition reactions, according to their addition mechanisms, based on different Michael acceptors. In all cases, the reactions are coupled to color and/or emission changes, although some examples dealing with electrochemical recognition have also been included. The use of thiol-addition reactions is a very simple and straightforward procedure for the preparation of thiol-sensing probes.

High-mobility group box 1 contributes to mechanical allodynia and spinal astrocytic activation in a mouse model of type 2 diabetes.

Chronic pain is one of the most common complications of diabetes. However, current treatments for diabetic pain are usually unrealistic because the underlying mechanisms are far from being clear. Immerging studies have implicated immune factors as key players in the diabetic pain. High-mobility group box 1 (HMGB1) is an important mediator of inflammatory response, but its role in diabetic pain is unclear. In the present study, we observed that db/db mice (a model of type 2 diabetes) developed persistent mechanical allodynia from postnatal 2 months. Western blot showed that in postnatal 2-5 months, HMGB1 was significantly higher than that of the heterozygous littermates (db/+) mice. Intrathecal injection of a HMGB1 neutralizing antibody (anti-HMGB1) inhibited mechanical allodynia. Immunostaining data showed that compared with db/+ and C57 mice (postnatal 4 months), glial fibrillary acidic protein (GFAP) staining was significantly increased in the spinal cord of db/db mice. Anti-HMGB1 could effectively decrease GFAP expression. Real-time PCR showed that in postnatal 4 months, db/db mice induced significant increases of TNF-alpha, IL-1ß, IL-6 and monocyte chemoattractant protein-1 (MCP-1) in the spinal dorsal horn, while anti-HMGB1 (50 µg) effectively inhibited the up-regulation of these inflammatory mediators. Our results indicate that HMGB1 is significantly up-regulated in the spinal cord of type 2 diabetes, and inhibiting HMGB1 may provide a novel treatment for diabetic pain.