IL-1 Receptor Antagonist mRNA Research Articles

Anti-Inflammatory Therapy for Temporomandibular Joint Osteoarthritis Using mRNA Medicine Encoding Interleukin-1 Receptor Antagonist

Messenger RNA (mRNA) is an emerging drug modality for protein replacement therapy. As mRNA efficiently provides protein expression in post-mitotic cells without the risk of insertional mutagenesis, direct delivery of mRNA can be applied, not only as an alternative to gene therapy, but also for various common diseases such as osteoarthritis (OA). In this study, using an mRNA-encoding interleukin-1 receptor antagonist (IL-1Ra), we attempted anti-inflammatory therapy in a rat model of the temporomandibular joint (TMJ) OA, which causes long-lasting joint pain with chronic inflammation. For the intra-articular injection of mRNA, a polyplex nanomicelle, our original polymer-based carrier, was used to offer the advantage of excellent tissue penetration with few immunogenic responses. While the protein expression was transient, a single administration of IL-1Ra mRNA provided sustained pain relief and an inhibitory effect on OA progression for 4 weeks. The mRNA-loaded nanomicelles provided the encoded protein diffusely in the disc and articular cartilage without upregulation of the expression levels of the pro-inflammatory cytokines IL-6 and tumor necrosis factor-α (TNF-α). This proof-of-concept study demonstrates how anti-inflammatory proteins delivered by mRNA delivery using a polyplex nanomicelle could act to alleviate OA, stimulating the development of mRNA therapeutics.

Upregulation of IL-1 Receptor Antagonist by Aspirin in Glial Cells via Peroxisome Proliferator-Activated Receptor-Alpha.

Background:Neuroinflammation is a recognized aspect of Alzheimer’s disease (AD) and other neurological illnesses. Interleukin 1 receptor antagonist (IL-1Ra) is an anti-inflammatory molecule, which inhibits inflammatory molecules in different cells including brain cells. However, mechanisms for upregulating IL-1Ra in brain cells are poorly understood.Objective:Since aspirin is a widely available pain reliever that shows promise beyond its known pain-relieving capacity, we examined whether aspirin could upregulate the IL-1Ra in the brain.Methods:We employed PCR, real-time PCR, western blot, immunostaining, chromatin immunoprecipitation (ChIP), and lentiviral transduction in glial cells. 5xFAD mice, an animal model of AD, were treated with aspirin orally via gavage.Results:Aspirin increased the expression of IL-1Ra mRNA and protein in primary mouse astrocytes and mouse BV-2 microglial cells. While investigating the mechanism, we found that the IL-1Ra gene promoter harbors peroxisome proliferator response element (PPRE) and that aspirin upregulated IL-1Ra in astrocytes isolated from peroxisome proliferator-activated receptor-beta knockout (PPARβ–/–), but not PPARα–/–, mice. Moreover, we observed that aspirin bound to tyrosine 314 residue of PPARα to stimulate IL-1Ra and that aspirin treatment also increased the recruitment of PPARα to the IL-1Ra promoter. Accordingly, aspirin increased IL-1Ra in vivo in the brain of wild type and PPARβ–/–, but not in PPARα–/– mice. Similarly, aspirin treatment also increased astroglial and microglial IL-1Ra in the cortex of 5xFAD, but not 5xFAD/PPARα–/– mice.Conclusion:Aspirin may reduce the severity of different neurological conditions by upregulating IL-1Ra and reducing the inflammation.

Effect of Interleukin 1 Receptor Antagonist Gene on Stable Expression Bone Marrow Mesenchymal Stem Cells and Early Aseptic Loosening of Hip Prosthesis of Mouse.

The research aimed to investigate the diagnostic value of Interleukin 1 receptor antagonist (IL-1Ra) in the early aseptic loosening of hip prosthesis and whether IL-1Ra can be expressed in bone marrow mesenchymal stem cells. In this study, the IL-1Ra gene was firstly connected to the lentiviral vector LV5, and the lentiviral vector LV5-home-IL1Ra was obtained by recombination. Then the recombinant LV5-home-IL1Ra was co-transfected with the virus-assisted plasmid into 293T cells and packaged to produce lentivirus. Bone marrow-derived stem cells (BMSCs) were infected with packaged lentiviruses. The relative expression of IL-1Ra mRNA in BMSCs was detected by fluorescence quantitative PCR. The expression of IL-1Ra protein in BMSCs was detected by western blot transfer electrophoresis. Peripheral venous blood samples from 108 patients and healthy subjects underwent total hip replacement were collected to detect the levels of plasma biomarkers procollagen type I carboxy-terminal propeptide (PICP), N-telopeptide cross-links of type I collagen (NTX), osteoprotegerin (OPG), TNGα, receptor activator of NF-kappaB ligand (RANKL), and IL-1β. The recombinant lentivirus vector IL-1Ra was successfully constructed by 2% agarose gel electrophoresis. Lentivirus-mediated IL-1Ra gene could efficiently transfection bone marrow mesenchymal stem cells, and the cell growth density reached about 80% at 72h after infection. The transfection rate was about 90%, and the fluorescence was enhanced. The relative mRNA and protein expression levels of IL-1Ra in the BMSCs-IL-1Ra group were significantly higher than those in the BMSCs group and the BMSCs-con group (P < 0.01). The late loosening group of IL-1β was significantly higher than the stable prosthesis group and the healthy group (P < 0.05). The ROC curve showed that IL-1 background had strong diagnostic sensitivity and specificity, which was similar to the X-ray score of osteolysis and had the most significant diagnostic significance. Lentivirus-transfected exogenous IL-1Ra can be expressed stably in mouse bone marrow mesenchymal stem cells, and IL-1β, an antagonist of IL-1Ra, plays an important role in the early aseptic loosening of hip prosthesis.

312 Effects of castration stress on behaviors and leukocyte cytokine gene expression in Korean cattle bull calves

Abstract This study investigated behavioral, physiological, and inflammatory responses, as well as leukocyte cytokine gene expression, of Korean cattle calves following surgical castration. Nineteen Korean cattle bull calves (average body weight, 254.5 kg; average age, 8.2 months) were assigned to one of two treatment groups: control (n = 9) and surgical castration (n = 10). Castration was performed surgically using Newberry knives and a Henderson castrating tool. Blood was collected immediately before castration and at 0.5 h, 6 h, 1 d, 3 d, 7 d, and 14 d after castration, and analyzed cortisol and substance P concentrations and leukocyte cytokine gene expression by quantitative real-time PCR. Behaviors were observed for 3 h, from 0.5 to 3.5 h after castration. Feed intake was recorded daily, and body weight was measured 1 d prior to the experiment and 14 d after castration. Castration decreased average daily gain (P = 0.005) and gain-to-feed ratio (P = 0.003). Castration reduced the time spent eating (P &amp;lt; 0.001) and the frequency of eating (P = 0.003) and increased (P &amp;lt; 0.001) the frequency of lying during the 3 h after castration. Castration temporarily increased circulating plasma cortisol (P &amp;lt; 0.001) and salivary cortisol concentrations (P = 0.03) at 0.5 h after castration. Castration temporarily increased (P &amp;lt; 0.05) plasma substance P concentration at 1 d after castration. Castration increased plasma haptoglobin concentration at 1 d and 3 d after castration. With regard to leukocytes, castration increased (P &amp;lt; 0.05) mRNA levels of inflammatory cytokine interleukin-1-beta and interleukin-1 receptor antagonist (IL1RA) genes at 6 h after castration, and increased (P &amp;lt; 0.05) IL1RA, interleukin-1-alpha, and interleukin-6 mRNA levels at 1 d after castration. In conclusion, castration of Korean cattle bull calves temporarily induced stress, retarded growth, and affected behaviors and inflammatory cytokine gene expression.

Infants with evolving bronchopulmonary dysplasia demonstrate monocyte-specific expression of IL-1 in tracheal aspirates.

Bronchopulmonary dysplasia (BPD) remains a devastating consequence of prematurity. Repeated inflammatory insults worsen lung injury, but there are no predictors for BPD-related respiratory outcomes or targeted therapies. We sought to understand inflammatory mechanisms in evolving BPD through molecular characterization of monocytes in tracheal aspirates from infants at risk for developing BPD. We performed flow cytometry targeting myeloid cell populations on prospectively collected tracheal aspirates from intubated patients born before 29 wk of gestation and <30 days old. We identified CD14+CD16+ (double-positive) and CD14+CD16- (single-positive) monocytes and characterized their gene expression profiles by RNA sequencing and quantitative PCR. We further analyzed differential gene expression between time points to evaluate changes in monocyte function over the first weeks of life. Expression of IL-1A, IL-1B, and IL-1 receptor antagonist mRNA was increased in monocytes collected at day of life (DOL) 7, DOL 14, and DOL 28 compared with those collected at DOL 3. This study suggests that early changes in monocyte-specific IL-1 cytokine pathways may be associated with evolving BPD.

Induction of M2 Macrophages Prevents Bone Loss in Murine Periodontitis Models

Periodontitis is characterized by the progressive destruction of tooth-supporting alveolar bone, which is mainly caused by chronic inflammation in response to persistent bacterial insult. It has recently become clear that the pathogenesis of periodontitis is associated with a high ratio of proinflammatory M1 (classically activated) macrophages to anti-inflammatory M2 (alternatively activated). To decrease the inflammatory activity, we locally delivered the C-C motif chemokine ligand 2 (CCL2) using controlled-release microparticles (MPs). CCL2 is known to promote chemotaxis of M0 or M2 phenotype macrophages to the inflamed site and induce M2 phenotype polarization locally. Our in vitro data showed that CCL2 increased the number of M2 phenotype macrophages, decreased TNF-α secretion, and enhanced chemotaxis of RAW264.7 cells toward CCL2 MPs. Moreover, we induced periodontal disease in 2 animal models through inoculation of Porphyromonas gingivalis and ligature around the murine molar. Micro–computed tomography analysis showed significant reduction of alveolar bone loss in the CCL2 MP treatment group when compared with a blank MP group and a no-treatment periodontitis group in both models. Immunohistologic analysis showed a significant increase in the M2 phenotype subset and a decrease in the M1 phenotype subset in the CCL2 MP group of the P. gingivalis–induced model. Also, in both models, tartrate-resistant acidic phosphatase staining showed significantly fewer numbers of osteoclasts in the CCL2 MP group in alveolar bone area. Moreover, quantitative polymerase chain reaction results showed a significant increase in IL-1RA (interleukin 1 receptor antagonist) mRNA expression and a decrease in RANKL (receptor activator of nuclear factor kappa-Β ligand) mRNA expression in the CCL2 MP group in the ligature model. In summary, manipulation of endogenous M2 phenotype macrophages with CCL2 MPs decreased the M1 phenotype:M2 phenotype ratio and prevented alveolar bone loss in mouse periodontitis models. The delivery of CCL2 MPs provides a novel approach to treat periodontal disease.

Phosphodiesterase 4B negatively regulates endotoxin-activated interleukin-1 receptor antagonist responses in macrophages

Activation of TLR4 by lipopolysaccharide (LPS) induces both pro-inflammatory and anti-inflammatory cytokine production in macrophages. Type 4 phosphodiesterases (PDE4) are key cAMP-hydrolyzing enzymes, and PDE4 inhibitors are considered as immunosuppressors to various inflammatory responses. We demonstrate here that PDE4 inhibitors enhance the anti-inflammatory cytokine interleukin-1 receptor antagonist (IL-1Ra) secretion in LPS-activated mouse peritoneal macrophages, and this response was regulated at the transcriptional level rather than an increased IL-1Ra mRNA stability. Studies with PDE4-deficient macrophages revealed that the IL-1Ra upregulation elicited by LPS alone is PKA-independent, whereas the rolipram-enhanced response was mediated by inhibition of only PDE4B, one of the three PDE4 isoforms expressed in macrophages, and it requires PKA but not Epac activity. However, both pathways activate CREB to induce IL-1Ra expression. PDE4B ablation also promoted STAT3 phosphorylation (Tyr705) to LPS stimulation, but this STAT3 activation is not entirely responsible for the IL-1Ra upregulation in PDE4B-deficient macrophages. In a model of LPS-induced sepsis, only PDE4B-deficient mice displayed an increased circulating IL-1Ra, suggesting a protective role of PDE4B inactivation in vivo. These findings demonstrate that PDE4B negatively modulates anti-inflammatory cytokine expression in innate immune cells, and selectively targeting PDE4B should retain the therapeutic benefits of nonselective PDE4 inhibitors.

Influence of melatonin on IL-1Ra gene and IL-1 expression in rats with liver ischemia reperfusion injury.

The aim of the present study was to explore the influence of melatonin (MT) on rats with liver ischemia reperfusion injury (IRI) and its mechanism. A total of 66 male Sprague-Dawley rats were randomly divided into 3 groups: i) Normal control group, ii) ischemia reperfusion group (IR group) and iii) melatonin treatment group (MT group). Rats in the MT group were administered an intraperitoneal injection of MT (10 mg/kg, 1 ml) at 70 and 35 min before ischemia, early reperfusion, and 1 and 2 h after reperfusion, respectively. Blood was removed at the normal time-point (prior to any processes), 35 min before ischemia, 2, 4, 8 and 24 h after reperfusion. Subsequently the rats were sacrificed. The pathological changes of liver tissues, interleukin-1 receptor antagonist (IL-1Ra) gene and IL-1 expression levels were detected. There were no evident differences between the immediate reperfusion and 2 h IR group and MT group. The liver structure injury of the 4, 8 and 24 h MT groups were improved to various differences compared to the corresponding IR group; liver IL-1β of the MT group at 35 min after ischemia, and 2, 4, 8 and 24 h after reperfusion was evidently lower than that of the IR group (P<0.05); IL-1Ra mRNA expression in the 2 h MT group was higher compared to the 2 h IR group by 4.85-fold; and IL-1Ra mRNA expression in the 4 h MT group was higher compared to the 4 h IR group by 9.34-fold. Differences between two groups at other time-points were <2-fold. In conclusion, MT can upregulate IL-1Ra gene expression by reducing generation of IL-1 thus reducing IRI.

IMQ-induced skin inflammation in mice is dependent on IL-1R1 and MyD88 signaling but independent of the NLRP3 inflammasome.

The pathogenesis of inflammatory skin diseases such as psoriasis involves the release of numerous proinflammatory cytokines, including members of the IL-1 family. Here we report overexpression of IL-1α, IL-1β, and IL-1 receptor antagonist mRNA, associated to expression of IL-23p19, IL-17A, and IL-22 in skin cells, upon topical application of the TLR7 agonist imiquimod (IMQ) in C57BL/6J mice. IMQ-induced skin inflammation was partially reduced in mice deficient for both IL-1α/IL-1β or for IL-1 receptor type 1 (IL-1R1), but not in IL-1α- or IL-1β-deficient mice, demonstrating the redundant activity of IL-1α and IL-1β for skin inflammation. NLRP3 or apoptosis-associated Speck-like protein containing a Caspase recruitment domain-deficient mice had no significant reduction of skin inflammation in response to IMQ treatment, mainly due to the redundancy of IL-1α. However, IMQ-induced skin inflammation was abolished in the absence of MyD88, the adaptor protein shared by IL-1R and TLR signaling pathways. These results are consistent with the TLR7 dependence of IMQ-induced skin inflammation. Thus, IL-1R1 contributes to the IMQ-induced skin inflammation, and disruption of MyD88 signaling completely abrogates this response.

IL-1 receptor antagonist improves morphine and buprenorphine efficacy in a rat neuropathic pain model

An interesting research and therapeutic problem is the reduced beneficial efficacy of opioids in the treatment of neuropathic pain. The present study sought to investigate the potential role of IL-1 family members in this phenomenon. We studied the time course of changes in IL-1alpha, IL-1beta, IL-1 receptor type I and IL-1 receptor antagonist mRNA and protein levels experienced by rats after chronic constriction injury (CCI) of the sciatic nerve using qRT-PCR and Western blot analysis. In CCI-exposed rats, spinal levels of IL-1alpha mRNA were slightly downregulated on the 7th day, and protein levels were not changed on the 7th and 14th days. Levels of IL-1 receptor antagonist and IL-1 receptor type I were slightly upregulated in the ipsilateral part of the spinal cord on the 7th and 14th days; however, protein levels were not changed at those time points. Interestingly, we observed that IL-1beta mRNA and protein levels were strongly elevated in the ipsilateral part of the dorsal spinal cord on the 7th and 14th days following CCI. Moreover, in rats exposed to a single intrathecal administration of an IL-1 receptor antagonist (100ng i.t.) on the 7th and 14th day following CCI, symptoms of neuropathic pain were attenuated, and the analgesic effects of morphine (2.5µg i.t.) and buprenorphine (2.5µg i.t.) were enhanced. In summary, restoration of the analgesic activity of morphine and buprenorphine by blockade of IL-1 signaling suggests that increased IL-1beta responses may account for the decreased analgesic efficacy of opioids observed in the treatment of neuropathy.

Neutrophils Play a Crucial Role in the Early Stage of Nonalcoholic Steatohepatitis via Neutrophil Elastase in Mice.

Neutrophils infiltration in liver is one of the typical histological characteristics of nonalcoholic steatohepatitis (NASH) in both animal models and human subjects. This study was aimed to investigate the role of neutrophils in the process of NASH and its underling mechanisms. C57BL/6J mice were fed with either standard chow (SC) or methionine/choline-deficient (MCD) diet for 1, 2, 4, 8weeks, respectively. C57BL/6J APOE(-/-) mice were fed with SC or high-fat high-cholesterol (HFHC) diet. Anti-Ly6G antibody was employed to deplete neutrophils and sivelestat was used to inhibit neutrophil elastase (NE). MCD-diet-receiving mice with neutrophil depletion had much lower serum ALT activity, liver inflammation, and mRNA levels of proinflammatory genes in the early stage of NASH (1 and 2weeks) when compared to non-neutrophil-depleted mice. NE inhibitor sivelestat could recapitulate the effects of neutrophil depletion in APOE(-/-) mice fed with HFHC diet. As the disease progressed (4 and 8weeks), neutrophil depletion did not lower serum ALT levels and liver lesions due to activation of Kupffer cells. Finally, we found neutrophils also affected anti-inflammation cytokine interleukin-1 receptor antagonist mRNA expression. Neutrophils play a crucial role in the early stage of NASH via NE.

Interleukin-1 receptor antagonist (IL1RN) is associated with suppression of early carcinogenic events in human oral malignancies.

Inflammatory abnormalities have been implicated in the pathogenesis of various human diseases, including cancer. Interleukin-1 receptor antagonist (IL1RN) is a potent anti-inflammatory molecule that modulates the biological activity of the proinflammatory cytokine, interleukin-1. The aim of this study was to examine the expression of IL1RN in oral squamous cell carcinomas (OSCCs), and to determine its clinical significance. Expression levels of IL1RN in matched normal and tumor specimens from 39 OSCCs were evaluated using real-time quantitative polymerase chain reaction methods, and immunohistochemical analysis. Protein expression of IL1RN was also examined in 18 oral premalignant lesions (OPLs). Expression of IL1RN mRNA was significantly downregulated in OSCCs compared with normal tissues. Decreased expression of IL1RN protein was also observed in OPLs and OSCCs. The IL1RN expression level was lower in the OPL cases with severe dysplasia compared to those with mild/moderate dysplasia. Significantly downregulated IL1RN expression was observed in all OSCC lesion sites examined when compared with the matched normal tissues. However, the decreased level of IL1RN expression did not correspond with tumor progression. Noteworthy, IL1RN expression was higher in the advanced OSCC cases (T3/T4) compared to early cases (T1/T2). Among OSCC samples, relatively higher IL1RN expression was associated with active tumor development in the OSCCs occurring in the buccal mucosa, oral floor, fauces and gingiva, but not the tongue. These data suggest that IL1RN may exhibit opposing characteristics in oral malignancies depending on the stage of cancer development, suppressing early carcinogenic events, yet promoting tumor development in some lesion sites. Thus, IL1RN could represent a reliable biomarker for the early diagnosis of OSCCs. Furthermore, IL1RN may possess unknown and complex functions in the developed OSCC.

Acute phase cytokines, TAC1, and toll-like receptor4 mRNA expression and health associated with group size in veal calves

Chronic stressors are a major health and well-being issue in animals. Immune status of animals under chronic stress is compromised, thus reducing disease resistance and compromising well-being of the animal. The objective of this study was to determine the influence of group size of veal calves on immune status and leukocyte mRNA expression of acute phase cytokines, toll-like receptor 4 (TLR4) and tachykinin 1 (TAC1) over a five-month finishing period. Holstein bull calves (n=168), 44±3 days of age were assigned to one of three treatments; 2, 4, or 8calves/pen (pen space allowance of 1.82m2/calf). Jugular blood samples were collected at the day of grouping and then monthly for 4 months. The differential leukocyte counts were determined and mRNA was extracted from the leukocytes. Reverse transcription-qPCR was used to measure the gene expression of interleukin-1 (IL-1β), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor (TNF-α), TLR4, and TAC1 in leukocytes. Health was evaluated before grouping and monthly for 4 months. On the 1st month after grouping, veal calves that were housed in groups of 8 have greater expression of IL-1β mRNA than calves housed in groups of 4 or 2 (treatment×month, P=0.04). Also at 1 month, groups of 8 had greater TAC1 expression (P<0.05) than calves housed in groups of 4 or 2. However, the expression of IL-1Ra, TNF-α, and TLR4 were not influenced by group size. In the first month of the trial, calves in groups of 8 coughed more (P<0.05) than calves in groups of 2 and coughed more than calves in groups of 4 and 2 during the 2nd month (treatment×month, P=0.03). Calves housed in groups of 8 tended to have greater neutrophil percentage (P=0.09), neutrophil to lymphocyte ratio (P=0.06), and had lower lymphocyte percentage (P=0.06) than those housed in groups of 4 or 2. In conclusion, the number of veal calves in a group, given the same space during the finishing period did not alter IL-1Ra, TNF-α, and TLR4 mRNA expression. However, housing of calves in groups of 8 was associated with greater expression of IL-1β and TAC1 mRNA in peripheral blood leukocytes, and coughing during the first 2 months after grouping. Therefore, housing of veal calves in larger groups may lead to greater susceptibility to respiratory disease and stress.

Effects of intrauterine growth retardation and postnatal high-fat diet on hepatic inflammatory response in pigs

The objective of this study was to investigate the glucose and insulin response of pigs with intrauterine growth retardation (IUGR) to a high-fat (HF) feeding regimen and to observe the underlying toll-like receptor-4 (TLR4) signalling pathway. Weaned piglets with IUGR or normal birthweight (NBW) (n = 20 each) received during the whole fattening period control diets (0% lard) or HF diets (HF, 10% lard). At about 110 kg body weight, pigs were euthanised to collect hepatic samples. Compared with NBW pigs, IUGR pigs had lower daily gain and feed intake. Growth rate of pigs was increased by HF feeding. Pigs fed HF diets had lower peak concentrations of glucose and insulin, which decreased more slowly than in pigs that received the control diets. The mRNA expression abundances of peroxisome proliferator-activated receptor coactivator-1α (PGC-1α), TLR4, myeloid differentiation factor 88 (MyD88), tumour necrosis factor-α (TNF-α), interleukin 6 (IL-6) and IL-1 receptor antagonist (IL-1Rn) were significantly affected by HF feeding, and IL-6 and IL-1Rn mRNA expressions were up-regulated in IUGR pigs compared with NBW pigs. Western blot analysis indicated that HF feeding elevated the protein expressions of TLR4 signalling pathway, as TLR4, MyD88, IκB kinase β and nuclear factor-κB, and insulin signalling-related proteins, as phosphorylated insulin receptor substrate-1 and phosphorylated protein kinase B. In summary, hepatic TLR4 signalling pathway and inflammatory response induced by HF feeding played an important role in the aggravated development of insulin resistance in pigs.

Thyroid hormone preconditioning alleviates reperfusion-induced renal inflammation in mice

Objective To investigate the influence of thyroid hormone T3 preconditioning on interleukin-10(IL-10) and interleukin-1 receptor antagonist(IL-1Ra) modulation and neutrophil infiltration after renal ishchemia/reperfusion(IR) in mice,so as to study the protective effect of T3 on IR kidney.Methods Totally 120 male C57BL/6 mice were randomly divided into four groups(n=30),namely,control group(sham operation),IR group(only received renal IR),T3+IR group(T3 preconditioning for 48 h before renal IR),and NaOH+IR group(received equivalent 0.1 mol/L NaOH soltuion 48 h before renal IR).The serum creatinine and blood urea nitrogen(BUN) were determined 24 h after reperfusion in each group;renal histological damages were scored using PAS staining;the levels of neutrophil infiltration were evaluated by MPO staining,and IL-10,IL-1Ra mRNA expression was examined by real-time PCR at 1,3,6,12,24,and 48 h after reperfusion.Results The serum creatinine and BUN levels of T3+IR group were significantly lower than those of IR group 24 h after reperfusion(P0.05),which was accompanied by lower histological score and significantly less neutrophil infiltration(P0.05).Real-time PCR results showed that IL-10 and IL-1Ra mRNA expression in T3+IR group was significantly higher than that in the IR group(P0.05) 12 h after reperfusion,which lasted for 48 h after reperfusion.The above parameters were similar between IR group and NaOH+IR group.Conclusion Thyroid hormone T3 preconditioning can alleviate renal IR injury,partly by increasing expression of IL-10 and IL-1Ra and subsequently reducing neutrophil infiltration at the late phase of renal IR.

The interleukin-1β system in the corpora lutea of pigs during early pregnancy and the estrous cycle

Expression of mRNAs encoding interleukin-1β (IL-1β), IL-1β receptor I (IL-1RI), IL-1 receptor accessory protein (IL-1RAcP) and IL-1 receptor antagonist (IL-1Ra), as well as synthesis of IL-1β and IL-1RI proteins, were examined in the corpus luteum (CL) during critical stages of CL activity on days 10–16 of pregnancy and 2–16 of the estrous cycle. Luteal cells were cultured in vitro with IL-1β, and the effect on release of steroid hormones was determined. Expression of the IL-1β system in the CL changed significantly during pregnancy and the estrous cycle. IL-1β, IL-1RI, and IL-1Ra mRNA levels were elevated on days 12–13, whereas IL-1RAcP mRNA was increased on days 15–16 of pregnancy. In cyclic CL, expression of IL-1β, IL-1RI, and IL-1RAcP mRNAs was increased on days 12–13. IL-1β and IL-1RI protein were highest in the CL on days 10–11 and 8–11 of pregnancy and the estrous cycle. Luteal cells harvested from gravid and cyclic CL produced IL-1β in vitro. IL-1β increased progesterone and estradiol-17β (E2) release by luteal cells on days 10–16 and 10–11 of pregnancy, respectively and on days 2–11 of the estrous cycle. IL-1β decreased the level of E2 produced by regressed CL (days 15–16). Expression of the IL-1β system in CL and IL-1β secretion from luteal cells changed depending on the status of the CL. These data show that IL-1β may be involved in intraluteal, luteotrophic regulation of CL functions in gravid and cyclic pigs.

Mucosal imbalance of interleukin-1β and interleukin-1 receptor antagonist in canine inflammatory bowel disease

Interleukin (IL)-1β is a key mediator of the inflammatory response. IL-1 receptor antagonist (IL-1Ra) regulates inflammation by functioning as an endogenous inhibitor of IL-1β. A disruption of the balance between IL-1β and IL-1Ra has been identified in human inflammatory bowel disease (IBD). The objective of this study was to determine whether there is an intestinal imbalance of IL-1β and IL-1Ra in canine IBD by comparing expression of IL-1β and IL-1Ra mRNA by real-time RT-PCR and expression of IL-1β and IL-1Ra protein by ELISA in 21 dogs with IBD, 15 dogs with intestinal lymphoma ('inflammatory' controls) and 20 healthy Beagles ('healthy' controls). A significant decrease in the intestinal IL-1Ra:IL-1β ratio of mRNA and protein was observed in IBD cases when compared with healthy control dogs. In contrast, a decrease in IL-1Ra:IL-1β ratio was not observed in dogs with intestinal lymphoma. The IL-1Ra:IL-1β protein ratio was negatively correlated with clinical severity in dogs with IBD. An intestinal imbalance between IL-1β and IL-1Ra production may play a role in the pathogenesis of canine IBD.

IL1RN genetic variations and risk of IPF: a meta-analysis and mRNA expression study

Idiopathic pulmonary fibrosis (IPF) is a rare and devastating lung disease of unknown aetiology. Genetic variations in the IL1RN gene, encoding the interleukin-1 receptor antagonist (IL-1Ra), have been associated with IPF susceptibility. Several studies investigated the variable number tandem repeat (VNTR) or single nucleotide polymorphisms rs408392, rs419598 and rs2637988, with variable results. The aim of this study was to elucidate the influence of polymorphisms in IL1RN on IPF susceptibility and mRNA expression. We performed a meta-analysis of the five case–control studies that investigated an IL1RN polymorphism in IPF in a Caucasian population. In addition, we investigated whether IL1RN mRNA expression was influenced by IL1RN polymorphisms. The VNTR, rs408392 and rs419598 were in tight linkage disequilibrium, with D′ > 0.99. Furthermore, rs2637988 was in linkage disequilibrium with the VNTR (D′ = 0.90). A haploblock of VNTR*2 and the minor alleles of rs408392and rs419598 was constructed. Meta-analysis revealed that this VNTR*2 haploblock is associated with IPF susceptibility both with an allelic model (odds ratio = 1.42, p = 0.002) and a carriership model (odds ratio = 1.60, p = 0.002). IL1RN mRNA expression was significantly influenced by rs2637988, with lower levels found in carriers of the (minor) GG genotype (p < 0.001). From this meta-analysis, we conclude that the VNTR*2 haploblock is associated with susceptibility to IPF. In addition, polymorphisms in IL1RN influence IL-1Ra mRNA expression, suggesting that lower levels of IL-1Ra predispose to developing IPF. Together these findings demonstrate that the cytokine IL-1Ra plays a role in IPF pathogenesis.

Energy-sensing Factors Coactivator Peroxisome Proliferator-activated Receptor γ Coactivator 1-α (PGC-1α) and AMP-activated Protein Kinase Control Expression of Inflammatory Mediators in Liver: INDUCTION OF INTERLEUKIN 1 RECEPTOR ANTAGONIST

Obesity and insulin resistance are associated with chronic, low grade inflammation. Moreover, regulation of energy metabolism and immunity are highly integrated. We hypothesized that energy-sensitive coactivator peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α) and AMP-activated protein kinase (AMPK) may modulate inflammatory gene expression in liver. Microarray analysis revealed that PGC-1α up-regulated expression of several cytokines and cytokine receptors, including interleukin 15 receptor α (IL15Rα) and, even more importantly, anti-inflammatory interleukin 1 receptor antagonist (IL1Rn). Overexpression of PGC-1α and induction of PGC-1α by fasting, physical exercise, glucagon, or cAMP was associated with increased IL1Rn mRNA and protein expression in hepatocytes. Knockdown of PGC-1α by siRNA down-regulated cAMP-induced expression of IL1Rn in mouse hepatocytes. Furthermore, knockdown of peroxisome proliferator-activated receptor α (PPARα) attenuated IL1Rn induction by PGC-1α. Overexpression of PGC-1α, at least partially through IL1Rn, suppressed interleukin 1β-induced expression of acute phase proteins, C-reactive protein, and haptoglobin. Fasting and exercise also induced IL15Rα expression, whereas glucagon and cAMP resulted in reduction in IL15Rα mRNA levels. Finally, AMPK activator metformin and adenoviral overexpression of AMPK up-regulated IL1Rn and down-regulated IL15Rα in primary hepatocytes. We conclude that PGC-1α and AMPK alter inflammatory gene expression in liver and thus integrate energy homeostasis and inflammation. Induction of IL1Rn by PGC-1α and AMPK may be involved in the beneficial effects of exercise and caloric restriction and putative anti-inflammatory effects of metformin.

도담탕합총명탕(導痰湯合聰明湯)이 LPS로 처리된 microglia 및 기억력 감퇴 생쥐 모델에 미치는 영향

Objectives : This experiment was designed to investigate the efficacy of DDTCMT hot water extract & ultra-fine powder on Alzheimer's Disease Model. Methods : The effects of the DDTCMT hot water extract on expression of IL-<TEX>$1{\beta}$</TEX>, IL-6, TNF-<TEX>${\alpha}$</TEX>, COX-2, NOS-II, IL-10, IL-1 receptor antagonist mRNA and production of IL-<TEX>$1{\beta}$</TEX>, IL-6, TNF-<TEX>${\alpha}$</TEX> in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. Expression of NO, ROS in BV2 microglial cell line treated by LPS and AChE activity in PC-12 cell treated by NGF were investigated. anti-AChE was observed through Western blot analysis. The effects of the DDTCMT hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine were investigated. Results : 1. The DDTCMT hot water extract significantly decreased the production of mIL-6, mNOS-II, mTNF-<TEX>${\alpha}$</TEX>, and increased the production of mIL-10, mIL-1 receptor antagonist. 2. The DDTCMT hot water extract significantly suppressed the production of IL-<TEX>$1{\beta}$</TEX>, IL-6, TNF-<TEX>${\alpha}$</TEX> in BV2 microglial cell line treated by LPS. 3. The DDTCMT hot water extract significantly suppressed the NO and ROS production in BV2 microglial cell line treated by LPS. 4. The DDTCMT hot water extract groups showed inhibition of AChE activity in NGF treated PC-12 cell line. 5. The DDTCMT hot water extract suppressed anti-AChE expression in NGF treated PC-12 cell line was observed by Western blot analysis. 6. The DDTCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine -induced impairment of memory in the experiment of Morris water maze. Conclusions : These results suggest that the DDTCMT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease.