Interference effect of Radix notoginseng on the expressions of Toll-like receptor 2/4 and NOD like receptor P1/3 in renal tissues of septic rats with acute kidney injury / 中国中西医结合急救杂志

Objective To investigate the expressions of Toll-like receptors and NOD-like receptor (TLR/NLRP) in septic rats with acute kidney injury (AKI) and influence of Radix notoginseng. Methods One hundred and fifty male Sprague-Dawley (SD) rats were selected, and they were divided into three groups: sham operation group, model group and Radix notoginseng pretreatment group with 50 rats in each group. Sepsis rats with AKI models were established by cecal ligation puncture (CLP); only was laparotomy performed in the sham operation group without ligation. The rats in Radix notoginseng pretreatment group were given Radix notoginseng (3 g/kg) for consecutive 3 days by gastric perfusion before treatment, and the sham operation group and sepsis model group rats were given equal amount of normal saline by gastric perfusion. After their blood was collected, at each following time points 6, 12 and 24 hours after modeling, 10 rats in each group were sacrificed, and the kidney specimens were collected. The endotoxin levels were detected by limulus assay; the levels of serum creatinine (SCr) and urea nitrogen (BUN) were detected by automatic biochemical analyzer; serum levels of interleukins (IL-1β, IL-18, IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA); the expression levels of NLRP1/3 and TLR2/4 mRNA and protein in renal tissue were detected by real-time fluorescence quantitative polymerase chain reaction (qPCR) and Western Blot, respectively; the pathological changes of renal tissues were observed by optical microscope. The 4-day and 7-day survival rates were observed in the remaining 20 rats in each group. Results The levels of endotoxin, SCr, BUN, IL-1β, IL-6, IL-18, and TNF-α and the expressions of NLRP1/3, TLR2/4 mRNA and proteins in sepsis model group and Radix notoginseng pretreatment group were significantly higher than those in sham operation group, and the levels of endotoxin, IL-18, and the expressions of NLRP1/3, TLR2/4 mRNA and proteins showed statistically significant differences at 6-hour after operation [endotoxin (kU/L): 61.3±25.7, 56.9±18.6 vs. 0.2±0.1, IL-18 (ng/L): 16.7±5.0, 13.8±2.9 vs. 10.6±2.8, NLRP3 mRNA (2-ΔΔCt): 6.3±1.9, 4.0±1.2 vs. 1.1±0.4, TLR mRNA (2-ΔΔCt): 5.7±1.3, 2.0±0.8 vs. 0.9±0.3, TLR4 mRNA (2-ΔΔCt): 4.4±1.2, 2.3±0.7 vs. 0.6±0.2, NLRP3/β-actin: 38.2±9.3, 26.1±7.2 vs. 18.3±5.1, TLR4/β-actin: 21.9±6.1, 16.2±4.4 vs. 10.9±2.8, TLR4/β-actin: 18.3±6.7, 12.0±3.9 vs. 7.5±2.0, all P < 0.05], the levels of SCr, BUN, IL-1β, TNF-α and IL-6 showed statistically significant differences at 12-hour after operation [SCr (μmol/L): 62.3±21.6, 38.1±13.9 vs. 36.0±11.9, BUN (mmol/L): 16.5±7.2, 6.9±2.6 vs. 6.8±2.5, IL-1β (ng/L): 37.6±10.9, 31.2±9.3 vs. 20.3±6.5, TNF-α (ng/L): 15.6±3.9, 10.2±2.8 vs. 7.3±2.1, IL-6 (ng/L):9.3±2.5, 6.8±1.7 vs. 5.0±1.3, all P < 0.05], the levels of expressions of NLRP3 mRNA and protein were obviously lower than those in sham operation group, and there were statistical significant differences immediately after 6-hour after operation [NLRP1 mRNA (2-ΔΔCt): 0.5±0.1, 0.8±0.2 vs. 1.6±0.5, NLRP3/β-actin: 8.0±2.1, 16.8±5.0 vs. 35.6±10.5, all P < 0.05], and the amplitude changes of the above indexes in Radix notoginseng pretreatment group were obviously smaller than those in sepsis model group (all P < 0.05); the survival rates of 4-day and 7-day in sepsis model group were significantly lower than those in sham operation group [4-day: 25% (5/20) vs. 95% (19/20), 7-day: 15% (3/20) vs. 95% (19/20), both P < 0.05], while the survival rate in Radix notoginseng pretreatment group was significantly higher than that in sepsis model group [respectively 65% (13/20) vs. 25% (5/20), 60% (12/20) vs. 15% (3/20), both P < 0.05]. Conclusions TLR2/4 and NLRP1/3 may be involved in the pathogenesis of AKI of septic rats. Traditional Chinese medicine Radix notoginseng possibly via regulating the expressions of TLR2/4 and NLRP1/3 can reduce the inflammatory response, in turn ameliorate kidney injury in septic rats and improve their renal functions.

The negative-pressure wound therapy promotes wound healing by enhancing angiogenesis in granulation tissue and wound healing through suppression of NLRX1 through via the up-regulation of miR-195expression by suppressing NLRX1 expression upregulation / 中国医师杂志

Objective To investigate the expression and the relationship with angiogenesis of miR-195 and NLR family member X1 (NLRX1) in granulation tissue after negative-pressure wound treatment (NPWT).Methods Six patients were collected who received negative pressure treatment with refractory wound granulation.The levels of miR-195, NLRX1 mRNA and NLRX1 proteins were measured.The expression of NLRX1 and the micro-vascular density (MVD) of CD31 were detected by immunohistochemistry (IHC).Results MiR-195 and MVD were significantly higher in granulation tissue after 7 days negative pressure treatment (P＜0.05), and NLRX1 was significantly lower (P ＜0.05).In granulation tissue,the expression of miR-195 was negatively correlated with NLRX1 (r =-0.856, P ＜0.001), the expression of NLRX1 was negatively correlated with MVD (r =-0.618, P ＜0.05), and the expression of miR-195 was positively correlated with MVD (r =0.630, P ＜ 0.05).Conclusions Negative pressure wound therapy can promote the formation of granulation vessels and the wound healing.The therapeutic mechanism may inhibit the expression of NLRX1 and upregulate the expression of miR-195 to promote angiogenesis.

NLRP12 and its role in occurrence and development of diseases / 国际儿科学杂志

NLRP12 is a member of the NOD-like receptors family in innate immune system,and plays an important role on resistance to intracellular pathogens.Once being activated,NLRP12 could recruit and combine with its downstream effector proteins into inflammasome,thus involving in pathogen clearance.Meanwhile,NLRP12 is one of the several anti-inflammatory molecules in the NOD-like receptors family.In this review,its structure,tissue distribution,function and association with diseases were discussed.

<b>“Mi-byo” and Homeostatic Inflammation : Possible Regulation by Herbal Medicine</b> / 日本東洋医学雑誌

“Mi-byo” (pre-disease) is a special concept in Chinese Medicine proposed about 2,200 years ago in the ancient text <i>Ko-tei-nai-kei</i>, which states that “a Saint-like Doctor” can cure “mi-byo”. However, no one has been saintly enough to explain an actual “mi-byo” status to date. In the 21 th century, as immunology has developed, the novel notion of “homeostatic inflammation” began to be postulated. Here, “homeostatic inflammation” means the self-repairing steps initiated by innate immune sensors when they encounter either PAMPs (pathogen-associated molecular patterns) or with DAMPs (danger signal-or damage-associated molecular patterns) composed of either lipids-or nucleic acids-related substances through their own TLRs (toll-like receptors) or NLRs (NOD-like receptors), respectively. If such “homeostatic inflammation” does correlate with the “mi-byo”, perhaps we can control it by using herbal medicines containing various saponins, essential oils, alkaroids, and flavonoids that may reinforce innate barriers by regulating the effect of lipids and nucleic acids.

NOD-Like Receptors in Infection, Immunity, and Diseases

Nucleotide-binding and oligomerization domain (NOD)-like receptors (NLRs) are pattern-recognition receptors similar to toll-like receptors (TLRs). While TLRs are transmembrane receptors, NLRs are cytoplasmic receptors that play a crucial role in the innate immune response by recognizing pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs). Based on their N-terminal domain, NLRs are divided into four subfamilies: NLRA, NLRB, NLRC, and NLRP. NLRs can also be divided into four broad functional categories: inflammasome assembly, signaling transduction, transcription activation, and autophagy. In addition to recognizing PAMPs and DAMPs, NLRs act as a key regulator of apoptosis and early development. Therefore, there are significant associations between NLRs and various diseases related to infection and immunity. NLR studies have recently begun to unveil the roles of NLRs in diseases such as gout, cryopyrin-associated periodic fever syndromes, and Crohn's disease. As these new associations between NRLs and diseases may improve our understanding of disease pathogenesis and lead to new approaches for the prevention and treatment of such diseases, NLRs are becoming increasingly relevant to clinicians. In this review, we provide a concise overview of NLRs and their role in infection, immunity, and disease, particularly from clinical perspectives.

Regulation of Toll-like receptors in fungal keratitis / 中华实验眼科杂志

Fungal keratitis (FK) ,a potential blinding disease,has been difficult to treat due to the limited number of approved antifungal drugs and the taxing dosing regimen.Toll-like receptors (TLRs) function as the pattern recognition receptors (PRRs) in mammals and play an essential role in the recognition of fungal components.There is a great amount of evidence that TLRs initiate the innate immunity in the FK.Furthermore, TLRs also play roles in shaping fungal-specific humoral and cellular adaptive immune responses.This review described the recent advances in interaction between TLRs and non-TLRs signal transduction,negative regulator and function of TLRs in corneal fungal infection.

Transcriptional Analysis of Nod-Like Receptors in a Mouse Model of Experimental Autoimmune Uveitis

PURPOSE: To evaluate the transcription pattern of Nod-like receptors (NLRs), the intracellular sensors, to detect danger signals in murine eyes with experimental autoimmune uveitis (EAU). METHODS: EAU was induced in B6 (C57BL/6) mice by subcutaneous injection of human interphotoreceptor retinoid binding protein and intraperitoneal injection of pertussis toxin. At 1, 2, and 3 weeks post-immunization, the eyeballs were extracted and subjected to histological and molecular assays using real-time reverse transcription polymerase chain reaction. RESULTS: The levels of nucleotide-binding oligomerization domain, Leucine rich Repeat and Pyrin domain 1 (NLRP1), NLRP3, nucleotide-binding oligomerization domain-containing protein 1 (NOD1), and NOD2 transcripts were increased at 2 weeks and gradually reduced thereafter. Notably, NLRP3 showed the highest expression in the eyes with EAU. Similarly, the transcript level of pro-inflammatory cytokine, interleukin-1beta, increased and reached a peak at 2 weeks post-immunization. The retinal structure was severely damaged by inflammation at 3 weeks post-immunization. CONCLUSIONS: Among NLRs, NLRP3 may induce inflammation in eyes after EAU immunization.

La estimulación de TLR, receptores tipo NOD y dectina-1 en neutrófilos humanos induce la producción de citocinas proinflamatorias / Stimulation of TLRs, Nod-like receptors and Dectin-1 in neutrophils induces the production of proinflammatory cytokines

Objetivo: evaluar la expresión y la función de receptores de reconocimiento de patrones como los de tipo Toll y los de tipo NOD, RIG-I/MDA5, la dectina-1 y moléculas adaptadoras, en neutrófilos humanos. Métodos: a partir de sangre periférica de individuos sanos se purificaron y cultivaron neutrófilos en el medio RMPI-1640, en presencia o ausencia de los agonistas específicos de los receptores de interés. La expresión de los receptores de reconocimiento de patrones se determinó por RT-PCR y la secreción de citocinas proinflamatorias, por ELISA. Resultados: los neutrófilos expresan un amplio espectro de receptores de reconocimiento de patrones y de moléculas adaptadoras. La estimulación de TLR4, TLR5, TLR7/8 induce la secreción de IL-1β e IL-6; la activación de la dectina-1 induce una alta producción de TNF-α, pero bajos niveles de IL-1β e IL-6. Conclusión: los neutrófilos expresan un amplio número de receptores de reconocimiento de patrones y su activación lleva a la expresión de diferentes citocinas proinflamatorias.

Objective: To evaluate the expression and function of pattern recognition receptors such as Toll-like receptors, RIG-I/MDA5, NOD-like receptors, Dectin-1 and adaptor proteins, in human neutrophils. Methods: Neutrophils from peripheral blood of healthy individuals were purified and cultured in RPMI-1640, in the presence or absence of specific agonists of the receptor of interest. The expression of pattern recognition receptors was determined by RT-PCR and the secretion of proinflammatory cytokines, by ELISA. Results: We observed that neutrophils express diverse patterns recognition receptors and adaptor molecules. Stimulation of TLR4, TLR5 and TLR7/8 induces the production of IL-1β and IL-6, and activation of Dectin-1 leads to secretion of high levels of TNF-α, but low levels of IL-1β and IL-6. Conclusion: Neutrophils express a large number of pattern recognition receptors and their activation leads to the expression of proinflammatory cytokines.

Toll-like Receptors and NOD-like Receptors in Innate Immune Defense during Pathogenic Infection

In response to invading pathogens, the body immune system develops an immediate defense mechanism, i.e., innate immune response, which is detected in almost all living organisms including mammals, plants, insects, etc. Recent studies have identified numerous innate immune receptors that are able to recognize pathogen-associated molecular patterns and transduce the essential intracellular signaling cascades to mount early and successful host defenses against infectious challenge. Among innate immune receptors, we will focus on two important receptors, toll-like receptors (TLRs) and nucleotide binding oligomerization domain (Nod)-like receptors, and their major intracellular signaling pathways that culminate to activate innate immune effectors and inflammatory mediators during pathogen infection. In this review, we address the recent advances of understanding intracellular signaling mechanisms by which TLRs and NLRs activate host immune defense and inflammation. The role and regulatory mechanisms by which a subet of NLRs-associated inflammasome activation induce interleukin-1beta secretion and their relevance with host defense will be also discussed. Both TLR- and NLR-mediated intracellular signaling networks serve crucial roles in mounting resistance to bacterial and viral infection through synthesis of immune mediators and antimicrobial chemicals during infection.

Autophagy as an Innate Immune Modulator

Autophagy is a fundamental cellular process in eukaryotic cells for maintaining homeostasis by degrading cellular proteins and organelles. Recently, the roles of autophagy have been expanded to immune systems, which in turn modulate innate immune responses. More specifically, autophagy acts as a direct effector for protection against pathogens, as well as a modulator of pathogen recognition and downstream signaling in innate immune responses. In addition, autophagy controls autoimmunity and inflammatory disorders by negative regulation of immune signaling. In this review, we focus on recent advances in the role of autophagy in innate immune systems.

Functional and Mechanistic Integration of Infection and the Metabolic Syndrome / 당뇨병

The metabolic syndrome refers to a well defined group of risk factors, including central obesity and inflammation, for the development of diabetes and cardiovascular disease. Interestingly, many studies have recently led to the emergence of somewhat unexpected relationships between several infectious diseases and various aspects of the metabolic syndrome. Our understanding of the mechanisms underlying these interactions is also rapidly developing and some of these are summarized in this article. We will focus first on bacterial infection, and most notably the role of gut microbiota in regulaton of both obesity and inflammation. In particular, we focus on the role of inflammasomes and propose that understanding the role of Toll-like receptors and Nod-like receptors in the pathogenesis of inflammatory disorders with or without infection may provide novel targets for prevention and/or treatment of associated diseases. Secondly, chronic bacterial or viral infection and emerging links with metabolism will be reviewed. Finally, consideratons of biomarkers for metabolic syndrome, in particular lipocalin-2, and their link with infection will be discussed.