Effect of Modified Buwangsan on Expression of Related Molecules in NLRP3 Inflammatory Signaling Pathway in Hippocampus of AD Rats / 中国实验方剂学杂志

Objective::To investigate the effects of modified Buwangsan on the learning and memory ability of Alzheimer's disease (AD) model rats and the expression of NOD-like receptor 3 (NLRP3), cysteine-containing aspartate-specific proteases 1 (Caspase-1) and interleukin-1 beta (IL-1β) in NLRP3 inflammatory pathway in hippocampus of AD model rats, and exploring the underlying mechanism of modified Buwangsan. Method::The 52 eligible rats were randomly divided into sham control group, AD model group, low-dose modified Buwangsan group (1.5 g·kg-1) and high-dose modified Buwangsan group (3 g·kg-1). AD mouse model was established by bilateral hippocampus injection of Aβ1-425 μL (2 g·L-1). The rats in low-dose and high-dose modified Buwangsan group received low and high dose modified Buwangsan respectively within the next 4 weeks, once daily. The learning and memory ability was tested by Morris water maze. The expression of NLRP3, Caspase-1 and IL-1β mRNA was tested by quantitative PCR(Real-time PCR) and Western blot. Result::As compared with the sham group, the learning and memory ability of the rats were significantly impaired (P<0.05). Compared with AD model group, the learning and memory ability and the expression levels of NLRP3, Caspase-1, and IL-1β mRNA and protein were all no statistical differences in low-dose modified Buwangsan group, while the learning and memory ability of the rats were significantly improved and the expression of NLRP3, Caspase-1 and IL-1β mRNA in hippocampus of rats was significantly decreased in high-dose modified Buwangsan group (P<0.05). Conclusion::High-dose modified Buwangsan could attenuate neuroinflammation in the hippocampus of AD mouse model via inhibiting the expression of NLRP3, Caspase-1 and IL-1β, which may be the mechanisms of modified Buwangsan could be used to ameliorate the learning and memory ability of AD mouse model.

Acupuncture combined with granule for nerve-root type cervical spondylosis and its effects on IL-6、TNF-α、IL-1β and hemorheological indexes / 中国针灸

OBJECTIVE@#To explore the clinical efficacy of acupuncture combined with granule for nerve-root type cervical spondylosis and its effects on serum interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and hemorheological indexes.@*METHODS@#A total of 114 patients with nerve-root type cervical spondylosis were randomly divided into an observation group and a control group, 57 cases in each group. The patients in both groups were treated with traction. The patients in the control group were treated with oral administration of granule, 4 g each time, 3 times a day, while based on the treatment of control group, the patients in the observation group were treated with acupuncture at Dazhui (GV 14), Tianzhu (BL 10), Houxi (SI 3), cervical Jiaji (EX-B 2), Quchi (LI 11), Hegu (LI 4) and Waiguan (TE 5), once a day. Both groups were treated for 4 weeks. The simplified McGill pain questionnaire (MPQ), neck disability index (NDI), numbness score, levels of IL-6, TNF-α, IL-1β in serum and hemorheological indexes were observed before and after treatment, and the clinical efficacy was compared between the two groups.@*RESULTS@#The total effective rate was 91.2% (52/57) in the observation group, which was higher than 71.9% (41/57) in the control group (<0.05). Compared before treatment, the scores of MPQ, NDI and numbness in the two groups were reduced after treatment (<0.05). After treatment, the scores of MPQ, NDI and numbness in the observation group were lower than those in the control group (<0.05). After treatment, the serum levels of IL-6, TNF-α and IL-1β in the two groups were reduced (<0.05), and those in the observation group were lower than the control group (<0.05). After treatment, the plasma viscosity, fibrinogen, low shear rate of whole blood viscosity and high shear rate of whole blood viscosity in the two groups were lower than before treatment (<0.05), and those in the observation group were lower than the control group (<0.05).@*CONCLUSION@#Acupuncture combined with granule have significant clinical efficacy for nerve-root type cervical spondylosis, which could reduce the serum levels of IL-6, TNF-α and IL-1β and improve hemorheology.

Pyrin Domain (PYD)-containing Inflammasome in Innate Immunity

Inflammasome is a cytosolic multiprotein complex to activate caspase-1 leading to the subsequent processing of inactive pro-interleukin-1-beta (Pro-IL-1beta) into its active interleukin-1 beta (IL-1beta) in response to pathogen- or danger-associated molecular pattern. In recent years, a huge progress has been made to identify inflammasome component as a molecular platform to recruit and activate caspase-1. Nucleotide-binding oligomerization domain-like receptor (NLR) family proteins such as NLRP1, NLRP3 or interleukin-1beta-converting enzyme (ICE)-protease activating factor (IPAF) have been first characterized to form inflammasome complex to induce caspase-1 activation. More recently, non-NLR type, pyrin-domain (PYD)-containing proteins such as pyrin or absent in melanoma2 (AIM2) were also proposed to form caspase-1-activating inflammasome machinery with apoptosis-associated speck-like protein containing a CARD (ASC), an essential adaptor molecule. Inflammasome pathways were shown to be crucial for protecting host organisms against diverse pathogen infections, but accumulating evidences also suggest that excessive activation of inflammasome/caspase-1 might be related to the pathogenesis of inflammation-related diseases. Indeed, mutations in NLRP3 or pyrin are closely associated with autoinflammatory diseases such as familial Mediterranean fever (FMF) syndrome or Muckle-Wells syndrome (MWS), indicating that the regulation of caspase-1 activity by inflammasome is a central process in these hereditary inflammatory disorders. Here, recent advances on the molecular mechanism of caspase-1 activation by PYD-containing inflammasomes are summarized and discussed.

Evaluation of osteogenic activity of periosteal-derived cells treated with inflammatory cytokines

INTRODUCTION: Skeletal homeostasis is normally maintained by the stability between bone formation by osteoblasts and bone resorption by osteoclasts. However, the correlation between the inflammatory reaction and osteoblastic differentiation of cultured osteoprogenitor cells has not been fully investigated. This study examined the effects of inflammatory cytokines on the osteoblastic differentiation of cultured human periosteal-derived cells. MATERIALS AND METHODS: Periosteal-derived cells were obtained from the mandibular periosteum and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured in an osteogenic induction Dulbecco's modified Eagle's medium (DMEM) medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. In this culture medium, tumor necrosis factor (TNF)-alpha with different concentrations (0.1, 1, and 10 ng/mL) or interleukin (IL)-1beta with different concentrations (0.01, 0.1, and 1 ng/mL) were added. RESULTS: Both TNF-alpha and IL-1beta stimulated alkaline phosphatase (ALP) expression in the periosteal-derived cells. TNF-alpha and IL-1beta increased the level of ALP expression in a dose-dependent manner. Both TNF-alpha and IL-1beta also increased the level of alizarin red S staining in a dose-dependent manner during osteoblastic differentiation of cultured human periosteal-derived cells. CONCLUSION: These results suggest that inflammatory cytokines TNF-alpha and IL-1beta can stimulate the osteoblastic activity of cultured human periosteal-derived cells.

The Central and Peripheral Production of Pro-inflammatory Cytokine, IL-1b after Immune and Stress Stimulation in Rats

Interleukin-1beta (IL-1beta), one of the pro-inflammatory cytokines, acts as an endogenous pyrogen and is an important mediator of behavioral and physiological responses to immune stimulation as well as exposure to stressors. The objective of the present study was to examine the pattern of central or peripheral IL-1beta response to lipopolysaccharide (LPS) or exposure to the foot shock stress (FS) in rats. After treatment of LPS (100microgram/kg) or exposure to the FS [ten times (0.8 mA) foot shocks for 5 sec each and 90 sec interval], body temperature and IL-1beta levels in plasma, spleen and brain were measured. Both LPS and FS stimuli elicited increased body temperature but showed different patterns of peripheral IL-1beta levels. LPS produced a widespread increase in IL-1beta levels in the plasma, spleen and brain, whereas FS produced a significant increase in IL-1beta levels only in the brain regions but not in plasma and spleen. The present study suggests that IL-1beta is, centrally or peripherally in different patterns, regulated by immune stimulation or exposure to stressors and IL-1beta plays an important role in mediating responses of sickness-like behaviors induced by immune stimuli or stressors.

The Effects of Air-borne Particulate Matters on the Alveolar Macrophages for the TNF-alpha and IL-1beta Secretion / 결핵및호흡기질환

BACKGROUND: PM is known to induce various pulmonary diseases, including asthma, cancer, fibrosis and chronic bronchitis. Despite the epidemiological evidence the pathogenesis of PM-related pulmonary diseases is unclear. METHODS: This study examined the effects of PM exposure on the secretion of TNF-alpha and IL-1beta in the cultured alveolar macrophages. The cultured primary alveolar macrophages were treated with the medium, PM (5~20 microgram/cm2), LPS (5ng/ml), and PM with LPS for 24h and 48h respectively. ELISA was used to assay the secreted TNF-alpha and IL-beta in the culture medium. Western blotting was used to identify and determine the level of proteins isolated from the culture cells. The cells cultured in the Lab-Tek(R) chamber slides were stained with immunocytochemical stains. RESULTS: PM induced TNF-alpha and IL-1beta secretion in the culturing alveolar macrophages, collected from the SPF and inflammatory rats. However, the effects were only dose-dependent in the inflammatory macrophages. When the cells were co-treated with PM and LPS, there was a significant synergistic effect compared with the LPS in the both cell types. CONCLUSION: PM might be play an important role in the induction and/or potentiation of various lung diseases by oversecretion of TNF-alpha and IL-1beta.

The Regulation of Cyclooxygenase-2 Expressionby Interleukin-1beta in WISH cells / 대한산부인과학회잡지

OBJECTIVE: To determine of the regulation of cyclooxygenase-2 (COX-2) expression by Interleukin-1beta in WISH cells. METHODS: Amnion WISH cells were incubated in media containing increasing concentrations of IL-1beta or with various inhibitors. Increased COX-2 expression was determined by Western blot analysis with anti-COX-2 antibody. Concomitant measurements of culture media PGE2 were made by an enzyme immunoassay. RESULTS: The COX-2 and prostaglandin E2 production induced by IL-1beta increased in a dose- and time-dependent manner. One of the regulating factors that induced COX-2 by IL-1beta was protein kinase C (PKC). PKC inhibitor, Ro 31-8220 was pretreated and continued treating by IL-1beta. Then, PKC inhibitor completely blocked COX-2 protein induction by IL-1beta. In contrast, COX-2 induction by IL-1beta after pretreating PKC stimulator, phobol 12-myristate 13-acetate was potentiated with synergism. Another factor in controlling COX-2 protein induction was identified as phosphatidylinositol 3-kinase (PI 3K). COX-2 protein induction by IL-1beta after pretreating PI 3K inhibitors, wortmannin and LY294002 strongly increased. This kind of result reflected that PI 3K act as negative regulator. COX-2 induction by IL-1beta was known to be regulated in not only transcription step, but also translation step after performing experiment of actinomycin and cycloheximide treatment. CONCLUSION: COX-2 protein and prostaglandin E2 production induced by IL-1beta were controlled by many factors in amnion cell. Among those factors, PKC and PI 3K have an important role, but their control mechanism act as positive and negative, respectively.

Reasonable usage of ectogenous glucocorticoids for the management of severely scalded rats / 第三军医大学学报

Objective To investigate the effects of different doses of dexamethasone used at different time on the serumal concentrations of TNF ?, IL 1? and IL 6 of rats after severe scald in order to provide experimental basis for reasonable usage of ectogenous glucocorticoids for severe traumas. Methods The serumal concentrations of TNF ?, IL 1? and IL 6 were detected by ELISA after intraperitoneal injection of different doses of dexamethasone into rats at different time after severe scald. Results High dose(5 mg/kg) and low dose (0.5 mg/kg) of dexathemasone used during the period from 4 h to 12 h after severe scald could both significantly reduce the serumal concentrations of TNF ?, IL 1? and IL 6, and the effect of high dose was much better than that of low dose. Both doses of dexathemasone used at 24 h after scald had a significant effect on IL 1? and IL 6, but there was no difference between the two doses. No obvious effects were found of high/low dose of dexamethasone used 48 h later after scald. Conclusion According to the anti inflammatory effect, the best therapeutic efficacy may result from high dose of ectogenous glucocorticoids during the period from 4 h to 12 h and low dose from 12 h to 24 h after severe scald.