Guizhi-Shaoyao-Zhimu decoction attenuates monosodium urate crystal-induced inflammation through inactivation of NF-κB and NLRP3 inflammasome.

ETHNOPHARMACOLOGICAL RELEVANCE: Guizhi-Shaoyao-Zhimu decoction (GSZD), a classical traditional Chinese medicine (TCM) prescription, is used empirically to treat various types of arthritis in TCM clinical practice. However, the underlying mechanisms of GSZD on gouty inflammation are not totally elucidated. AIM OF STUDY: The purpose of this study is to investigate the effects of GSZD on peritoneal recruitment of neutrophils, production of proinflammatory mediators, activations of nuclear factor (NF)-κB and nucleotide oligomerization domain-like receptor protein-3 (NLRP3) inflammasome in mice with monosodium urate crystal (MSU)-induced peritonitis (MIP). MATERIALS AND METHODS: Mice were intragastrically administered with GSZD for 7 days. After the last administration, mice were intraperitoneally injected with MSU. Peritoneal exudates of mice were harvested, and total peritoneal cells were calculated. Levels of interleukin (IL)-1ß, IL-6 and monocyte chemotactic protein (MCP)-1 in peritoneal exudates were tested by enzyme-linked immunosorbent assay. Expressions of IL-1ß, NLRP3, cysteinyl aspartate specific proteinase (caspase)-1, apoptosis-associated speck-like protein containing the caspase activation and recruitment domain (ASC), phosphorylated (p)-p65, inhibitor of NF-κB (IκB)α, p-IκB kinase (IKK)ß, nuclear p65, p-mitogen-activated protein kinases (MAPKs) in peritoneal cells were analyzed by Western blot. Binding activity of NF-κB to DNA was measured by a Trans AM™ kit for p65. Interaction between ASC and pro-caspase-1 was assessed by co-immunoprecipitation assay. RESULTS: Total peritoneal cells, levels of IL-1ß, IL-6 and MCP-1 were significantly reduced by GSZD treatment in peritoneal exudates of MIP mice. As for the activation of NF-κB, GSZD treatment significantly reduced the levels of p-p65, p-IKKß, nuclear p65 and p-MAPKs, enhanced the level of IκBα and abated the binding ability of NF-κB to DNA in peritoneal cells of MIP mice. As for the activation of NLRP3 inflammasome, GSZD treatment significantly reduced the levels of IL-1ß, NLRP3 and caspase-1, and alleviated the interaction between ASC and pro-caspase-1 in peritoneal cells of MIP mice. Nevertheless, GSZD didn't remarkably change the level of ASC. CONCLUSIONS: These results suggest that GSZD attenuates the MSU-induced inflammation through inhibiting the activations of NF-κB and NLRP3 inflammasome.

[Modeling spatial distributions of hydrogen and oxygen stable isotopes in surface soils of the upper reaches of Minjiang River based on machine languages]. / åŸºäºŽæœºå™¨è¯­è¨€çš„å²·æ±Ÿä¸Šæ¸¸æµåŸŸè¡¨å±‚åœŸå£¤æ°¢æ°§ç¨³å®šåŒä½ç´ ç©ºé—´åˆ†å¸ƒæ¨¡æ‹Ÿ.

To study the feasibility of simulating the spatial distribution of hydrogen and oxygen stable isotopes composition (δ2H and δ18O) in the surface soil based on the machine learning method and to investigate large-scale distribution of δ2H and δ18O in the upper reaches of Minjiang River, 183 soil samples were collected from the 0-10 cm soil layer. After variable selection, back propagation (BP) neural network, random forests (RF) and support vector machine (SVM) were used to model the δ2H and δ18O of the study area, with the accuracies being evaluated. The structural equation model (SEM) was used to reveal the mechanism between the auxiliary variables and the δ2H and δ18O of soil water. The results showed that the RF model had the highest prediction accuracy, and could explain 75.0% and 64.0% of the variations of δ2H and δ18O in the surface soil, respectively. In this model, soil water content was the most important auxiliary variable, contributing 48.9% and 37.4% to δ2H and δ18O. Vegetation factors had stronger influence on δ2H and δ18O in the surface soil than climate factors, and the influence of climate factors on δ2H and δ18O was media-ted by vegetation factors. Among all the auxiliary variables, hydrogen/oxygen isotope of precipitation had the lowest effect on δ2H and δ18O due to the fractionation. The δ2H and δ18O in the surface soil of the upper reaches of the Minjiang River changed significantly across different months during the growing season. The increases of δ2H and δ18O in the early growing season and the decreases in the late growing season were mainly affected by vegetation, while climate change led to a small fluctuation in the middle growing season.

Simiao Wan attenuates monosodium urate crystal-induced arthritis in rats through contributing to macrophage M2 polarization.

ETHNOPHARMACOLOGICAL RELEVANCE: Simiao Wan (SMW) is a classical traditional Chinese medicine (TCM) prescription to empirically treat gouty arthritis (GA) in TCM clinical practice. However, the potential mechanisms of SMW on GA are not fully evaluated. AIM OF STUDY: The aim of this study is to investigate the role of macrophage polarization in the anti-GA activity of SMW. MATERIALS AND METHODS: Rats were intragastricly treated with SMW for consecutive 7 days. On day 6, monosodium urate (MSU) crystal-induced arthritis (MIA) in the ankle joint was prepared. Paw volume, gait score and histological score were measured. Levels of interleukin (IL)-1ß and IL-10 in serum were detected by enzyme-linked immunosorbent assay. Expressions of inducible nitric oxide synthase (iNOS), arginase (Arg)-1, phosphorylated (p)-p65, inhibitor of nuclear factor (NF)-κB (IκB)α, p-signal transducer and transcription activator (STAT)3 and p-Janus kinase (JAK)2 in synovial tissues were determined by Western blot. RESULTS: The elevated paw volume, gait score and histological score in MIA rats were significantly decreased by SMW treatment. Meanwhile, SMW significantly decreased the IL-1ß level and increased the IL-10 level in serum of MIA rats. Furthermore, SMW reduced the expressions of iNOS, p-p65 and enhanced the expressions of Arg-1, IκBα, p-STAT3 and p-JAK2 in synovial tissues of MIA rats. CONCLUSIONS: The results suggest that SMW attenuates the inflammation in MIA rats through promoting macrophage M2 polarization.

[Effect of Electroacupuncture Intervention on IL-4/STAT 6 and NF-κB p 65 Signaling in Cerebral Cortex in Focal Cerebral Ischemia/Reperfusion Injury Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on expression of cerebral cortical interleukin 4 (IL-4), signal transducer and activator of transcription 6 (STAT 6) and NF-κB family subunit p 65 in focal cerebral ischemia/reperfusion injury (CIRI) rats, so as to explore its mechanisms in reducing cerebral ischemia. METHODS: Forty-five male SD rats were randomly divided into sham group (n=15), CIRI model group (n=15), and EA group (n=15). The CIRI model was established by middle cerebral artery occlusion/reperfusion (MCAO/R). EA (1 mA, 2 Hz/100 Hz) was applied to "Baihui"(GV 20) and the left "Hegu" (LI 4)-"Taichong" (LR 3) for 20 min, once every 12 h, three times altogether. The expression levels of p-STAT 6, STAT 6 and NF-κB p 65 in the ischemic cortex were detected by immunohistochemistry and Western blot, separately, and the content of IL-4 was detected by ELISA. RESULTS: Compared with the sham group, cerebral IL-4 content and the expression levels of NF-κB p 65 immunoactivity and protein were significantly up-regulated in the CIRI model group (P<0.001, P<0.01). Following EA intervention, the levels of IL-4, p-STAT 6 expression and ratio of p-STAT 6/STAT 6 were notably increased in the EA group than in the model group (P<0.01),while the levels of cerebral NF-κB p 65 immunoactivity and protein expression were significantly down-regulated (P<0.01,P<0.05). CONCLUSIONS: EA of GV 20 and LI 4-LR 3 can up-regulate cerebral cortical IL-4 content and p-STAT 6 expression, and down-regulate NF-κB p 65 expression in CIRI rats, which may contribute to its effect in improving cerebral ischemia by reducing inflammatory injury.

Xitong Wan attenuates inflammation development through inhibiting the activation of nuclear factor-κB in rats with adjuvant-induced arthritis.

ETHNOPHARMACOLOGICAL RELEVANCE: Xitong Wan (XTW), a traditional Chinese herbs formula, has been used to treat "Bi Zheng" in the clinical practice of traditional Chinese medicine (TCM) for hundreds of years. However, no scientific validation is available on the anti-rheumatic effect of XTW. AIM OF STUDY: This study was carried out to investigate the effects of XTW on joints swelling, joints destruction, production of inflammatory mediators and nuclear factor-κB (NF-κB) activation in rats with adjuvant-induced arthritis (AIA). MATERIALS AND METHODS: AIA was induced by intradermal injection of Complete Freund's adjuvant in the footpad of Wistar rats. Paw volume was measured every 7 days during XTW treatment. Histological score was calculated by hematoxylin and eosin staining. Osteoclast number in articular tissues was counted by tartrate-resistant acid phosphatase staining. Levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 in serum were detected by enzyme-linked immunosorbent assay. Levels of NF-κBp65 and inhibitor of NF-κB (IκB)α in synovium were analyzed by Western blot assay. RESULTS: Compared with AIA group rats, XTW significantly decreased the paw volume of AIA rats. Meanwhile, XTW significantly reduced the histological score and osteoclast number in articular tissues of AIA rats. In addition, XTW markedly abated the levels of TNF-α, IL-1ß and IL-6 in serum, as well as enhanced the level of IκBα in synovium of AIA rats. However, XTW did not show significant effect on the level of p65 in synovium of AIA rats. CONCLUSIONS: These results suggest that XTW attenuates the inflammation development through inhibiting the NF-κB-mediated proinflammatory cytokines production in AIA rats. Our study provides the scientific evidence of XTW on treatment of rheumatoid arthritis in the clinical practice of TCM.

[Effects of Minocycline on the Neuronal Plasticity in Rats after Focal Cerebral Ischemia Reperfusion Injury].

OBJECTIVE: To observe the effects of minocycline on morphology and the expression of synaptophysin in cortical tissues of rats after cerebral ischemia reperfusion injury. METHODS: 36 male SD rats were randomly divided into 3 groups: sham group, model group [ischemia reperfusion (I/R)] and minocycline (Min) group (treated with minocycline for 14 d, 3 mg/kg, 2 times/d ). Middle cerebral artery occlusion (MCAO) was used to established as focal cerebral I/R model. At 14 d after I/R. Neurological functional recovery was evaluated using the staircase test, the cell morphology in cortex was evaluated by HE staining, the neurite growth was observed by immunostaining with anti-microtubule-associated protein-2 (MAP-2) antibody, the expression of synaptophysin in pei-infarct region was tested by Western blot. RESULTS: In the sham group, the rats did not show any neurological deficits. The neurons in the cortex were arranged in neat rows and the morphology were normal, the MAP-2 positive neurons showed longer neuronal processes than the model group. Compared to the model group, minocycline significantly improved forelimb motor function, increased the expression of synaptophysin and the number of MAP-2-positive cells in peri-infarct region (P < 0.05). CONCLUSION: Minocycline could improve the neurite regrowth and the expression of synaptophysin of neuron in ischemic cortex, promote neurological functional recovery of rats after MCAO, which is related to regulate the neuronal plasticity.

[Influence of electroacupuncture intervention on hippocampal IL-1beta content and I kappa B kinase beta expression in focal cerebral ischemia/reperfusion rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Baihui" (GV 20) and left "Hegu" (LI 4)-"Taichong" (LR 3) on levels of hippocampal IL-1beta content and I kappa B kinase (IKK) beta protein in focal cerebral ischemia/reperfusion (CI/R) injury rats so as to reveal its mechanism underlying improvement of cerebral ischemia. METHODS: One hundred and ninety-eight male SD rats were randomly divided into sham group (n = 54), model group (n = 72) and EA group (n = 72) which were further randomized into 12 h, 24 h and 48 h subgroups according to the duration of CI/R. The focal CI/R injury model was established by middle cerebral artery occlusion (MCAO) and reperfusion. EA (1 mA, 2 Hz/100 Hz) was applied to "Baihui" (GV 20) and the left "Hegu" (LI 4)-"Taichong" (LR 3) for 20 min, once every 12 h. The level of IL-1beta content in the right hippocampus tissue was detected by ELISA and the expression levels of hippocampal IKKbeta protein were detected by immunohistochemistry and Western blot, respectively. RESULTS: Compared with the 3 subgroups of the sham group, the levels of hippocampal IL-1beta contents at time-points of 12 h, 24 h and 48 h after MCAO were significantly increased in the model group (P < 0.01). In comparison with the model group, the contents of hippocampal IL-beta at the 3 corresponding time-points in the EA group were significantly reduced (P < 0.01). The expression levels of hippocampal IKKbeta protein at the 3 time-points after MCAO shown by immunohistochemistry and Western blot analysis were significantly higher in the model group than in the sham group (P < 0.05), and considerably lower in the EA group than in the model group (P < 0.05). CONCLUSION: EA of GV 20 and LI 4-LR 3 can obviously reduce CI/R injury induced increase of IL-1beta content and IKKbeta expression level in the hippocampus, which may be responsible for its effect in alleviating cerebral infarction by reducing inflammatory injury in the hippocampus.

Electroacupuncture Could Regulate the NF-κB Signaling Pathway to Ameliorate the Inflammatory Injury in Focal Cerebral Ischemia/Reperfusion Model Rats.

The activated nuclear factor-KappaB signaling pathway plays a critical role in inducing inflammatory injury. It has been reported that electroacupuncture could be an effective anti-inflammatory treatment. We aimed to explore the complex mechanism by which EA inhibits the activation of the NF- κ B signal pathway and ameliorate inflammatory injury in the short term; the effects of NEMO Binding Domain peptide for this purpose were compared. Focal cerebral I/R was induced by middle cerebral artery occlusion for 2 hrs. Total 380 male Sprague-Dawley rats are in the study. The neurobehavioral scores, infarction volumes, and the levels of IL-1 ß and IL-13 were detected. NF- κ B p65, I κ B α , IKK α , and IKK ß were analyzed and the ability of NF- κ B binding DNA was investigated. The EA treatment and the NBD peptide treatment both reduced infarct size, improved neurological scores, and regulated the levels of IL-1 ß and IL-13. The treatment reduced the expression of IKK α and IKK ß and altered the expression of NF- κ B p65 and I κ B α in the cytoplasm and nucleus; the activity of NF- κ B was effectively reduced. We conclude that EA treatment might interfere with the process of NF- κ B nuclear translocation. And it also could suppress the activity of NF- κ B signaling pathway to ameliorate the inflammatory injury after focal cerebral ischemia/reperfusion.

[Effects of electroacupuncture on endothelial progenitor cells in peripheral blood and bone marrow in local cerebral ischemia/reperfusion rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) of "Hegu" (LI 4) on blood stromal cell-derived factor 1alpha (SDF-1alpha), CXC chemokine receptor 4+ (CXCR 4+) positive cells and endothelial progenitor cells (EPCs) and bone marrow (BM) EPCs levels in local cerebral ischemia/reperfusion injury (CI/IR) rats, so as to study its mechanisms underlying improvement of cerebral ischemia. METHODS: A total of 54 SD rats were randomly divided into normal control, model, and EA groups. The latter two groups were further divided into 1 d, 2 d, 3 d and 7 d subgroups (4 time-points), respectively. CI/RI model was established by occlusion of the middle cerebral artery. EA (40 Hz/60 Hz, 1-2 mA) was applied to bilateral "Hegu" (LI 4) for 15 min, once daily, 1, 2, 3 and 7 days, respectively. Flow cytometer was used to detect the counts of EPCs and CXCR 4+ cells in the peripheral blood, and BM EPCs. Serum SDF-1alpha was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with the normal control group, the percentages of blood EPCs and CXCR 4+ cells and BM EPCs, and serum SDF-1alpha content on the 1st day, and the percentage of blood CXCR 4+ cells on the 2nd day were increased significantly in the model group (P<0.01, P<0.05). In comparison with the model group, percentages of blood EPCs and CXCR 4+ cells on the 1st day of EA group were reduced significantly (P<0.01), but the percentage of blood EPCs on the 2nd day, those of blood CXCR 4+ cells on the 2nd day and 3rd day, serum SDF-1alpha content on the 2nd day, and the percentage of BM EPCs on the 2nd day in the EA group were all up-regulated significantly (P<0.05, P<0.01). No significant differences were found among the 3 groups in blood EPCs percentages on the 3rd day and 7th day, in blood SDF-1alpha content and BM EPCs percentage on the 7th day (P>0.05). CONCLUSION: Generally, 2 days' EA of LI 4 can up-regulate the percentages of blood and BM EPCs and blood CXCR 4+ cells as well as blood SDF-1alpha protein content in CI/RI rats, which may contribute to its effect in improving cerebral ischemia.

[Combination of acupuncture, cupping and medicine for treatment of fibromyalgia syndrome: a multi-central randomized controlled trial].

OBJECTIVE: To evaluate the clinical effect of combination of acupuncture, cupping and medicine for treatment of fibromyalgia syndrome. METHODS: By using multi-central randomized controlled method, 186 cases were randomly divided into an acupuncture combined with cupping and western medicine group (group A), an acupuncture combined with cupping group (group B) and a western medicine group (group C) and treated continuously for 4 weeks. The treatment of acupuncture combined with cupping was produced by acupuncture at five mental points and moving cupping on the Hechelu of the back, once evrey other day, thrice each week, and the western medicine therapy by oral administration of Amitriptyline, once each day. The scores of McGill Pain Questionnaire (MPQ), the amount of tenderness point and the time of producing effect were compared and the therapeutic effects were assessed with the Hamilton Depression Scale (HAMD). RESULTS: The cured and markedly effective rate was 65.0% (39/60) in the group A, which was superior to 15.9% (10/63) in the group B and 16.1% (9/56) in the group C (both P < 0.001). After treatment, the scores of MPQ and HAMD and the amount of tenderness point all decreased in the three groups, group A being significantly better than group B and group C, and the time of producing effect in the group A was more earlier than those in the group B and the group C. CONCLUSION: The therapeutic effect of combination of acupuncture, cupping and medicine on fibromyalgia syndrome is superior to that of the simple acupuncture combined with cupping or the simple medicine.