Effect of Gegen Qinliantang on TLR4/IKKβ/NF-κB Pathway in KKAy Mice with Diabetes / 中国实验方剂学杂志

ObjectiveTo observe the effect of classical prescription Gegen Qinliantang（GGQLT） on inflammatory factors and key targets in the inflammatory pathways mediated by lipopolysaccharide in KKAy mice and explore its mechanism in improving spontaneous type 2 diabetes mellitus （T2DM）. MethodSixty-five SPF KKAy mice with spontaneous T2DM and 13 C57BL/6J mice （control） were selected in the barrier system and fed on a high-fat diet. The model was properly induced in 44 mice in the context of random blood glucose exceeding or equal to 13.9 mmol·L-1. Then the mice were assigned into a normal group （20 mL∙kg-1 normal saline）， a model group （20 mL∙kg-1 normal saline）， an acarbose group （3.9 mg∙kg-1）， and high- and low-dose GGQLT groups （1.82 and 0.45 g∙kg-1）， with 11 mice in each group. The mice in each group were treated correspondingly by gavage for eight weeks， once per day. Blood glucose and body weight were systematically evaluated. Twelve hours after the last administration， blood samples were collected from the eyes， and the serum and muscle and liver tissues were extracted. The levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and glucose transporter type 4 （GluT4） were detected by semi-quantitative enzyme-linked immunosorbent assay （ELISA）. The protein expression of IκB kinase β （IKKβ） and nuclear factor-κB （NF-κB） in muscle tissues and Toll-like receptor 4 （TLR4） in liver tissues was detected by Western blot. ResultCompared with the normal group， the model group showed increased body weight and blood glucose （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced body weight and blood glucose （P<0.05， P<0.01）. As revealed by ELISA results， compared with the normal group， the model group showed increased levels of TNF-α and IL-6 （P<0.01） and deceased GluT4 level （P<0.05）. Compared with the model group， the groups with drug treatment showed reduced levels of TNF-α and IL-6 （P<0.05， P<0.01）， and the acarbose group and the high-dose GGQLT group showed increased GluT4 level （P<0.05， P<0.01）. As displayed by Western blot results， compared with the normal group， the model group showed increased protein expression of IKKβ， NF-κB， and TLR4 （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced protein expression of IKKβ， NF-κB， and TLR4 （P<0.05， P<0.01）. ConclusionGGQLT can inhibit the inflammatory cascade effect and improve T2DM by down-regulating the levels of key inflammatory factors in the TLR4 pathway， inhibiting their activation， and increasing the translocation and activity of GluT4 on the basis of the regulation of intestinal flora.

Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway / 中国实验方剂学杂志

Objective:To observe the intervention of phlegm-stasis co-treatment on the myocardial Toll-like receptor 4 （TLR4）/nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）/nuclear factor-<italic>κ</italic>B inhibitor （I<italic>κ</italic>B） signaling pathway， and to investigate its mechanism in improving myocardial inflammation in rats with diabetes mellitus （DM）. Method:Forty-five male SD rats of SPF grade were randomly divided into a normal group， a phlegm-resolving （Xiao Xianxiongtang， 4.05 g·kg^-1） group， a stasis-resolving （Xuefu Zhuyutang， 7.02 g·kg^-1） group， a co-treatment （Didang Xianxiong decoction， 8.10 g·kg^-1） group， an alagebrium chloride （3 mg·kg^-1） group， and a model group. Except for normal group， the other rats was induced by a single intraperitoneal injection of 55 mg·kg^-1streptozotocin （STZ） to establish DM model. After adaptive feeding for three weeks， the rats were treated correspondingly by gavage daily for eight weeks. Rats were sampled under anesthesia. Enzyme-linked immunosorbent assay（ELISA） was used to detect the protein expression of TLR4 and tumor necrosis factor-alpha （TNF-<italic>α</italic>） in myocardial tissues. The expression levels of NF-<italic>κ</italic>B p65 and I<italic>κ</italic>B<italic>α</italic> were detected by immunohistochemistry. NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， TNF-<italic>α</italic>， and TLR4 mRNA expression levels were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）. Result:The protein and mRNA levels of TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α </italic>were higher in the model group than those in the normal group （<italic>P</italic><0.01）. TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α</italic> protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group （<italic>P</italic><0.01）. The inter-group comparison revealed that the co-treatment group showed more manifest reduction in protein and mRNA expression levels of TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α，</italic> and TNF-<italic>α </italic>than the phlegm-resolving group and the stasis-resolving group （<italic>P</italic><0.05<italic>，P</italic><0.01）. Conclusion:The co-treatment of phlegm and stasis can improve myocardial inflammation in DM rats， with superior effect to either the phlegm-resolving method or the stasis-resolving method. The underlying mechanism may be related to the inhibition of TLR4/NF-<italic>κ</italic>B/I<italic>κ</italic>B signaling pathway activation.

Ginkgetin alleviates LPS-induced inflammatory damage in macrophages of mouse model via inhibiting IκB-α degradation / 中国药理学通报

Aim To explore the anti-inflammatory effect of natural compound Ginkgetin on lipopolysaccharide-induced mouse primary peritoneal macrophages and the underlying mechanism, in order to provide a theoretical basis for the development of clinical drug candidates. Methods MTT test kit was used to detect the cytotoxicity of Ginkgetin on mouse primary peritoneal macrophages; ELISA and RT-qPCR methods were used to detect the anti-inflammatory effect of different concentrations of Ginkgetin on LPS-induced cell inflammation injury model; Western blot was used to detect the anti-inflammatory mechanism of ginkgo flavonoids. Results Compared with LPS stimulation group, Ginkgetin treatment group produced a concentration-dependent anti-inflammatory effect, which could be attributed to the fact that Ginkgetin could inhibit LPS-induced activation of NF-κB signaling pathway. MTT results also showed that ginkgo flavonoids had little toxicity to mouse primary peritoneal macrophages. Conclusions Ginkgelin alleviates LPS-induced inflammatory injury of mouse primary peritoneal macrophages by inhibiting the activation of NF-κB signaling pathway. It is expected to be a natural monomer antiinflammatory drug candidate.

Tanshinone IIA inhibits LPS induced inflammation of RAW264.7 cells by regulating TLR4/IκB-α/NF-κB signaling pathway / 中国药理学通报

Aim To establishan inflammatory model of mouse monocyte macrophages ( RAW264. 7) using li-popolysaccharide (LPS), and to investigate the antiinflammatory activity and mechanism of tanshinone II-A (Tan IIA). Methods Cell viability was determined by CCK-8 method. Cell migration was detected by Tr-answell apparatus. TNF-α, IL-6, IL-1 p, MCP-1 content in cell supernatant was analyzed using ELISA method. The protein expression of MMP-2, MMP-9, TLR4, κB-α, p-κB-α, NFκB and p-NFκB in RAW264.7 cells was investigated by Western blot. Results Tan IIA significantly inhibited .the secretion of TNF-α, IL-6, IL-1β and MCP-1 in LPS induced RAW264.7 cell culture medium , and significantly down-regulated the expression of matrix metalloprotein-ase-2 (MMP-2), MMP-9, TLR4, p-κB-α and p-NFκB , inhibited IκB-α phosphorylation and NFκB entry into the nucleus and activation. Conclusion Tan IIA can inhibit the release of inflammatory factors through the regulation of TLR4/IκB-α/NFκB signaling pathway.

Effect of Modified Huangqi Biejiatang Combined with Auricular Acupressure on Diabetic Peripheral Neuropathy of Qi-Yin Deficiency Syndrome and Serum MyD88/IκB Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the effects of modified Huangqi Biejiatang combined with auricular acupressure on diabetic peripheral neuropathy （DPN） due to Qi and Yin deficiency and serum myeloid differentiation factor 88/inhibitor of nuclear factor-<italic>κ</italic>B （MyD88/I<italic>κ</italic>B） signaling pathway. Method:One hundred and forty cases were randomly divided into an observation group （<italic>n</italic>=70） and a control group （<italic>n</italic>=70）. In addition to routine treatments like dietary intervention and the regulation of fasting blood glucose （FBG） and blood pressure， the modified Huangqi Biejiatang combined with auricular acupressure was further provided in the observation group， while mecobalamine was administered in the control group. After four-week intervention， the toronto clinical scoring system （TCSS） score， traditional Chinese medicine （TCM） syndrome score， the conduction velocities of motor and sensory nerves （median nerve， common peroneal nerve， tibial nerve， and ulnar nerve）， glucose metabolism indexes ［fasting plasma glucose （FPG）， 2 h postprandial blood glucose （2 h PG）， and hemoglobin A1c （HbA1c）］， intestinal genera （<italic>Clostridium</italic>， <italic>Prauserella</italic>， <italic>Bacteroides</italic>， and <italic>Faecalibacterium</italic>）， as well as the serum MyD88， I<italic>κ</italic>B<italic>α</italic>， and phosphorylated I<italic>κ</italic>B<italic>α </italic>（p-I<italic>κ</italic>B<italic>α</italic>） levels in the MyD88/I<italic>κ</italic>B signaling pathway before and after treatment were observed in the two groups， for comparing their clinical efficacy and safety. Result:The total effective rate of the observation group was 85.3% （58/68）， which was higher than 48.5% （32/66） of the control group （<italic>χ</italic>²=6.143， <italic>P</italic><0.05）. The comparison with the control group revealed that the scores of TCSS and TCM syndrome， the levels of FPG， 2 h PG， HbA1c， MyD88， and p-I<italic>κ</italic>B<italic>α</italic>， as well as the abundances of <italic>Clostridium</italic> and <italic>Prauserella</italic> in the observation group were decreased （<italic>P</italic><0.05）， while the conduction velocities of motor and sensory nerves （median nerve， common peroneal nerve， tibial nerve， and ulnar nerve） were significantly accelerated （<italic>P</italic><0.05）. Besides， the abundances of <italic>Bacteroides</italic> and <italic>Faecalibacterium</italic> and I<italic>κ</italic>B<italic>α</italic> level were significantly elevated （<italic>P</italic><0.05）. The incidence of adverse reactions in the observation group was 1.5% （1/68）， lower than 12.1% （8/66） in the control group （<italic>χ</italic>²=4.328， <italic>P</italic><0.05）. Conclusion:The modified Huangqi Biejiatang combined with auricular acupressure alleviates DPN due to Qi and Yin deficiency， which may be attributed to the regulation of serum MyD88/I<italic>κ</italic>B signaling pathway.

Schisandrin C alleviates atherosclerosis and inflammation in ApoE-/-mice fed with arterial high-fat diet / 中国药理学通报

Aim To investigate the effects of schisandrin C on arterial inflammation and atherosclerosis of ApoE-/-mice fed with high-fat diet. Methods ApoE-/-mice were divided into high fat diet group (HFD) and high fat diet group + schisandrin group (HFD + Sch C). High fat diet and Sch C were given at the same time. After 12 weeks, mice were executed and arterial tissues were collected. RT-q PCR and Western blot were respectively used to detect the expressions of IL-6, TNF-α, ICAM-1, β-actin and the phosphorylation and expression of IκB-α in arteries. Oil red O staining and biochemical kit were respectively used to detect the lipid changes in aortic root and the blood lipid levels. Results Compared with HFD group, the area of atherosclerotic plaque in HFD + Sch C was reduced (P < 0. 05), but Sch C did not affect blood lipid levels. Compared with HFD group, the mRNA expression of TNF-α, IL-6 and ICAM-1 and the phosphorylation of IκB-α of arterial tissue in HFD + Sch Cgroup decreased (P < 0. 05). Conclusions Sch C could effectively alleviate atherosclerosis induced by high-fat diet in ApoE-/-mice, accompanied by alleviation of arterial inflammation.

Effect of Shenling Baizhusan on Protein and mRNA Expression of NF-κB p65，IκBα，IκKβ in Ulcerative Colitis Rats with Syndrome of Dampness Stagnancy Due to Spleen Deficiency / 中国实验方剂学杂志

Objective:To observe the effect of Shenling Baizhusan on the protein and mRNA expression of inhibitor of nuclear factor kappa B kinase (IκK)/inhibitor of nuclear factor kappa B(IκB)/nuclear factor kappa B(NF-κB) signaling pathway in the colon of rats with ulcerative colitis (UC) of spleen deficiency and dampness stagnation type, and to explore the mechanism of Shenling Baizhusan in the treatment of UC. Method:The 48 Wistar rats were randomly divided into normal group, model group, Shenling Baizhusan group (15.6 g·kg-1) and osalazine sodium group (0.68 g·kg-1), 12 rats in each group. The model of UC with spleen deficiency and dampness stagnation was reproduced by trinitrobenzene sulfonic acid (TNBS)/ethanol enema combined with environment and diet intervention.Serum levels of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β(IL-1β) were determined by enzyme-linked immunosorbent assay（ELISA）. The expression of NF-κB p65, IκBα, IκKβ protein in colon tissue was measured by Western blot and immunohistochemical method, and the mRNA expression of NF-κB p65, IκBα and IκKβ in colon tissue of rats in each group was detected and compared by real time polymerase chain reaction (Real-time PCR). Result:Compared with normal group,the levels of TNF-α,IL-6 and IL-1β in the serum, the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the model group was significantly higher than that of normal group (P<0.01), and the protein and mRNA expression of IκBα was significantly lower than that of normal group (P<0.01). Compared with model group,the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the Shenling Baizhusan group and osalazine sodium group were significantly decreased (P<0.01), and the protein and mRNA expression of IκBα was significantly increased (P<0.01). Conclusion:Shenling Baizhusan can obviously down regulate the protein and mRNA expression of NF-κB p65, IκKβ,up regulate the expression of IκBα in colon tissue of UC rats with spleen deficiency and dampness stagnation. The inhibition of IκK/IκB/NF-κB signal pathway activation by Shenling Baizhusan is an important mechanism of its role in protecting intestinal mucosa.

Effect of Xiao Chaihutang on NF-κB Signaling Pathway in Synovial Tissue of CFA Rats / 中国实验方剂学杂志

Objective:To observe the expression of tumor necrosis factor receptor-associated death domain (TARDD), nuclear transcription factor-κB inhibiting protein α(IκBα)IκB kinase-α (IKKα) and nuclear transcription factor (NF)-κB p65 protein in the NF-κB signaling pathway of synovial tissues of complete Freund's adjuvant (CFA) rats after treatment with Xiao Chaihutang (XCHT). Method:In animal experiments, SPF health adult female Wistar rats were used to prepare the CFA animal model of rats with rheumatoid arthritis with Freund's complete adjuvant and cattle Ⅱ collagen type. According to the random number table, the rats were randomly divided into the normal group, the model group, the low-dose XCHT group, the medium-dose XCHT group, the high-dose XCHT group, and the Tripterygium glucosides group. The drugs were given at 7 d after the model was built. Both normal group and model group were given water for injection,and low-dose XCHT group(5.94 g·kg-1),medium-dose XCHT group(11.88 g·kg-1),high-dose XCHT group(23.76 g·kg-1),Tripterygium glucosides group(0.006 3 g·kg-1) were given corresponding drugs by gavage for three times a day, 2 mL/time. The histopathology of rat ankle joint was observed, and the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in the NF-κB signaling pathway in synovial tissue of CFA rats were detected by Western blot. Result:With the increase of the dosage of XCHT, the histopathological score of the right posterior ankle joint of the experimental rats was increased. And in the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in NF-κB signaling pathway in Synovial Tissue of CFA rats, compared with the model group, the statistical results of the low-dose XCHT group showed decreased protein expressions (PPPα, IκB α, NF-κB p65 in the NF-κB signaling pathway were significantly increased (PPα, IκBα, NF-κB p65 key protein expressions in the NF-κB signaling pathway and protein expressions in low-dose XCHT group were obviously lower (PPConclusion:This study shows that as the dose of Xiao Chaihutang increases, it could effectively improve synovitis, and suppress the expressions of key proteins in the inflammatory signaling pathway of NF-κB, thereby preventing inflammation and suppressing bone erosion.

Advance in Role of IκB Kinase beta in Inflammatory Response Mechanism post Spinal Cord Injury (review) / 中国康复理论与实践

IκB kinase beta (IKKbeta) is a key kinase in nuclear factor kappa B (NF-kappaB) signaling pathway. After spinal cord injury, IKKbeta is activated, and the signal pathway of NF-kappa B is abnormally activated, which produces a large number of inflammatory factors, which has a negative impact on the recovery of spinal cord injury. This article mainly summarized the structure and function of IKKbeta and its application in inflammatory reaction after spinal cord injury, trying to find a new target for the treatment of spinal cord injury.

Effect of Electroacupuncture on Inflammatory Response of Rats after Focal Cerebral Ischemia-reperfusion via Regulating IκB Kinases β Expression / 中国康复理论与实践

Objective:To observe the changes of inflammatory damage in the brain of rats after focal cerebral ischemia-reperfusion, and explore the effect of the initiation of IκB kinases β (IKKβ), which is the key protein of activating nuclear factor (NF)-kappa B signaling pathway in inflammatory response, and the mechanism of electroacupuncture inhibiting inflammatory damage. Methods:A total of 240 male Sprague-Dawley rats were randomly divided into sham group, ischemia-reperfusion group, electroacupuncture group, IKKβ silencing group, IKKβ overexpression group and IKKβ overexpression + electroacupuncture group, each group was further divided into six hours, twelve hours, 24 hours, 48 hours and 72 hours subgroups. The right middle cerebral artery occlusion reperfusion model was established by modified thread embolization. The IKKβ gene was intervened by gene silencing and gene overexpression technology. Results:Compared with the model group, the neurological function score increased (P < 0.05), the cerebral infarction volume decreased (P < 0.05), the activation of NF-κB p65 was inhibited, and the content of proinflammatory factors decreased (P < 0.05) in IKKβ silencing group. Compared with IKKβ silencing group, the above results were significantly worse in IKKβ overexpression group (P < 0.05), and microglia in cerebral ischemic cortex were significantly activated. The activation of microglia and activation of IKKβ were significantly inhibited in IKKβ overexpression + electroacupuncture group. Conclusion:IKKβ gene silencing could inhibit the inflammatory response of cerebral ischemic cortex mediated by NF-κB signaling pathway, and over-expression of IKKβ could lead to severe inflammatory damage in ischemic cortex. Electroacupuncture could inhibit the inflammatory damage after focal cerebral ischemia-reperfusion by regulating the activity of IKKβ.

Scalp-acupuncture Improves Neurological Function by Regulating Expression of NF-κB p 65 mRNA, IκB mRNA and IL-1 β and TNF-α in Parahippocampal Gyrus of Cerebral Ischemia Rats / 针刺研究

OBJECTIVE: To observe the effect of scalp-acupuncture intervention on the expression of parahippocampal factor-κB p 65 mRNA (NF-κB p 65 mRNA), IκB mRNA, interleukin-1 β (IL-1 β) and tumor necrosis factor-α (TNF-α) in rats with cerebral ischemia (CI), so as to investigate its molecular mechanisms underlying improving CI by reducing inflammatory response. METHODS: A total of 64 SD rats were randomized into normal control, model, medication and scalp-acupuncture groups, with 16 rats in each group. The focal CI model was established by middle cerebral artery occlusion (MCAO). Intraperitoneal injection of Pyrrolidine Dithiocarbamate (100 mg•kg－1•d－1) was administrated for rats in the medication group, once a day for 7 days. For rats of the scalp-acupuncture group, the acupuncture needles were rapidly inserted into bilateral Dingnieqianxiexian (MS 6) and Dingniehouxiexian (MS 7), followed by twirling the needles at 200 cycles/min for 1 min, once again every 10 min during 30 min's needle retention. The treatment was conducted once a day for 7 days. The neurologic deficit score (0－5 points, impaired consciousness, death, etc.) and neurological function score (motor, sensory and sensory tests, 0－10 points) were assessed according to Longa's (1989) and Schäbitz's (2004) methods, respectively. The expression levels of NF-κB p 65 mRNA and IκB mRNA in the parahippocampus gyrus tissue were detected by fluorescence quantitative reverse transcription-PCR, and IL-1 β and TNF-α proteins in the parahippocampus gyrus tissue were detected by immunohistochemistry. RESULTS: After modeling, the neurologic deficit and neurological function scores and the expression levels of NF-κB p 65 mRNA, IL-1 β and TNF-α in the parahip-pocampus were significantly increased in the model group than in the normal group (P<0.01), while the expression of IκB mRNA was considerably down-regulated (P<0.01). Following treatment intervention, the neurologic deficit and neurological function scores as well as NF-κB p 65 mRNA, and IL-1 β and TNF-α protein expression were significantly decreased in both scalp-acupuncture and medication groups compared with the model group (P<0.05, P<0.01), and the decreased expression of IκB mRNA was obviously increased (P<0.05)．. CONCLUSION: Scalp-acupuncture can improve neurologic function in cerebral ischemic rats, which is related with its effects in up-regulating the expression of IκB to inhibit the dissociation of NF-κB, then decreasing the expression of IL-1 β and TNF-α (reducing inflammatory response) in the parahippocampal gyrus tissue.

Breviscapine reduces intestinal ischemia reperfusion via inhibiting IκB-α/NF-κB/ICAM-1 pathway in a rat model / 实用医学杂志

Objective To investigate the effects of breviscapine injection on intestinal injury induced by intestine ischemia-reperfusion(IIR). Methods 48 males SD rats with 8-week old were randomly divided into 4 groups:Sham,intestine ischemia-reperfusion(IIR),EB+IIR,TP+IIR. Breviscapine injection 20 mg/(kg·d) was given intraperitoneally in EB + IIR group. TPCA-1(12 mg/kg)was given intravenously 30 min before surgery in TP+IIR group. Rats were subjected to superior mesenteric artery occlusion consisting of 45 min of ischemia and 6 h of reperfusion;sham laparotomy served as controls. Intestine pathology was assayed by H&E staining. Con-centrations of TNF-α,IL-1β,and IL-6 in intestinal mucosa were determined by ELISA. The protein expressions of IκB-α,NF-κB ,ICAM-1of intestine tissue were assayed by western blot. Results IIR induced serious intesti-nal injury ,evidenced as poor intestine pathology ,elevation of TNF-α,IL-1β,and IL-6 levels in intestinal mu- cosa,accompanied with IκB-α/NF-κB/ICAM-1 pathway activation. However,breviscapine injection pretreatment could inhibit IκB-α/NF-κB/ICAM-1 pathway activation,leading to reduction of TNF-α,IL-1β,and IL-6 concen-trations in lung,finally attenuate ALI induced by IIR. Conclusion Breviscapine injection pretreatment could atten-uate inflammation in intestine after IIR injury via inhibiting IκB-α/NF-κB/ICAM-1signaling pathway.

Effect of curcumin on NF-κB 、IκB、p-IκB in asthmatic mice / 国际儿科学杂志

Objective To investigate the effect of the asthmatic mice's contents of NF-κB、IκB、p-IκB treated with curcumin.Methods Thirty Balb/c mice were divided into three groups randomly,the normal mice,the asthmatic mice and the curcumin mice.The bronchial hyperresponsiveness of the three groups were exanined by lung function.The NF-κB 、IκB 、p-IκB content of three teams were tested.Results The content of cytoplasm NF-κB in asthmatic mice was lower than the control (P ＜ 0.01).However,the content of cytoplasm NF-κB in curcumin team was more than the asthmatic mice (P ＜ 0.05).In the other hand,The content of nuclear NF-κB in asthmatic mice was more than the control(P ＜0.01).However,the content of nuclear NF-κB in curcumin mice was lower than the asthmatic mice (P ＜ 0.05).The content of cytoplasm p-hκB in asthmatic mice was more than the control and the content of IκB was lower than the control(P ＜ 0.01).Howerer,the content of IκB in curcumin team have much more than the asthmatic mice and the content of p-IκB have much lower than the control(P ＜ 0.01).Condusion Curcumin alleviates the airway hyperresponsiveness of the asthmatic mice through the suppression of NF-κB transcribe to the nuclear via alleviating the phosphorylation of I-κB.

Expression and significance of Maspin and IKKα in sinusoidal mucosa of rats with fungal rhinosinusitis / 中国比较医学杂志

Objective To investigate the expression and significance of Maspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis (FRS).Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group.Another 10 normal rats were used as control group.Mice in the control group were fed with normal diet.In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0.9% NaCl in the abdominal and nasal cavities.In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0.9% NaCl i.p.The mice of the immunosuppressive group were given cyclophosphamide i.p.and 0.9% NaCl injection into the nasal cavity.The invasive FRS group was injected with cyclophosphamide i.p.and Aspergillus fumigatus spore suspension into the nasal cavity.The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining.The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR.Results The serum levels of IL-6 and TNF-α in different groups were significantly different (P 0.05).Theresult of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group (P< 0.05).The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher (P< 0.05).The PCRresult revealed that the expression levels of Maspin and IKKα mRNA were (0.217 ± 0.013) and (0.193 ± 0.012), significantly lower than that in the control, obstruction and immunosuppressive groups [(0.309 ± 0.021), (0.302 ± 0.017), and (0.293 ± 0.02)] (P< 0.05), while the expressions level of IKKα mRNA were significantly higher [(0.319 ± 0.043), (0.384 ± 0.048) vs (0.169 ± 0.015), (0.171 ± 0.018), and (0.175 ± 0.019)] (P< 0.05).Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.

Effects of Biejiajian Pills on NF-κB signaling pathway in hepatic stellate cells in rats / 中成药

AIM To study the effects of Biejiajian Pills (Colla Carapacis Trionycis,Asini Corii Colla,Nidus Vespae,etc.) on NF-κB,p65,p50 and IκB in NF-κB signaling pathway and target gene expression in HSC-T6 cells of rats.METHODS HSC-T6 cells were cultured with Biejiajian Pills drug serum for 24 hours,the expressions of p65,p50,VEGF and TIMP-1 mRNA were determined by qPCR;the expression of p65 was measured by immunofluorescence;the expressions of IκBα,IκBβ and α-SMA were determined by Western blot.RESULTS The Biejiajian Pills middle-,high-dose and positive control groups showed significantly lower expressions of p65,VEGF and TIMP-1 mRNA as compared with the blank control group and negative control group,the expressions of p50 mRNA among various groups showed no significant differences.But immunofluorescence showed that the expression of p65 in cytoplasm was decreased.Meanwhile,Biejiajian Pills showed significantly higher IκBα protein expression and obvious down-regulation of α-SMA expression in a dose-dependent manner,but had no significant influence on the expression of IκBβ.CONCLUSION Biejiajian Pills' therapeutic effects on hepatic fibrosis may be related to influencing NF-κB signaling pathway and inhibiting the expression of down-stream target gene.

Effect of NF-κB on the pathogenic course of non-alcoholic fatty liver disease / 中南大学学报(医学版)

With the economic growth and better standards of living,the prevalence of non-alcoholic fatty liver disease (NAFLD) is expected to increase dramatically worldwide.NAFLD is a common chronic inflammation disease.NF-κB is a transcription factor that plays crucial roles in inflammation,immunity,cell proliferation and apoptosis.It can facilitate the occurrence and development of NAFLD,and the underlying mechanisms are related to insulin resistance,oxidative stress,alteration of intestinal flora,activation ofrenin angiotensin system,etc.

Breviscapine reduces intestinal ischemia reperfusion via inhibiting IκB-α/NF-κB/ICAM-1 pathway in a rat model / 实用医学杂志

Objective To investigate the effects of breviscapine injection on intestinal injury induced by intestine ischemia-reperfusion(IIR). Methods 48 males SD rats with 8-week old were randomly divided into 4 groups:Sham,intestine ischemia-reperfusion(IIR),EB+IIR,TP+IIR. Breviscapine injection 20 mg/(kg·d) was given intraperitoneally in EB + IIR group. TPCA-1(12 mg/kg)was given intravenously 30 min before surgery in TP+IIR group. Rats were subjected to superior mesenteric artery occlusion consisting of 45 min of ischemia and 6 h of reperfusion;sham laparotomy served as controls. Intestine pathology was assayed by H&E staining. Con-centrations of TNF-α,IL-1β,and IL-6 in intestinal mucosa were determined by ELISA. The protein expressions of IκB-α,NF-κB ,ICAM-1of intestine tissue were assayed by western blot. Results IIR induced serious intesti-nal injury ,evidenced as poor intestine pathology ,elevation of TNF-α,IL-1β,and IL-6 levels in intestinal mu- cosa,accompanied with IκB-α/NF-κB/ICAM-1 pathway activation. However,breviscapine injection pretreatment could inhibit IκB-α/NF-κB/ICAM-1 pathway activation,leading to reduction of TNF-α,IL-1β,and IL-6 concen-trations in lung,finally attenuate ALI induced by IIR. Conclusion Breviscapine injection pretreatment could atten-uate inflammation in intestine after IIR injury via inhibiting IκB-α/NF-κB/ICAM-1signaling pathway.

Effects of heat shock protein 70 activator SW02 on lipopolysaccharide-induced expression of inducible nitric oxide synthase in macrophages / 中国药理学通报

Aim To observe the effects of heat shock protein 70 ( Hsp70 ) activator SW02 on lipopolysaccha-ride( LPS)-induced expression of inducible nitric oxide synthase ( iNOS ) and LPS-induced production of nitric oxide ( NO ) in macrophages. Methods RAW264. 7 cells were stimulated by LPS, and were divided into DMSO,DMSO+LPS(1 mg·L-1),SW02,and SW02+LPS ( 1 mg · L-1 ) groups. The protein expression was detected by Western blot. NO concentration was measured by Griess kit. The iNOS mRNA was detected by real-time PCR. The NF-κB binding to iNOS promot-ers was measured by chromatin immunoprecipitation ( ChIP ) assays. Results SW02 significantly blocked the protein and mRNA expression of iNOS as well as the production of NO in LPS-stimulated RAW264 . 7 cells(P0. 05 , SW02 +LPS group vs DMSO+LPS group ) and nuclear translocation of NF-κB ( P > 0. 05 , SW02 + LPS group vs DMSO + LPS group) . However,SW02 reduced the NF-κB binding to iNOS promoters inside the cell( P<0. 05,SW02+LPS group vs DMSO+LPS group) . Conclusion These re-sults show that SW02 prevents iNOS expression and NO induction likely through attenuation of the NF-κB bind-ing to iNOS promoters in macrophages.

Aspirin-Triggered Resolvin D1 Inhibits TGF-β1-Induced EndMT through Increasing the Expression of Smad7 and Is Closely Related to Oxidative Stress

The endothelial-mesenchymal transition (EndMT) is known to be involved in the transformation of vascular endothelial cells to mesenchymal cells. EndMT has been confirmed that occur in various pathologic conditions. Transforming growth factor β1 (TGF-β1) is a potent stimulator of the vascular endothelial to mesenchymal transition (EMT). Aspirin-triggered resolvin D1 (ATRvD1) has been known to be involved in the resolution of inflammation, but whether it has effects on TGF-β1-induced EndMT is not yet clear. Therefore, we investigated the effects of AT-RvD1 on the EndMT of human umbilical vein vascular endothelial cells line (HUVECs). Treatment with TGF-β1 reduced the expression of Nrf2 and enhanced the level of F-actin, which is associated with paracellular permeability. The expression of endothelial marker VE-cadherin in HUVEC cells was reduced, and the expression of mesenchymal marker vimentin was enhanced. AT-RvD1 restored the expression of Nrf2 and vimentin and enhanced the expression of VE-cadherin. AT-RvD1 did also affect the migration of HUVEC cells. Inhibitory κB kinase 16 (IKK 16), which is known to inhibit the NF-κB pathway, had an ability to increase the expression of Nrf2 and was associated with the inhibition effect of AT-RvD1 on TGF-β1-induced EndMT, but it had no effect on TGF-β1-induced EndMT alone. Smad7, which is a key regulator of TGF-β/Smads signaling by negative feedback loops, was significantly increased with the treatment of AT-RvD1. These results suggest the possibility that AT-RvD1 suppresses the TGF-β1-induced EndMT through increasing the expression of Smad7 and is closely related to oxidative stress.

Regulatory effect of a Chinese medicine prescription Hu Qi Shan on the telomerase activity in hepatic precancerous lesions in rats / 中国比较医学杂志

Objective To explore the possible mechanism of action of telomerase in hepatic precancerous lesions, and the regulatory effect of a Chinese medicine prescription HU Qi Shan ( HQS) and its principal drug mistletoe alkali on the telomerase activity in rat liver tissues.Methods Rat model of hepatic precancerous lesions was established by Solt-Farber two-step protocol.The model rats were randomly divided into 5 groups, including the model group, high-dose HQS [8 g/(kg· d)] group, low-dose HQS [4 g/(kg· d)] group, and mistletoe alkali[8 mg/(kg· d)] group.γ-Glutamy-transpeptidase (γ-GT) was analyzed by immunohistochemistry.AFP was detected by immunofluorescence technique.The telomerase activity was detected using a quantitative telomerase detection kit.The expression of NF-κB P65 was detected by immunohistochemistry.The cytoplasmic protein IκB-αwas detected by western blotting.Results After treated with HQS and mistletoe alkali, the areas ofγ-GT-positive foci and number of AFP-positive cells in the liver tissus were significantly decreased than those of the model group ( P<0.05 for both) , the telomerase activity was decreased, the number of NF-κB P65-positive cells was also decreased ( P <0.05 ) , whereas the intracytoplasmic expression of IκB-αproteins was significantly increased ( P <0.05 ) .Conclusions HQS and mistletoe alkali can suppress the telomerase activity.Its possible mechanism may be through inhibition of the over-expressed apoptosis-related genes such as NF-κB P65 and increase the expression of IκB-αdecreasing the telomerase activity.