An Experimental Study on the Role of Nuclear Factor-κB in the Signal Conduction of Protein Kinase C Regulating the Proliferation and Apoptosis of T Lymphocytes in Asthma

To explore the role of nuclear factor-κB(NF-κB) in the signal pathway of protein kinase C (PKC) regulating the proliferation and apoptosis of T lymphocytes in asthma. T lymphocytes were isolated from the asthmatic model of guinea pigs and the asthmatic patients. Either the T cells stimulated with PMA alone or those stimulated with PMA together with pyrrolidine dithiocarbamate (PDTC) were incubated for 1 and 24?h. The proliferation of and the presence of NF-κB in the cells incubated for 1?h were observed by MTT and immunohistochemical staining, respectively. And the cells incubated for 24?h were observed for the apoptosis by TUNEL. All the assays were paralleled with controls, and all the data were analyzedstatistically with the software SAS. The percentage of cells of nuclear positive staining of NF-κB and the proliferation of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated with PMA were significantly higher than those of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated without PMA respectively (P<0.01) and those of T lymphocytes from normal control guinea pigs and normal control persons stimulated with PMA respectively (P<0.01), and were significantly reduced by PDTC (P<0.01). The apoptosis index of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated with PMA were significantly lower than those of T lymphocytes from asthmatic guinea pigs and asthmatic patients stimulated without PMA respectively (P<0.01) and those of T lymphocytes from normal control guinea pigs and normal control persons stimulated with PMA respectively (P<0.01), and were significantly induced by PDTC (P<0.01). There were good positive correlation between the percentage of cells of nuclear staining of NF-κB of T lymphocytes and the proliferation of T lymphocytes (r=0.51-0.72, P<0.001), and also good negative correlation between the percentage of cells of nuclear staining of NF-κB and the apoptosis index of T lymphocytes (r=-0.55-0.71, P<0.001, respectively). It concludes that the active PKC of asthmatic T lymphocytes promoting the proliferation and inhibiting the apoptosis of T lymphocytes may be mediated by activating NF-κB, the activation of PKC-NF-κB signal pathway of T lymphocytes NF-κB may play an important role in the pathogenesis of asthma.

Suppressing high mobility group box-1 release alleviates morphine tolerance via the adenosine5'-monophosphate-activated protein kinase/heme oxygenase-1 pathway

Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.

Hepatitis B virus X protein up-regulates tumor necrosis factor-α expression in cultured mesangial cells via ERKs and NF-κB pathways

Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.

Sphingosine kinase 1 dependent protein kinase C-δ activation plays an important role in acute liver failure in mice

AIM: To investigate the role of protein kinase C(PKC)-δ activation in the pathogenesis of acute liver failure(ALF) in a well-characterized mouse model of D-galactosamine(D-Gal N)/lipopolysaccharide(LPS)-induced ALF.METHODS: BALB/c mice were randomly assigned to five groups, and ALF was induced in mice by intraperitoneal injection of D-Ga IN(600 mg/kg) and LPS(10 μg/kg). Kaplan-Meier method was used for survival analysis. Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels at different time points within one week were determined using a multiparameteric analyzer. Serum levels of high-mobility group box 1(HMGB1), tumor necrosis factor(TNF)-α, interleukin(IL)-1β, IL-6, and IL-10 as well as nuclear factor(NF)-κB activity were determined by enzyme-linked immunosorbent assay. Hepatic morphological changes at 36 h after ALF induction were assessed by hematoxylin and eosin staining. Expression of PKC-δ in liver tissue and peripheral blood mononuclear cells(PBMCs) was analyzed by Western blot.RESULTS: The expression and activation of PKC-δ were up-regulated in liver tissue and PBMCs of mice with D-Gal N/LPS-induced ALF. Inhibition of PKC-δ activation with rottlerin significantly increased the survival rates and decreased serum ALT/AST levels at 6, 12 and 24 h compared with the control group(P < 0.001). Rottlerin treatment also significantly decreased serum levels of HMGB1 at 6, 12, and 24 h, TNF-α, IL-6 and IL-1 β at 12 h compared with the control group(P < 0.01). The inflammatory cell infiltration and necrosis in liver tissue were also decreased in the rottlerin treatment group. Furthermore, sphingosine kinase 1(Sph K1) dependent PKC-δ activation played an important role in promoting NF-κB activation and inflammatory cytokine production in ALF.CONCLUSION: Sph K1 dependent PKC-δ activation plays an important role in promoting NF-κB activation and inflammatory response in ALF, and inhibition of PKC-δ activation might be a potential therapeutic strategy for this disease.

山姜素调控PI3K/Akt/NF-κB信号通路对冠心病大鼠心肌损伤的影响

目的:探讨山姜素调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/核转录因子-κB(NF-κB)信号通路对冠心病(CHD)大鼠心肌损伤的影响。方法:选取健康成年SPF级SD雄性大鼠60只,随机分为空白组、模型组、山姜素小剂量组、山姜素中剂量组、山姜素高剂量组各12只,采用Western blot法检测大鼠PI3K/Akt/NF-κB信号通路蛋白表达,采用ELISA法检测大鼠炎症因子指标[肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)、白介素-18(IL-18)]和氧化应激指标[活性氧(ROS)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)],观察并记录大鼠心肌损伤情况。结果:与空白组比较,模型组的心肌缺血和心肌梗死面积更大(P<0.05),与模型组比较,山姜素小、中、高剂量组的心肌缺血和心肌梗死面积均缩小(P<0.05),且高剂量组的心肌缺血和心肌梗死面积小于山姜素中、小剂量组,中剂量组心肌缺血和心肌梗死面积小于山姜素小剂量组(P<0.05)。与空白组比较,模型组的PI3K、Akt、NF-κB更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的PI3K、Akt、NF-κB均升高(P<0.05),且高剂量组的PI3K、Akt、NF-κB高于山姜素中、小剂量组,中剂量组PI3K、Akt、NF-κB高于山姜素小剂量组(P<0.05)。与空白组比较,模型组的TNF-α、IL-1β、IL-18更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的TNF-α、IL-1β、IL-18均降低(P<0.05),且山姜素高剂量组的TNF-α、IL-1β、IL-18低于中、小剂量组,山姜素中剂量组的TNF-α、IL-1β、IL-18低于山姜素小剂量组(P<0.05)。与空白组比较,模型组的ROS、GSH-Px、MDA更高(P<0.05),与模型组比较,山姜素小、中、高剂量组的ROS、GSH-Px、MDA均降低(P<0.05),且山姜素高剂量组ROS、GSH-Px、MDA低于山姜素中、小剂量组,山姜素中剂量组的ROS、GSH-Px、MDA低于山姜素小剂量组(P<0.05)。结论:山姜素可通过调控PI3K/Akt/NF-κB信号通路改善CHD大鼠心肌炎症和氧化应激状况,进而发挥心肌保护作用。

IKBKE、YAP1和TEAD2在结直肠癌中的表达及临床意义

目的探讨核因子κb激酶亚基ε的抑制剂(IKBKE)、Yes相关蛋白1(YAP1)和转录增强结构域转录因子2(TEAD2)在结直肠癌(CRC)组织中的表达及其临床意义。方法收集2016年1月至2017年12月在蚌埠医科大学第一附属医院手术切除的142例CRC组织及对应癌旁组织,采用免疫组化法检测标本中IKBKE、YAP1和TEAD2的表达情况。分析3种蛋白在CRC组织中表达的相关性,分析蛋白阳性率与患者临床病理参数及预后的关系;绘制Kaplan-Meier生存曲线,比较这些蛋白不同表达情况患者的生存差异。采用Cox回归分析影响患者预后的危险因素。结果CRC组织中IKBKE、YAP1和TEAD2的阳性率均显著高于癌旁组织(65.5%比9.9%,73.9%比14.1%,66.9%比8.5%,均P<0.05)。IKBKE的表达与肿瘤的分化程度、浸润深度、淋巴结转移、肿瘤-淋巴结-远处转移(TNM)分期有关,YAP1和TEAD2的表达均与肿瘤的分化程度、浸润深度、淋巴结转移、远处转移及TNM分期有关。Spearman秩相关分析显示CRC组织中IKBKE与YAP1、TEAD2表达均呈正相关(均P<0.01)。Kaplan-Meier生存分析显示IKBKE、YAP1和TEAD2阳性表达组的总生存率降低。Cox回归分析显示IKBKE、YAP1和TEAD2阳性、肿瘤分化程度高、TNM分期高是CRC患者预后的独立危险因素。结论CRC中IKBKE、YAP1和TEAD2阳性表达与肿瘤的分化程度、TNM分期、转移等因素有关,可能成为CRC治疗的潜在靶点;检测这3个蛋白的表达有助于评估预后。

Metformin inhibits nuclear factor-κB activation and inflammatory cytokines expression induced by high glucose via adenosine monophosphate-activated protein kinase activation in rat glomerular mesangial cells in vitro

Background The renoprotective mechanisms of adenosine monophosphate （AMP）-activated protein kinase （AMPK） agonist-metformin have not been stated clearly.We hypothesized that metformin may ameliorate inflammation via AMPK interaction with critical inflammatory cytokines The aim of this study was to observe the effects of metformin on expression of nuclear factor-κB （NF-κB）,monocyte chemoattractant protein-1 （MCP-1）,intercellular adhesion molecule-1 （ICAM-1） and transforming growth factor-beta 1 （TGF-β1） induced by high glucose （HG） in cultured rat glomerular mesangial cells （MCs）.Methods MCs were cultured in the medium with normal concentration glucose （group NG,5.6 mmol/L）,high concentration glucose （group HG,25 mmol/L） and different concentrations of metformin （group M1,M2,M3）.After 48-hour exposure,the supernatants and MCs were collected.The expression of NF-κB,MCP-1,ICAM-1,and TGF-β1 mRNA was analyzed by real time polymerase chain reaction.Westem blotting was used to detect the expression of AMPK,phospho-Thr-172 AMPK （p-AMPK）,NF-κB p65,MCP-1,ICAM-1,and TGF-β1 protein.Results After stimulated by HG,the expression of NF-κB,MCP-1,ICAM-1,TGF-β1 mRNA and protein of MCs in group HG increased significantly compared with group NG （P ＜0.05）.Both genes and protein expression of NF-κB,MCP-1,ICAM-1,TGF-β1 of MCs induced by high glucose were markedly reduced after metformin treatment in a dose-dependent manner （P ＜0.05）.The expression of p-AMPK increased with the rising of metformin concentration,presenting the opposite trend,while the level of total-AMPK protein was unchanged with exposure to HG or metformin.Conlusion Metformin can suppress the expression of NF-κB,MCP-1,ICAM-1 and TGF-β1 of glomerular MCs induced by high glucose via AMPK activation,which may partlv contribute to its reno-protection.

L-and T-type Ca^(2+)channels dichotomously contribute to retinal ganglion cell injury in experimental glaucoma

Retinal ganglion cell apoptotic death is the main pathological characteristic of glaucoma,which is the leading cause of irreversible blindness.Disruption of Ca^(2+)homeostasis plays an important role in glaucoma.Voltage-gated Ca^(2+)channel blockers have been shown to improve vision in patients with glaucoma.However,whether and how voltage-gated Ca^(2+)channels are involved in retinal ganglion cell apoptotic death are largely unknown.In this study,we found that total Ca^(2+)current densities in retinal ganglion cells were reduced in a rat model of chronic ocular hypertension experimental glaucoma,as determined by whole-cell patch-clamp electrophysiological recordings.Further analysis showed that L-type Ca^(2+)currents were downregulated while T-type Ca^(2+)currents were upregulated at the later stage of glaucoma.Western blot assay and immunofluorescence experiments confirmed that expression of the Ca_(V)1.2 subunit of L-type Ca^(2+)channels was reduced and expression of the Ca_(V)3.3 subunit of T-type Ca^(2+)channels was increased in retinas of the chronic ocular hypertension model.Soluble tumor necrosis factor-α,an important inflammatory factor,inhibited the L-type Ca^(2+)current of isolated retinal ganglion cells from control rats and enhanced the T-type Ca^(2+)current.These changes were blocked by the tumor necrosis factor-αinhibitor XPro1595,indicating that both types of Ca^(2+)currents may be mediated by soluble tumor necrosis factor-α.The intracellular mitogen-activated protein kinase/extracellular signal-regulated kinase pathway and nuclear factor kappa-B signaling pathway mediate the effects of tumor necrosis factor-α.TUNEL assays revealed that mibefradil,a T-type calcium channel blocker,reduced the number of apoptotic retinal ganglion cells in the rat model of chronic ocular hypertension.These results suggest that T-type Ca^(2+)channels are involved in disrupted Ca^(2+)homeostasis and apoptosis of retinal ganglion cells in glaucoma,and application of T-type Ca^(2+)channel blockers,especially a specific CaV3.3 blocker,may be a potential strategy for the treatment of glaucoma.

多囊卵巢综合征伴胰岛素抵抗相关信号通路的研究进展

多囊卵巢综合征(PCOS)是一种好发于青春期及育龄期女性的涉及诸多因素的内分泌代谢性疾病,临床主要表现为闭经、体胖、多毛痤疮以及妊娠率显著降低,但目前其病因病机尚不清楚。胰岛素抵抗(IR)与多种慢性非传染性疾病(高血压、糖尿病、心脑血管疾病、PCOS等)相关,目前已知磷脂酰肌醇-3-激酶/蛋白激酶B、肝激酶B1/AMP活化的蛋白激酶、沉默信息调节因子1/叉头框转录因子O1、Toll样因子4/核因子κB信号通路参与了PCOS伴IR(PCOS-IR)的作用机制,因此充分认识PCOS-IR的发病机制在疾病预防中有重要临床意义。

Scutellarein Ameliorated Chondrocyte Inflammation and Osteoarthritis in Rats

Objective:Osteoarthritis(OA)is a degenerative joint disorder characterized by the gradual degradation of joint cartilage and local inflammation.This study aimed to investigate the anti-OA effect of scutellarein(SCU),a single-unit flavonoid compound obtained from Scutellaria barbata D.Don,in rats.Methods:The extracted rat chondrocytes were treated with SCU and IL-1β.The chondrocytes were divided into control group,IL-1βgroup,IL-1β+SCU 50µmol/L group,and IL-1β+SCU 100µmol/L group.Morphology of rat chondrocytes was observed by toluidine blue and safranin O staining.CCK-8 method was used to detect the cytotoxicity of SCU.ELISA,qRT-PCR,Western blotting,immunofluorescence,SAβ-gal staining,flow cytometry,and bioinformatics analysis were applied to evaluate the effect of SCU on rat chondrocytes under IL-1βintervention.Additionally,anterior cruciate ligament transection(ACL-T)was used to establish a rat OA model.Histological changes were detected by safranin O/fast green,hematoxylin-eosin(HE)staining,and immunohistochemistry.Results:SCU protected cartilage and exhibited anti-inflammatory effects via multiple mechanisms.Specifically,it could enhance the synthesis of extracellular matrix in cartilage cells and inhibit its degradation.In addition,SCU partially inhibited the nuclear factor kappa-B/mitogen-activated protein kinase(NF-κB/MAPK)pathway,thereby reducing inflammatory cytokine production in the joint cartilage.Furthermore,SCU significantly reduced IL-1β-induced apoptosis and senescence in rat chondrocytes,further highlighting its potential role in OA treatment.In vivo experiments revealed that SCU(at a dose of 50 mg/kg)administered for 2 months could significantly delay the progression of cartilage damage,which was reflected in a lower Osteoarthritis Research Society International(OARSI)score,and reduced expression of matrix metalloproteinase 13(MMP13)in cartilage.Conclusion:SCU is effective in the therapeutic management of OA and could serve as a potential candidate for future clinical drug therapy for OA.

Breviscapine attenuates acute pancreatitis by inhibiting expression of PKCα and NF-κB in pancreas

AIM:To study the effect of breviscapine (Bre) on activity of protein kinase Cα (PKCα) and nuclear factor (NF)-κB in pancreas,and the mechanism of Bre attenuating acute pancreatitis (AP). METHODS:One hundred and eight rats were randomly divided into acute necrotizing pancreatitis (ANP) group,Bre group (ANP + Bre group) and sham operation (SO) group,36 rats in each group. ANP model was induced by a retrograde injection of 4% sodium deoxycholate into the bilio-pancreatic duct. Fifteen minutes after the ANP model was induced,the rats in Bre group were intraperitoneally injected with Bre (0.4 mg/100 g body weight or 0.1 mL/100 g body weight). Survival time and mortality of rats were calculated. Serum amylase and malondialdehyde levels were measured,volume of ascites was recorded and morphology of pancreas and lung was evaluated at 1,5 and 10 h,after the ANP model was induced,respectively. Expressions of PKCα and subunit p65 of NF-κB in pancreas were detected by immunohistochemistry and Western blotting. RESULTS:The life span of rats was longer and the mortality was lower in Bre group than in ANP group 13.51 ± 5.46 vs 25.36 ± 8.11 (P < 0.05). The amylase and MDA levels as well as the volume of ascites were lower and the pathological changes in pancreas and lung were less in Bre group than ANP group (P < 0.05),indicating that the pancreatitis is less severe in Bre group than ANP group. The activation of PKCα and NF-κB p65 in pancreas was induced rapidly and reached their peak at 1 h or 5 h after ANP,but their activity in Bre group was significantly inhibited. CONCLUSION:Bre exerts its therapeutic effect on AP by inhibiting the activation of PKCα and NF-κB p65 in pancreas.

IL-36 Cytokine Expression and Its Relationship with p38 MAPK and NF-κB Pathways in Psoriasis Vulgaris Skin Lesions

Summary： This study examined the correlation of the expression of interleukin-36 （IL-36）, a novel member of interleukin-1 （IL-1） family, with p38 mitogen-activated protein kinase （p38 MAPK） and nu clear factor-kappa B （NF-kB） pathways in psoriasis vulgaris skin lesions. The expression levels of IL-36a, IL-3613, IL-367, phosphorylated p38 MAPK, and NF-id3p65 were detected in the skin tissues of 38 psoriasis patients and 17 healthy control subjects by real-time quantitative reverse transcription po lymerase chain reaction （qRT-PCR） and Western blotting. The cytokine expression levels were com pared between the psoriasis group and the control group. A correlation analysis between cytokine pro teins was performed in the psoriasis group. Results showed that the expression levels of IL-36a, IL-3613, IL-36y, phosphorylated p38 MAPK and NF-rh3p65 in the psoriasis group were Significantly higher than those in the control group （P〈0.001）. In the psoriasis group, the IL-36 cytokine expression was positively correlated with phosphorylated p38 MAPK and NF-kBp65 expression （P〈0.05）. A significant positive correlation was also found between the phosphorylated p38 MAPK and NF-v,.Bp65 expression （P〈0.01）. It was concluded that the increased IL-36 expression is correlated with p38 MAPK and NF-kB pathways in psoriasis vulgaris skin lesions. All the three factors may be jointly involved in the pathogenesis and local inflammatory response of psoriasis.

Crotalaria ferruginea extract attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting MAPK/NF-κB signaling pathways

Objective:To evaluate the anti-inflammatory activity of Crotalaria ferruginea extract(CFE)and its mechanism.Methods:An intratracheal lipopolysaccharide(LPS)instillationinduced acute lung injury(ALI)model was used to study the antiinflammatory activity of CFE in vivo.The LPS-induced shock model was used to analyze the effect of CFE on survival.LPS-stimulated RAW264.7 cell model was used to investigate the anti-inflammatory activity of CFE in vitro and the effects on mitogen-activated protein kinase(MAPK)or nuclear factor-κB(NF-κB)signaling pathways.Results:CFE administration decreased the number of inflammatory cells,reduced the levels of tumor necrosis factor-α(TNF-α),monocyte chemotactic protein-1(MCP-1),interleukin-6(IL-6),and interferon-γ,and diminished protein content in the bronchoalveolar lavage fluid of mice.CFE also reduced lung wet-to-dry weight ratio,myeloperoxidase,and lung tissue pathological injury.CFE preadministration improved the survival rate of mice challenged with a lethal dose of LPS.CFE reduced LPS-activated RAW264.7 cells to produce nitric oxide,TNF-α,MCP-1,and IL-6.Furthermore,CFE inhibited nuclear translocation and phosphorylation of NF-κB P65,extracellular signal-regulated kinase,c-Jun N-terminal kinases,and P38 MAPKs.Conclusions:CFE exhibits potent anti-inflammatory activity in LPS-induced ALI mice,LPS-shock mice,and RAW264.7 cells,and its mechanism may be associated with the inhibition of NF-κB and MAPK signaling pathways.Crotalaria ferruginea may be a useful therapeutic drug for the treatment of ALI and other respiratory inflammations.

Protective effect of Jiaweibugan decoction against diabetic peripheral neuropathy

Oxygen free radical damage is regarded as a direct or indirect common pathway associated with diabetic neuropathy and is the main cause of complications in peripheral neuropathies. We speculate that Jiaweibugan decoction has a significant effect in treating diabetic peripheral neuropathy through an anti-oxidative stress pathway. In this study, a diabetic rat model was established by intraperitoneal injection of streptozotocin. Rats were treated with Jiaweibugan decoction via intragastric administration. The levels of malondialdehyde and glutathione, which are indirect indexes of oxidative stress, in serum were determined using a colorimetric method. The expression levels of nuclear factor kappa B p65 mRNA and p38 mitogen-activated protein kinase, which are oxidative stress associated factors, in the dorsal root ganglion of spinal $4-6 segments were evaluated by reverse-transcriptase polymerase chain reaction and immunohistochemistry. Results showed that, Jiaweibugan decoction significantly ameliorated motor nerve conduction velocity in diabetic rats, effectively decreased malondialdehyde levels in serum and the expression of nuclear factor kappa B p65 mRNA and p38 mitogen-activated protein kinase mRNA in the dorsa root ganglion, and increased glutathione levels in serum. Therefore, our experimental findings indicate that Jiaweibugan decoction plays an anti-oxidative stress role in the diabetic peripheral neuropathy process, which has a protective effect on peripheral nerve injury.

Albumin resuscitation protects against traumatic/hemorrhagic shock-induced lung apoptosis in rats

Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rats with induced T/HS; the rats were subjected to femur fracture, ischemia for 30 min, and reperfusion for 20 min with Ringer's lactate solution (RS) or 5% (w/v) albumin (ALB), and the left lower lobes of the lungs were resected. Results: Albumin administered during reperfusion markedly attenuated injury of the lung and decreased the concentration of lactic acid and the number of in situ TdT-mediated dUTP nick-end labelling (TUNEL)-positive cells. Moreover, immunohistochemistry performed 24 h after reperfusion revealed increases in the level of nuclear factor κB (NF-κB), and phosphorylated p38 mitogen-activated protein kinase (MAPK) in the albumin-untreated group was down-regulated by albumin treatment when compared with the sham rats. Conclusion: Resuscitation with albumin attenuates tissue injury and inhibits T/HS-induced apoptosis in the lung via the p38 MAPK signal transduction pathway that functions to stimulate the activation of NF-κB.

FSH-PI3K/AKT/NF-κB蛋白在上皮性卵巢癌组织中的表达及作用机制

目的探究卵泡雌激素(FSH)-磷脂酞肌醇3激酶/蛋白激酶B(PI3K/AKT)/核转录因子κB(NF-κB)蛋白对上皮性卵巢癌的影响。方法选取邯郸市中心医院手术切除的上皮性卵巢癌组织(32例)、卵巢交界性肿瘤组织(32例)、卵巢良性肿瘤组织(32例)、正常卵巢组织(32例),对比不同卵巢组织中FSHR、p-AKT、NF-κB蛋白阳性表达率,分析各蛋白之间相关性,并分析FSH对SKOV-3、3AO细胞生长、侵袭促进作用。结果上皮性卵巢癌组织FSHR、p-AKT、NF-κB蛋白阳性表达率:卵巢交界性肿瘤组织>卵巢良性肿瘤组织>正常卵巢组织,差异有统计学意义(P<0.05);上皮性卵巢癌组织中FSHR、p-AKT、NF-κB蛋白之间呈正相关(P<0.05);SKOV-3细胞于FSH浓度40 mIU/ml作用48 h时细胞增殖率最大,不同浓度,比较差异有统计学意义(P<0.05),不同时间点,差异无统计学意义(P>0.05);3AO细胞于FSH浓度40 mIU/mL作用24 h时细胞的增殖率最大,不同浓度、不同时间点,比较差异有统计学意义(P<0.05);SKOV-3细胞中,与对照组相比,FSH组p-AKT蛋白表达增加,LY294002组、FSH/LY294002组p-AKT蛋白表达减少(P<0.05);FSH组NF-κB蛋白表达高于LY294002组(P<0.05);3AO细胞中,LY294002组、FSH/LY294002组p-AKT蛋白表达少于对照组(P<0.05),FSH组p-AKT蛋白表达高于对照组、LY294002组、FSH/LY294002组(P<0.05);FSH组胞核中NF-κB蛋白表达高于对照组、LY294002组、FSH/LY294002组(P<0.05);4组SKOV-3、3AO细胞中AKT1/2蛋白表达无明显差异(P>0.05);对照组、LY294002组、FSH/LY294002组两种细胞穿过小室的细胞数少于FSH组(P<0.05)。结论 FSH可能是经由PI3K/Akt通路上调卵巢癌细胞株SKOV-3、3AO中NF-κB蛋白表达,促进肿瘤细胞增殖、侵袭。

Metformin alleviates LTA-induced inflammatory response through PPARγ/MAPK/NF-κB signaling pathway in bovine mammary epithelial cells

Mastitis is a common inflammatory cow mammary infection;that causes significant economic loss in dairy industry.Given the interesting connection between metformin’s anti-inflammatory function and mastitis model induced by LTA in pbMECs,our objective was to prove that metformin was beneficial in suppressing proinflammatory response induced by LTA through modulation of mitogen-activated protein kinase(MAPK)and nuclear factor kappa B(NF-κB)signaling pathways and activation of peroxisome proliferator-activated receptor-γ(PPARγ)in pbMECs.The proliferation of cells and mRNA expression were measured using EdU assay and quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR).Immunoblotting and immunofluorescence analysis were conducted to evaluate the expression of target proteins in inflammatory and anti-inflammatory responses to metformin and LTA.Finally,pbMECs were allowed to treat with the PPAR antagonist GW9662,and inflammatory markers were detected in the cells.Our results showed that LTA concentration at 100μg/mL significantly stimulated the MAPK14,IL-6 and IL-1βmRNA expressions compared to the control cells(P<0.05)in dose-dependent tests for LTA.Metformin suppressed the phosphorylation expressions of MAPK(ERK1/2,p38,and JNK)in LTA-stimulated pbMECs.Metformin also reduced the protein expression of NF-κB,interleukin-8(IL-8),interleukin-1β(IL-1β)and interleukin-6(IL-6)in pbMECs pretreated with LTA.Metformin administration activated PPARγphosphorylation by up-regulating the expression of PPARγin LTA-stimulated pbMECs.Treatment with GW9662 resulted in increased IL-6 expression,which was reversed by metformin.These findings collectively indicated that metformin act to attenuate LTA-stimulated inflammatory response in pbMECs by suppressing MAPK and NF-κB activation via a mechanism partially dependent on PPARγactivation.These results suggested that metformin could function as an anti-inflammatory drug in the treatment of mastitis.

Rhamnus crenata leaf extracts exhibit anti-inflammatory activity via modulating the Nrf2/HO-1 and NF-κB/MAPK signaling pathways

Objective:To elucidate the potential anti-inflammatory mechanisms of Rhamnus crenata leaf extracts using RAW264.7 cells.Methods:We used 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay to measure cell viability.Nitric oxide(NO)production was measured using Griess reagent.Western blotting and RT-PCR assays were carried out for analyzing the protein and gene expressions of pro-inflammatory mediators,respectively.Moreover,PD98059(ERK1/2 inhibitor),SB203580(p38 inhibitor),SP600125(JNK inhibitor),and BAY11-7082(NF-κB inhibitor)were used to evaluate the anti-inflammatory mechanism of Rhamnus crenata leaf extract.Results:Rhamnus crenata leaf extracts significantly inhibited the production of the pro-inflammatory mediators such as NO,iNOS,COX-2,IL-1β,and TNF-αin lipopolysaccharide(LPS)-stimulated RAW264.7 cells.Rhamnus crenata leaf extracts also suppressed LPS-induced degradation of IκB-αand nuclear accumulation of p65,which resulted in the inhibition of NF-κB activation in RAW264.7 cells.Additionally,the extracts attenuated the phosphorylation of p38,ERK1/2,and JNK in LPS-stimulated RAW264.7 cells.Moreover,HO-1 expression induced by Rhamnus crenata leaf extracts was significantly downregulated by SB230580,PD98059,SP600125 and BAY11-7082.Conclusions:Rhamnus crenata leaf extract may upregulate HO-1 expression through inhibition of p38,ERK1/2,and NF-κB activation,which may contribute to the anti-inflammatory activity of the extracts.Rhamnus crenata leaf extracts may have great potential for the development of anti-inflammatory drugs to treat acute and chronic inflammatory diseases.

Relationship of high-risk HPV infection with MEKK3 and NF-κB expression in cervical intraepithelial neoplasia tissue

Objective:To study the relationship of high-risk human papilloma virus (HPV) infection with mitogen-activated protein kinase/extracellular signal-regulated kinase 3 (MEKK3) and nuclear factorκB (NF-κB) expression in cervical intraepithelial neoplasia tissue.Methods:125 cases of cervical biopsy specimens between May 2013 and March 2016 were collected. The cervical inflammation specimens, cervical intraepithelial neoplasia specimens and cervical cancer specimens were included in inflammation group, CIN group and malignant group respectively. HPV-DNA typing detection kits were used to determine HPV typing, immunohistochemical kits were used to determine MEKK3 and NF-κB protein expression, and fluorescent quantitative PCR kits were used to determine the mRNA expression of MEKK3, NF-κB and downstream molecules.Results: MEKK3 and NF-κB protein expression in high-risk HPV-positive cervical tissue were significantly higher than those in high-risk HPV-negative cervical tissue (P<0.05), and MEKK3, NF-κB, Bcl-2, XIAP, Bmi-1, TGF-β and Vimentin mRNA expression in high-risk type HPV-positive cervical tissue were significantly higher than those in high-risk HPV-negative cervical tissue (P<0.05);Bcl-2, XIAP, Bmi-1, TGF-β and Vimentin mRNA expression in tissue with positive MEKK3 and NF-κB expression were significantly higher than those in tissue with negative MEKK3 and NF-κB expression (P<0.05).Conclusions:High-risk HPV infection will increase the expression of proliferation genes Bcl-2, XIAP and Bmi-1 as well as invasion genes TGF-β and Vimentin in cervical intraepithelial neoplasia tissue through MEKK3/NF-κB pathway.

延胡索总碱贴片安全性和对神经病理性疼痛大鼠镇痛作用研究

目的 探讨延胡索总碱贴片的安全性及其对神经病理性疼痛大鼠的镇痛作用及其机制。方法 (1)安全性实验:采用单次、多次给药的皮肤刺激性实验及皮肤过敏性实验,评价延胡索总碱贴片的安全性。(2)镇痛实验:取48只雄性SD大鼠,随机分为正常组、假手术组、模型组、布洛芬组、双氯芬酸钠贴片组及延胡索总碱贴片高、中、低剂量组,每组6只。除正常组和假手术组外,其余组大鼠采用坐骨神经慢性压迫损伤方法诱导神经病理性疼痛模型,术后第8天开始,布洛芬组灌胃给药,双氯芬酸钠贴片组及延胡索总碱贴片高、中、低剂量组分别给予双氯芬酸钠贴片及延胡索总碱贴片腹部贴敷,其余组腹部贴敷空白贴片,均每天1次,连续14 d。分别于术前及干预后1,3,7,10,14 d测定各组大鼠热缩足反射潜伏期;采用Western blot法检测各组大鼠干预结束后脊髓中Toll样受体4(TLR4)、核因子κB(NF-κB)、p38丝裂原活化蛋白激酶(p38MAPK)、信号传导和转录激活因子3(STAT3)蛋白表达情况。结果 (1)给予延胡索总碱贴片后,皮肤刺激反应评分均小于0.5分,致敏率为0,组织结构与正常皮肤无差异。(2)镇痛实验中,延胡索总碱贴片中、高剂量组干预后3 d和延胡索总碱贴片各剂量组干预后7,10,14 d患侧的热缩足反射潜伏期均明显长于同期模型组(P均<0.05);干预结束后,延胡索总碱贴片各剂量组大鼠脊髓中TLR4、NF-κB、p38MAPK蛋白相对表达量均明显低于模型组(P均<0.05), STAT3蛋白相对表达量均明显高于模型组(P均<0.05)。结论 延胡索总碱贴片安全性良好,可抑制慢性坐骨神经挤压损伤神经病理性疼痛大鼠痛觉敏感,其机制可能与抑制脊髓中TLR4、NF-κB、p38MAPK表达并上调STAT3的表达有关。