Tongxie Yaofang Regulates T Lymphocyte Subsets to Improve Immune Microenvironment of Colorectal Cancer Under Chronic Stress / 中国实验方剂学杂志

ObjectiveTo explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism. MethodA total of 40 male SPF BABL/C mice were randomized into normal group， stress group， Tongxie Yaofang group （13.65 g·kg-1）， and Tongxie Yaofang-stress group （13.65 g·kg-1）， with 10 in each group. Chronic restraint stress was induced in mice and administration （ig） of Tongxie Yaofang began after 7 days of stress. On the 14th day， forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice. The effect of this prescription on the body mass and tumor volume of mice was observed. After the last administration， mouse serum and tumor samples were collected. The content of T lymphocytes （CD3+， CD4+， CD8+， and CD4+/CD8+） in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone （CORT） in peripheral blood， and interleukin （IL）-2， interferon-γ （IFN-γ）， IL-6， and IL-10 in the serum were determined by enzyme-linked immunosorbent assay （ELISA）. The protein expression of inhibitor of nuclear factor-κB（IκB） kinase α/β （IKKα/β）， nuclear factor-κB （NF-κB）α （IκBα）， NF-κB p65， and phosphorylated （p）-NF-κB p65 was measured by Western blot. ResultCompared with the normal group， the stress group had large tumor volume （P<0.05）， low content of CD3+， CD4+， and CD4+/CD8+ （P<0.05， P<0.01）， high content of CD8+， low content of T helper 1 （Th1）-secreted IFN-γ （P<0.05）， high content of T helper 2 （Th2）-secreted IL-10 （P<0.05） and CORT （P<0.05）， high protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and low protein expression of IκBα （P<0.05）. Compared with the normal group， the Tongxie Yaofang group showed slow tumor growth， high content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， low content of CD8+ （P<0.05）， high content of Th1-secreted IL-2 and IFN-γ （P<0.05）， low content of Th2-secreted IL-6 and IL-10 （P<0.05）， low content of CORT， low protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and high protein expression of IκBα （P<0.01）. Tongxie Yaofang-stress group demonstrated slower tumor growth， higher content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， smaller content of CD8+ （P<0.05）， higher content of IL-2 and IFN-γ （P<0.05）， lower content of IL-6， IL-10 （P<0.05）， and CORT （P<0.05）， lower protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05，P<0.01）， and higher protein expression of IκBα （P<0.01） than the stress group. ConclusionTongxie Yaofang can delay the growth of colorectal cancer under chronic stress and alleviate the deterioration of the immune microenvironment， possibly by inhibiting NF-κB signaling pathway， regulating the function of T lymphocyte subsets， and thus suppressing the secretion of pro-inflammatory factors.

Dihydromyricetin improves cardiac insufficiency by inhibiting HMGB1 in diabetic rats / 南方医科大学学报

OBJECTIVE@#To investigate the effect of dihydromyricetin (DHM) on cardiac insufficiency in diabetic rats and explore the underlying mechanism.@*METHOD@#Twenty-four male SD rats were randomized equally into normal control group, type 2 diabetes (T2DM) group fed on a high-glucose and high-fat diet for 6 weeks with low-dose streptozotocin (STZ) injection, metformin (MET) group with daily intragastric administration of MET (150 mg/kg) for 8 weeks after T2DM modeling, and dihydromyricetin (DHM) group with daily intragastric administration of DHM (250 mg/kg) for 8 weeks after modeling. The levels of fasting blood glucose, low density lipoprotein (LDL-C), triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL-C) and glycosylated hemoglobin (HbA1c) of the rats were measured, and plasma levels of insulin and high mobility group protein-1 (HMGB1) were detected with ELISA. The cardiac function of the rats was assessed using color echocardiography, ECG was measured using a biological signal acquisition system, and myocardial pathology was observed with HE staining. The protein expressions of HMGB1, nuclear factor-κB (NF-κB) p65 and phospho-NF-κB p65 (p-NF-κB p65) in the myocardial tissue were detected using Western blotting.@*RESULTS@#Compared with the control group, the rats in T2DM group showed significant anomalies in cardiac function after modeling with significantly increased plasma HMGB1 level and expressions of HMGB1, NF-κB p65 and p-NF-κB p65 proteins in the myocardial tissue (P < 0.05 or 0.01). Treatment with DHM significantly improved the indexes of cardiac function of the diabetic rats (P < 0.05 or 0.01), decreased plasma HMGB1 level and down-regulated the protein expressions of HMGB1 and p-NF-κB p65 in the myocardial tissue (P < 0.05 or 0.01).@*CONCLUSION@#DHM treatment can improve cardiac function in diabetic rats possibly by down-regulation of HMGB1 and phospho-NF-κB p65 expressions in the myocardium.

Efficacy of Fufang Huangbaiye Tuji on Inflammation on Skin with Deep Ⅱ Degree Burn and Its Dose-effect Relationship / 中国实验方剂学杂志

Objective:To observe the effect of different doses of Fufang Huangbaiye Tuji asin the treatment onof the inflammatory response in healing process for of skin with deep Ⅱ degree burn. Methods in healing process. Methods:The 120 cses patients with deep Ⅱ degree burn of fire-toxin injuring fluid syndrome diagnosed in the affiliated hospital of Chengde Medical University between June 2019 and March 2020 were randomly divided into control group，low -dose treatment group and high -dose treatment group，with 40 cases in each group and once. They got a dressing change perevery day. Control group was locally administered with lodophor solution 35 mL per 1% on the body surface area. Low-dose treatment group was locally administered with compound cortex phellodendri fluid 17.5 mL per 1% on the body surface area，while high-dose treatment group was locally administered with compound cortex phellodendri fluid 35 mL per 1% on the body surface area. Observe theThe inflammatory reaction of wound surface in each group onwas observed at admission and after treatment. The pathological changes of each groupsgroup were observed， and determination of nuclear factor kappa-B（NF-<italic>κ</italic>B） p65 expression inon the wound surface was determined by immunohistochemistry on the 4th day after the treatment. The levels of interleukin（IL）-2，IL-8 and tumor necrosis factor（TNF）-α in wound tissue were measured with ELISA and Bacterial culture and count were performed in each group on the 4^th，10^th and 21^st daydays after treatment. The levels of IL-2，IL-8 and TNF-α in wound tissue were measured with ELISA. Results:There was no significant difference in the degree of wound inflammation in each group at admission，and the degree of relief after treatment was positively correlated with the treatment time. At the simultaneous phase point，the inflammatory reaction was severest in control group，which was followed by low-dose treatment group and high-dose treatment group. Bacterial growth were observed on the 4^th day in control group，which was found in low-dose and high-dose treatment groups on the 10^th day，the detection rates of Staphylococcus aureus and Pseudomonas aeruginosa were the highest. Compared with control group，the mean integrated optical density of NF-<italic>κ</italic>B p65 in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4th day after treatment（<italic>P</italic><0.05），the bacterial count decreased significantly in low-dose and high-dose treatment groups on the 10^th and 21^st days after treatment（<italic>P</italic><0.05），and the levels of IL-2，IL-8 and TNF-<italic>α</italic> in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4^th，10^th and 21^st days after treatment（<italic>P</italic><0.05），with statistically significant differences between low-dose and high-dose treatment groups（<italic>P</italic><0.05）. Histopathological examination showed that inflammatory granulocytes and edema were improved in low-dose and high-dose treatment groups compared with control group，with a more significant performance in high-dose treatment group. Conclusion:The external application of compound cortex phellodendri fluid can reduce thebacterial growth of bacteria in on the wound surface，which may reduce the inflammatory reaction by inhibiting the production and release of inflammatory mediators，with a certain dose-effcteffect relationship，and is worth clinical promotion.

Effect of Betulic Acid on RAGE/NF-κB/iNOS Signaling Pathway Expression in Steatosis Cell / 中国实验方剂学杂志

Objective::To investigate the effect of betulic acid(BA) on steatosis LO2 cells. Method::LO2 cells were intervened with BA at different gradient concentrations (0, 10, 20, 40, 80, 160, 250 μmol·L-1) for 24 hours. methyl thiazolyl tetrazolium(MTT) staining was used to observed cell viability to determine the final concentration of BA. The cells were divided into control, model, dimethylsulfoxide (DMSO) and BA groups, as well as BA groups intervened with low, middle and high concentrations. First, model, DMSO and BA group's cells were cultured in 10% Lipid Mix 1 medium for 24 hours to establish a nonalcoholic fatty liver model. Then, DMSO group and low, medium and high-concentration groups were separately cultured with 0.1%DMSO medium and 20, 40, 80 μmol·L-1 BA medium for 24 hours. And control and model groups were cultured in drug-free medium for 24 hours. Oil red O staining and Nile red staining were used to observe the intracellular lipid droplets. Immunofluorescence was used to detect the protein expression of inducible nitric oxide synthase (iNOS). Western blot was used to detect the protein expression levels of receptor for advanced glycation end-products (RAGE), nuclear factor κB p65 (NF-κB p55) and iNOS. Result::BA within the concentration of 80 μmol·L-1 had no significant toxicity on LO2 cells. Compared with control group, the intracellular lipid droplets were significantly increased in the model group, and the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS also increased significantly(P<0.05). Compared with model group, the intracellular lipid droplets in DMSO group were similar to those in model group, with no significant difference in the three protein expressions between the two groups. However, the intracellular lipid droplets deposition in the BA group was significantly decreased. And the expressions of RAGE, NF-κB p65 and iNOS proteins in high-concentration BA group were significantly decreased(P<0.05, P<0.01). Conclusion::BA can significantly improve the intracellular fat deposition in LO2 cells, which was probably related to the inhibition of the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS.

Protective Effect of Liuwei Dihuangwan on Diabetes Mellitus with Liver Injury Based on SIRT6/NF-κB Signaling Pathway / 中国实验方剂学杂志

Objective: To investigate the protective mechanism of Liuwei Dihuangwan on diabetes mellitus with liver injury in terms of inflammation. Method: The 18 db/db mice were selected from animal model of spontaneous type 2 diabetes, mice were randomly divided into model group, Liuwei Dihuangwan group(9.75 g·kg-1·d-1,once a day), resveratrol group(42.6 mg·kg-1·d-1,once a day)based on fasting blood-glucose (FBG). The 12 litter wild db/m mice were randomly divided into normal group, Liuwei Dihuangwan group(9.75 g·kg-1·d-1,once a day). The normal group and the model group were given the same amount of distilled water by gavage. The mice in each group were treated for 16 weeks. FBG, triglyceride (TG) and alanine aminotransferase (ALT) were examined. Histopathological changes were observed by liver biopsy hematoxylin-eosin (HE) staining. Silent information regulator 6 (SIRT6),nuclear factor-kappaB p65 (NF-κB p65),monocyte chemoattractant protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in the liver tissue were detected by Western blot. Result: Compared with normal group, FBG, TG and ALT in model group increased significantly(PκB p65,MCP-1,VCAM-1 and ICAM-1 in model group were significantly up-regulated(PPPPκB p65, MCP-1 and VCAM-1 were significantly decreased (PPConclusion: Liuwei Dihuangwan can protect the diabetes mice from liver injury, and its mechanism is related to the improvement of lipid metabolism and the inhibition of inflammatory response.

Effects of diammonium glycyrrhizinate on Toll-like receptor 4 in alveolar epithelial cells induced by paraquat / 中华急诊医学杂志

Objective To investigate the effects of diammonium glycyrrhizinate (DG) on Toll-like receptor 4 in rat alveolar epithelial cells induced by paraquat (PQ).Methods Rats in the PR and PDR groups were induced 2 h by RU486 (100 nmol/L).Then rats in the DG and PDR groups were induced 2 h by DG (0.6 mg/mL).Finally,PQ (0.6 mg/mL) were administered and induced 24h in the PQ,DG,PR and PDR groups,while,the NE group was induced 24 h by absolute ethyl alcohol (0.33 μ mol/L),and the NS group was induced 24 h without drugs.MTT assay was used to measure the cell growth and inhibitioneffects of PQ and DG on cells.The ELISA assay was applied to measure the levels of the TLR-4,Myd88,NF-κB P65 and GR.The gene expressions ofTLR-4,Myd88,NF-κB P65 and GR were detected by RT-PCR.Results The survival rate of rat alveolar epithelial cells was decreased by PQ (200,400,600,800,1 000,1 500,2 000 μmol/L),and the IC50 value for 24 h was 927.045 μmol/L.The inhibition rates were (11.74±1.44)％,(18.76±1.30)％,(28.74±0.54)％,(40.30±0.55)％,(51.24±0.76)％,(68.19±1.10)％,(83.16±0.59)％ in the 200,400,600,800,1 000,1 500,2 000 pmol/L PQ treatment groups,respectively.And the inhibition rates were (48.01±1.37)％,(40.68±2.33)％,(32.76±4.11)％,(34.12±4.3)％,(39.22±2.23)％,(51.26±-0.39)％ in the 0.2,0.4,0.6,0.8,1,and 2 mg/mL DG treatment groups,respectively.The levels of TLR4,Myd88,NF-κB P65,and TNF-a in the PQ and PR groups were higher than those in the NS group (all P＜0.01).While,the levels of GR in the PQ and PR groups were lower than that in the NS group (all P＜0.01).And,the levels of TLR4,Myd88,NF-rκB P65,and TNF-α in the DG and PDR groups were lower than those in the PQ group (all P＜0.01).But the levels of GR in the DG and PDR groups were higher than that in the PQ group (all P＜0.01).Conclusions Diammonium Glycyrrhizinate can attenuate the injury of rat alveolar epithelial cells induced by paraquat,can decrease the levels of TLR-4,Myd88,NF-KB and TNF-α,and increase the GR level.

Protective effect of hydroxysafflor yellow A on bleomycin- induced pulmonary inflammation and fibrosis in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of hydroxysafflor yellow A (HSYA), an active ingredient of a traditional Chinese herbal medicine Carthamus tinctorius L., on lung inflflammation and pulmonary fibrosis induced by bleomycin (BLM) in rats.</p><p><b>METHODS</b>Animals were divided into 6 groups including normal group, model group, three HSYA groups and dexamethasone (DXM) group. Three doses of HSYA (35.6, 53.3, and 80.0 mg•kg•day) were intraperitoneally (i.p.) injected in rats for 3 weeks after BLM administration and DXM was used as the positive control (n=8 or 12). Arterial blood gas was assayed and morphological changes were observed. Lung mRNA expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and some cytokines in lung tissue were detected by real-time polymerase chain reaction. Nuclear factor-κB p65 or α-smooth muscle actin (α-SMA) protein distribution in rat lung tissue was observed by immunohistochemistry.</p><p><b>RESULTS</b>On the 7th day after BLM administration, lung tissue showed serious inflammation. Treatment with HSYA or DXM ameliorated lung inflammation. After treatment with HSYA or DXM, oxygen partial pressure (PaO) increased (HSYA 80.0 mg•kg, P<0.01) and COpartial pressure (PaCO) decreased (HSYA 53.3, 80.0 mg•kg, P<0.05). Moreover, the mRNA expression of TNF-α, IL-1β, and IL-6; and the number of NF-κB p65 positive cells was lower in HSYA 53.3 and 80.0 mg•kggroups than those in the model group (all P<0.05). Twenty-one days after BLM administration, HSYA or DXM treatment ameliorated fibrosis, increased PaO2 (HSYA 53.3, 80.0 mg•kg-1, P<0.01), and decreased PaCO2 (53.3 and 80.0 mg•kg-1, P<0.05). Further, the mRNA expression of TGF-β1, α-SMA, and collagen I as well as the number of α-SMA positive cells increased in the model group and HSYA can attenuate these changes (53.3, 80.0 mg•kg, P<0.05). Hematoxylin and eosin and Masson's trichrome staining indicated that the fibrosis and collagen deposition were ameliorated in HSYA groups (53.3, 80.0 mg•kg, P<0.05).</p><p><b>CONCLUSION</b>HSYA could alleviate acute lung inflflammation and chronic pulmonary fibrosis induced by BLM in rats.</p>

Effects of electroacupuncture on inflammatory response of cardiac muscle tissue in mice with acute myocardial ischemia / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) on inflammatory reaction of acute myocardial ischemia (MI) in mice, and to explore its action mechanism.</p><p><b>METHODS</b>Forty adult male C57BL/6 mice were randomly divided into a control group, a sham operation group, a model group and an EA group, 10 mice in each one. The model was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the EA group were treated with EA at "Neiguan" (PC 6) with 2 mA of intensity and 2 Hz /100 Hz of frequency; EA was given 30 min per treatment, once a day for totally 5 days. The mice in the control group and model group were treated with immobilization and no EA was given. The mice in the sham operation group were not treated with ligating at the left anterior descending branch of coronary artery, but the remaining procedure was identical to the model group. The electrocardiogram was recorded and △ST was calculated to evaluate the model. TTC and HE staining methods were applied to evaluate the infarct size and pathologic change of myocardial tissue, respectively. Western blot method was applied to test the protein expression levels of tumor necrosis factor-α (TNF-α), nuclear factor-κB p65 (NF-κB p65), interleukin-1β (IL-1β) and interleukin-8 (IL-8).</p><p><b>RESULTS</b>Compared with the sham operation group, the S-T segments in the model group and EA group were increased obviously after modeling (both <0.01), indicating the MI model was established successfully. The TTC and HE staining results indicated, compared with the sham operation group, the model group had larger infarction size (<0.01), more myocardial fibers injury and inflammatory infiltration; compared with the model group, the infarction size of the EA group was significantly reduced (<0.01), and the myocardial fibers injury and inflammatory infiltration were improved. Compared with the control group, the protein expression levels in the sham operation group were similar (all >0.05); compared with the sham operation group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly increased in the model group (<0.01, <0.05); compared with the model group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly reduced in the EA group (all <0.05).</p><p><b>CONCLUSION</b>EA might reduce the protein expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 in cardiac muscle tissue to inhibit inflammatory reaction and achieve myocardial protective effect in mice with acute myocardial ischemia.</p>

Pinocembrin alleviates hypoxia reoxygenation injury in rat liver cell line BRL-3A / 基础医学与临床

Objcetive To investigate protective effect of pinocembrin ( PIN)on hepatocytes induced by hypoxia/reoxygenation ( H/R) as well as its relationship to the TLR 4/NF-κB signaling pathway .Methods The cells were randomly divided into 4 groups: control group, PIN group, hypoxia/reoxygenation injury group and PIN pretreatment group .The cell viability was measured with CCK-8.The apoptosis rate was determined by Annexin V-FITC/PI staining.The activity of ALT was detected .ELISA was used to evaluate the contents of TNF-αand IL-β.The mRNA and protein expression level of TLR 4, IκB-αand NF-κB P65 in cells was observed by quantita-tive real-time PCR or Western blot .Results The H/R stimulation decreased cell survival rate and enhanced the apoptosis .The activity of ALT was increased .The contents of TNF-αand IL-1βwere significantly enhanced , and the expression level of TLR4 and NF-κB P65 was markedly increased while IκB-αdecreased.After pretreatment with PIN, the cell survival rate increased while the apoptosis rate decreased .The activity of ALT was decreased. TNF-αand IL-1βwere reduced significantly and the expression level of TLR 4 and NF-κB P65 were decreased while IκB-αincreased.Conclusions PIN has protective effects on hypoxia/reoxygenation injury, which might be mediated in part by TLR4/NF-κB signaling pathway .

TLR4/NF-κB signal pathway mediate HBD-2 in patients with preterm premature rupture of membrane associated with histologic chorioamnionitis / 中国综合临床

Objective To observe the effect of human β-defensins-2(HBD-2) for chorioamnionitis(HCA) pregnant women before term premature rupture of fetal membrane(PROM) process,and explore toll-like receptor 4 / nuclear factor-κ B (TLR4 / NF-κB) predominate role in the process of signal transduction pathway in the mechanism.Methods Fifty five women with PROM were enrolled in the study.According to the Results of pathological diagnosis of membranes,pregnant women with PROM divided into histological chorioamnionitis,HCA and non-HCA.The same sample without PROM pregnancies matching the same gestational ages were recruited as control group.We examined the messenger RNA(mRNA) of TLR4,NF-κB p65 and HBD-2 in placenta and fetal membrane real-time reverse transcription polymerase chain reactions by dsDNA-binding dyes of SYBR Green.Results (1)In the placenta,the level of TLR-4(17.15±4.52),NF-κB p65(47.11±14.23),HBD-2mRNA(27.35±2.67) in PROM group were significantly higher than the level of TLR-4(7.21±3.25),NF-κB p65(30.51±13.05),HBD-2mRNA(13.55±0.8) in control group(t=-1.966,-1.474,-1.754,P0.05).Conclusion Linear positive correlation of TLR4,NF-κB and HBD2 indicated that TLR4/NF-κB/HBD2 signal transduction pathway may be involved in the development of preterm premature rupture of membrane associated with histologic chorioamnionitis.

Effects of Shenqi Bufei Tang Decoction on the Expression of HDAC2 and NF-κB p65 in Airway Smooth Muscle Tissues of COPD Rats with Lung-Qi Deficiency Syndrome / 医药导报

Objective To examine the effects of Shenqi Bufei Tang decoction on the expression of histone deacetylase-2 ( HDAC2) and nuclear factor-κB p65 ( NF-κB p65) in the airway smooth muscle tissues of COPD rats with lung-qi deficiency syndrome. Methods A total of 40 male rats were randomly divided into normal control group,model control group,Shenqi Bufei Tang decoction group,and aminophylline group.The COPD rat model with lung-qi deficiency syndrome was established by intra-tracheal instillation of lipopolysaccharide (LPS) and passive smoking for 28 days.Pathological changes of lung tissues were ob-served under the light microscope and the thickness of the small airway wall and airway smooth muscle ( ASM) layer analyzed by the image analysis.Immunohistochemistry,real-time PCR and Western blotting were used to detect the expressions of NF-κB p65 and HDAC2 in ASM. Results The thickness of the airway wall and ASM,and the expression levels of NF-κB p65 mRNA and protein were significantly increased in the model control group when compared with those in the normal control group ( P0.05). Conclu-sion Shenqi Bufei Tang decoction can inhibit the proliferation of ASM in COPD rats with lung-qi deficiency syndrome,which may be associated with the increased expression of HDAC2 and decreased expression of NF-κB p65.

Influence of ginsenoside Re on vascular intima hyperplasia and NF-κB p65 signaling pathway in balloon-injured rats / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the inhibitory effect of ginsenoside Re on intimal hyperplasia induced by bal-loon-injury and to explore the role of NF-κB p65 signaling pathway in the process.METHODS:SD rats (n=40) were di-vided into 5 groups randomly: sham operation group, model group, low-dose ginsenoside Re group, middle-dose ginsen-oside Re group and high-dose ginsenoside Re group.The carotid artery intima injury model was established by 2F balloon catheters in all groups except the sham operation group.The day after modeling, the animals in model group and sham op-eration group were administered intragastrically with distilled water, and the rats in low-dose, middle-dose and high-dose ginsenoside Re groups were given ginsenoside Re at doses of 12.5 mg/kg, 25mg/kg and 50 mg/kg, respectively.After 14 continuous days, the morphological changes of the injured arteries were observed by HE staining and the lumen area, intima area and media area as well as the ratio of intimal area/media area were determined.The expression of tumor necrosis fac-tor-α( TNF-α) and interleukin-1β( IL-1β) were detected by real-time PCR.The proliferating cell nuclear antigen ( PC-NA) and nuclear factor-kappa B ( NF-κB) p65 were examined by immunohistochemistry.RESULTS:Compared with sham operation group, the vessel cavity was narrowed (P sion of PCNA and NF-κB p65 were decreased in medium and high-dose ginsenoside Re groups (P<0.05).CONCLU-SION:Ginsenoside Re inhibits the vascular neointimal hyperplasia induced by balloon-injury in rats, and the molecular mechanism may be related to the inhibition of NF-κB p65 signaling pathway.

Vaccinium bracteatum Thunb. Exerts Anti-Inflammatory Activity by Inhibiting NF-κB Activation in BV-2 Microglial Cells

This study was designed to evaluate the pharmacological effects of Vaccinium bracteatum Thunb. methanol extract (VBME) on microglial activation and to identify the underlying mechanisms of action of these effects. The anti-inflammatory properties of VBME were studied using lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. We measured the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin E₂ (PGE₂), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) as inflammatory parameters. We also examined the effect of VBME on intracellular reactive oxygen species (ROS) production and the activity of nuclear factor-kappa B p65 (NF-κB p65). VBME significantly inhibited LPS-induced production of NO and PGE₂ and LPS-mediated upregulation of iNOS and COX-2 expression in a dose-dependent manner; importantly, VBME was not cytotoxic. VBME also significantly reduced the generation of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6. In addition, VBME significantly dampened intracellular ROS production and suppressed NF-κB p65 translocation by blocking IκB-α phosphorylation and degradation in LPS-stimulated BV2 cells. Our findings indicate that VBME inhibits the production of inflammatory mediators in BV-2 microglial cells by suppressing NF-κB signaling. Thus, VBME may be useful in the treatment of neurodegenerative diseases due to its ability to inhibit inflammatory mediator production in activated BV-2 microglial cells.

A study on Qingshen granule for treatment of renal fibrosis in patients with chronic renal failure accompanied by damp-heat syndrome and its mechanism / 中国中西医结合急救杂志

Objective To observe the anti-fibrosis therapeutic effect and mechanism of Qingshen granule for treatment of patients with chronic renal failure (CRF) accompanied by damp-heat syndrome.Methods Sixty-eight patients with CRF accompanied by damp-heat syndrome were randomly divided into a control group and a observation group, and the study was completed only in 61 patients, 31 in the control group and 30 in the observation group. Thirty subjects having taken physical health examination were assigned in a healthy control group. All the patients in both treatment groups were treated with conventional western medical therapy and traditional Chinese medicine (TCM) retention enema, and for patients in observation group, Qingshen granule was given additionally, 1 bag (10 g) thrice a day taken orally. The therapeutic course was 8 weeks. The clinical therapeutic effect, the levels of serum creatinine (SCr), the glomerular filtration rate (eGFR), serum interleukin-17 (IL-17), collagen type Ⅲ (Col-Ⅲ) and nuclear factor-κB p65 (NF-κB p65) in peripheral blood mononuclear cells (PBMC) were measured before and after treatment in the two treatment groups, and the above results were compared with those in healthy control group.Results Clinically, the total effective rates of the disease and of the TCM syndrome in observation group were significantly higher than those in the control group (86.67% vs. 58.06%, 83.33% vs. 45.16%, bothP < 0.01). In the observation group, the level of SCr was obviously lower, and the level of eGFR was markedly higher after treatment, and compared with the control group, the changes in above data after treatment in observation group were more significant [SCr (μmol/L): 250.62±164.97 vs. 393.72±183.64, eGFR (mL·min-1·1.73 m-2): 33.42±17.24 vs. 39.72±23.85, bothP < 0.05]. After treatment, the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC were obviously lowered in both treatment groups compared with those before treatment, the therapeutic effect in observation group being superior to that in the control group [IL-17 (ng/L): 17.47±8.87 vs. 25.51±16.69, Col-Ⅲ (μg/L): 17.06±8.76 vs. 23.77±10.44, NF-κB p65 (μg/L): 0.58±0.34 vs. 0.83±0.30, allP < 0.05].Conclusion The Qingshen granule can ameliorate the clinical symptoms, improve renal function, decrease the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC, intervene renal fibrosis in patients with CRF and damp-heat syndrome, ultimately delaying the progress of CRF.

The mechanisms of protective effects of erythropoietin (EPO) on sepsis induced myocardial injury in rat / 中华急诊医学杂志

Objective To investigate the protective effects of erythropoietin (EPO) on myocardium injury after sepsis in rats in order to clarify the mechanisms.Methods The rat models of sepsis were produced by cecal ligation and perforation method (CLP).Ninty-six healthy SD rats were randomly (random number) divided into 3 groups:the sham operation group (Sham group,n =32),the sepsis group (CLP group,n =32),and the EPO treatment group (EPO group,n =32) treated with EPO 1000 IU/kg intraperitoneal injection after the CLP.The observation intervals were set at 3 h,6 h,12 h and 24 h after the surgery.The cardiac hemodynamics of the CLP group were measured.Plasma levels of inflammatory factors and myocardial enzyme indicators were determined by enzyme-linked immunosorbent assay (ELISA) ; Membrane potential levels of chondriosome of myocardial cells,cell apoptosis rates and expressions of nuclear factor-κB p65 of myocardium tissue were detected by flow cytometer; Then the pathological change of myocardium with HE staining was observed under light microscopy.Results (1) Compared with the CLP group,left ventricle systolic pressure (LVSP),left ventricle diastolic end pressure (LVEDP),the maximum rate of left ventricle rise and fall (+ dp/dtmax and-dp/dtmax) in the EPO group improved (P ＜0.05,P＜0.01); (2) Compared with the Sham operation group,plasma levels of tumor necrosis factoralpha (TNF-α),interleukin-6 (IL-6),C-reactive protein (CRP),cardiac troponin-I (cTNI),creatine kinase (CK),lactate dehydrogenase (LDH) and glutamine-oxaloacetic transaminase (GOT) in the CLP group increased at each interval (P ＜ 0.05),and those biomarkers in the EPO group were lower than those in the CLP group (P ＜ 0.05) ; On the contrary,plasma level of anti-inflammatory factor IL-10 in EPO group was higher than that in the CLP group (P ＜ 0.01) ; (3) Compared with the Sham operation group,the cell apoptosis rates in the CLP group increased significantly (P ＜ 0.01),and it decreased obviously in the EPO group (P ＜ 0.01); (4) Compared with the Sham group,membrane potential levels of chondriosome of myocardial cell in the CLP group decreased (P ＜0.01),while it increased in the EPO group in comparison with the CLP group (P ＜ 0.01) ; (5) Pathological changes of myocardium after the CLP could be lessened by the EPO treatment.Conclusions EPO may increase membrane potential levels of chondriosome and decrease the apoptosis rates of myocardial cell in rats after sepsis,and it may reduced the production of inflammatory factors to protect the myocardial cell by down-regulating NF-κB p65.Both increased membrane potential level of chonriosome and decreased inflammatory factor may implicate in myocardium protection thereby improving cardiac function after sepsis.

Cardioprotective effect of erythropoietin on sepsis-induced myocardial injury in rats / 世界急诊医学杂志（英文）

BACKGROUND:Sepsis-induced myocardial injury is one of the major predictors of morbidity and mortality of sepsis. The cytoprotective function of erythropoietin (EPO) has been discovered and extensively studied. However, the cardioprotective effects of EPO on sepsis-induced myocardial injury in the rat sepsis model has not been reported.METHODS:The rat models of sepsis were produced by cecal ligation and perforation (CLP) surgery. Rats were randomly (random number) assigned to one of three groups (n=8 for each group):sham group, CLP group and EPO group (1000 IU/kg erythropoietin). Arterial blood was withdrawn at 3, 6, 12, and 24 hours after CLP. cTnI, BNP, CK-MB, LDH, AST, TNF-α, IL-6, IL-10, and CRP were tested by the ELISA assay. Changes of hemodynamic parameters were recorded at 3, 6, 12, 24 hours after the surgery. Histological diagnosis was made by hematoxylin and eosin. Flow cytometry was performed to examine cell apoptosis, myocardium mitochondrial inner membrane potential, and NF-κB (p65). Survival rate at 7 days after CLP was recorded.RESULTS:In the CLP group, myocardial enzyme index and inflammatory index increased at 3, 6, 12 and 24 hours after CLP compared with the sham group, and EPO significantly blocked the increase. Compared with the CLP group, EPO significantly improved LVSP, LV +dp/dtmax, LV -dp/dtmin, and decreased LVEDP at different time. EPO blocked the reduction of mitochondrial transmembrane potential, suppressed the cardiomyocyte apoptosis, inhibited the activation of NF-κB, and reduced the production of proinflmmatory cytokines. No difference in the survival rate at 7 days was observed between the CLP group and the EPO group.CONCLUSION:Exogenous EPO has cardioprotective effects on sepsis-induced myocardial injury.

Effects of endotoxin on NF-κB mRNA expression and aldosteron secretion in rat hepatic stellate cells / 中华临床感染病杂志

Objecfive To investigate the effects of endotoxin on nuclear factor-κB p65(NF-κB p65)mRNA expression and ahtosteron secretion in rat hepatic stellate cells(HSCs).Methods Cultured rat HSCs(HSC-T6)were divided into endotoxin-treated group and control group.Cells in endotoxin-treated group were exposure to 1 mg/ml.endotoxin.Aldosteron secretions of HSCs were determined by radioimmunoassay,and NF-κB p65 mRNA expressions of HSCs were detected by one-step RT-PCR.Results At 6,12,24 and 48 h,aldosteron secretions in endotoxin-treated group were significantly hisher than those in the control group(t=3.063,4.577,6.847 and 9.317,P<0.05),and the expressions of NF-κB p65 mRNA in endotoxin-treated group were also higher than those in control group(t=5.155,6.095,7.875 and 9.313,P<0.01).Aldosteron secretions and NF-κB p65 mRNA expressions in HSCs displayed a positive correlation(r=0.886,P<0.01).Conclusion Endotoxin can up-regulate the aldosteron secretion and NF-κB p65 mRNA expression in rat HSCs,which may be one of the mechanisms of liver fibrosis induced by endotoxin.

Oxymatrine Improves TNBS-induced Colitis in Rats by Inhibiting the Expression of NF-κB p65 / 华中科技大学学报（医学）（英德文版）

Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-κB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid(TNBS)-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats.The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model,which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-κB p65in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-κB p65 in colon tissue.