Objective To explore the effectiveness of the transforming growth factor-β1(TGF-β1) and tumor necrosis factor-α(TNF-α) inducing human bronchial epithelial(HBE) cells to optimize epithelia-mesenchymal transformation(EMT) model. Methods Blank control, TGF-β1 10 ng/ml, TNF-α 10 ng/ml, TGF-β1 10 ng/ml+TNF-α 10 ng/ml induced human epithelial cells for 24 hours. Then the change of morphological alteration were observed by applying CCK8, cells migration assay and Western blot technique. Results When TGF-β1 plus TNF-α induced human epithelial cells for 24 hours, most of HBE cells traits changed including morphological alteration from cobblestone to fusiform, connection between cells vanishing, intercellular space broadening. In the experiments of checking cell migration capacity by the vitro scratch test, the group spacing was 420.06±10.38 μm in the blank control group, 499.86±34.00 μm in the TGF-β1 10 ng/ml group, 514.93±10.56 μm in the TNF-α 10 ng/ml group, 569.68±33.58 μm in the TGF-β1 10 ng/ml+TNF-α10 ng/ml group. TGF-β1 cooperated with TNF-α led to scratch spacing narrowing significantly. Western blot analysis showed that expression of E-cadherin and Vimentin varied significantly in the TGF-β1+TNF-α group. Conclusion Inducing human bronchial epithelial cell by TGF-β1 cooperated with TNF-α optimizes EMT model.
Objective To investigate the effect of heme oxygenase 1 (HO-1) on the apoptosis of human degenerated nucleus pulposus (NP) cells induced by tumor necrosis factor α (TNF-α), and explore its possible molecular mechanism. Methods The intervertebral disc tissues were derived from patients with lumbar intervertebral disc herniation. Then, the NP cells were cultured in vitro and the third generation of NP cells were used for subsequent experiments. Cell counting kit 8 (CCK-8) method was used to observe the proliferative effect of TNF-α on the NP cells in vitro at the concentration of 10, 20, 50, 100, and 200 ng/mL. The most apropriate concentration was selected according to the result of CCK-8. The NP cells were cultured with basal medium (control group), TNF-α (TNF-α group), TNF-α and CoPP 10 μmol/L (CoPP group), and TNF-α and ZnPP 15 μmol/L (ZnPP group), respectively. After cultured, the cell poptosis was detected by Hoechst staining and flow cytometry; the expression of cleaved Caspase-3, epithelial membrane protein 1 (EMP-1), HO-1, and p-P65 proteins were detected by Western blot. In order to further explore the potential molecular mechanisms of HO-1 for cell apoptosis, the NP cells were cultured with TNF-α (TNF-α stimulated group), TNF-α and pyrrolidine dithiocarbamate (PDTC) 5 μmol/L (TNF-α+PDTC stimulated group), respectively. Then the cell apoptosis rate was measured by flow cytometry at 24 hours after cultured. Results The optimal concentration of TNF-α was 100 ng/mL. Hoechst staining showed that a few apoptotic cells could be observed in control group and CoPP group; the apoptosis-like nucleis were observed in TNF-α group and ZnPP group, which was the most significant in ZnPP group. Flow cytometry showed that the cell apoptosis rates of TNF-α group, CoPP group, and ZnPP group were significantly increased when compared with the control group (P<0.05). Compared with TNF-α group, the cell apoptosis rate in CoPP group decreased (P<0.05), while in ZnPP group it increased (P<0.05). Western blot showed that the expression of HO-1 protein in TNF-α group was decreased, and the expressions of cleaved Caspase-3, EMP-1, and p-P65 proteins were increased when compared with the control group (P<0.05). Compared with TNF-α group, the expression of HO-1 protein in CoPP group increased, and the expressions of cleaved Caspase-3, EMP-1, and p-P65 proteins were reduced (P<0.05); the expression of HO-1 protein in ZnPP group decreased (P<0.05), the expressions of cleaved Caspase-3 and EMP-1 proteins increased (P<0.05), and the expression of p-P65 protein was not significantly changed (P>0.05). Compared with TNF-α stimulated group, the cell apoptosis rate in TNF-α+PDTC stimulated group was significantly reduced (t=3.076, P=0.031). Conclusion HO-1 can inhibit the apoptosis of degerated NP cells induced by TNF-α, and its mechanism effect is by inhibiting the nuclear factor кB signaling pathway.
ObjectiveTo investigate the efficacy and safety of recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein (rhTNFR:Fc) for treatment of active rheumatoid arthritis (RA). MethodsThis study included 86 patients with active rheumatoid arthritis treated between September 2011 and January 2013. They were divided into two groups randomly. Forty-three patients in the treatment group received rhTNFR:Fc (25 mg, twice a week) by subcutaneous injection and methotrexate (MTX) (10 mg, orally once a week), and the other 43 patients in the contrast group received MTX (10 mg, orally once a week), hydroxychloroquine (100 mg, orally twice daily), and leflunomide (10 mg, orally once daily). The clinical efficacy of the treatments 12 weeks later were compared between the two groups. American College of Rheumatology (ACR) 20, 50, and 70 evaluation criteria were used for efficacy evaluation. ResultsThe ACR 20, 50 and 70 effective rates in 4, 8 and 12 weeks after the treatment in the treatment group were significantly higher than those in the control group (P<0.05). The seven indicators including the duration of morning stiffness, joint tenderness index, joint swelling index, erythrocyte sedimentation rate, C-reactive protein, platelets and rheumatoid factors within 12 weeks after treatment were significantly improved in both the two groups, and the improvements in the treatment group were more significant (P<0.05). There was no significant difference in the incidence of adverse drug reactions between the two groups (P>0.05). ConclusionRhTNFR:Fc is effecive and safe in treating active RA.
摘要:目的: 研究蜕皮甾酮对非酒精性脂肪性肝病大鼠模型肿瘤坏死因子α(TNFα)与核因子κB(NFκB)表达的影响，并探索其可能的作用机制。 方法 :健康成年SD大鼠36只,随机分为正常对照组12只与实验组24只;正常对照组喂以普通基础饲料,实验组应用高脂饲料喂养。实验12周末时将造模成功的实验组大鼠随机分为模型组与蜕皮甾酮治疗组2个亚组,每组12只;正常对照组喂以普通基础饲料至16周,模型组继续应用改良高脂饲料喂养至16周,蜕皮甾酮治疗组大鼠在高脂饮食同时加用蜕皮甾酮灌胃。实验16周末时处死3组所有大鼠;检测肝脏指数,血清与肝组织生化指标及肝组织病理改变;ELISA法检测肝脏TNFα水平;免疫组化检测各组大鼠肝组织中核因子κB蛋白表达情况。 结果 :蜕皮甾酮治疗组血清胆固醇(TC)、丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)明显低于模型组(212±058比263±024,Plt;005;5336±1848比8460±3627,P＜005;14020±3595比24359±3638,P＜001);蜕皮甾酮治疗组与模型组相比肝组织丙二醛(MDA)水平降低明显(18454±1645比23928±2376,P＜001),超氧化物歧化酶(SOD)活力增加显著(942±052比518±043,P＜001),肝脏指数显著降低(435±037比504±046,P＜001)，肝组织脂肪变性程度和炎症活动度明显减轻(546±037比630±049,P＜001)。蜕皮甾酮治疗组与模型组相比TNFα与核因子κB水平明显减轻(4304±748比6156±727,2465±539比4504±746,P值均＜001)。 结论 :蜕皮甾酮具有改善高脂饮食诱发的非酒精性脂肪性肝病大鼠肝脏酶学功能,通过增加肝组织SOD的含量和减少MDA的含量来减轻肝组织氧化应激水平,减轻肝组织TNFα和核因子κB来减轻肝脏炎症,发挥防治非酒精性脂肪性肝病的作用。Abstract: Objective: To investigate the effect and possible mechanism of ecdysterone on the expression of tumor necrosis factoralpha (TNFα) and nuclear factor κ B (NFκB) in rats with nonalcoholic fatty liver disease of rats. Methods : A total of 36 male Sprague Dawley rats were randomly divided into two groups, who were fed with highfat diet (experimental group, n=24) and normal basic food (normal control, n=12) respectively. At the end of the 12th week, the experimental group was randomly divided into two subgroups: model group and ecdysterone group, each group contained 12 rats. From the 13th week, the rats in the normal control group and model group were lavaged with normal sodium, and the rats in the ecdysterone group were lavaged with ecdysterone at 10 mg·kg-1·d-1. At the end of the 16th week, all rats were weighed, narcotized, sacrificed, and the liver index, biochemical indicators in serum and liver tissues and the hepatic pathological changes were observed. The expression of TNFα was detected by ELISA and the expression of NFκB was measured by immunohistochemical staining. Results : At the end 16th week in ecdysterone group, the serum levels of cholesterol (TC), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were reduced markedly (212±058 vs 263±024 and 5336±1848 vs 8460±3627, both P＜005; 14020±3595 vs 24359±3638, P＜001); the tissue content of malondialdehyde (MDA) was decreased evidently (18454±1645 vs 23928±2376, P＜001), while the activity of superoxide dismutase (SOD) was enhanced notably (942±052 vs 518±043, P＜001); the liver index was decreased significantly in comparison with that inmodel group (435±037 vs 504±046, P＜001); the degree of fatty degeneration and inflammation were relieved dramatically (546±037 vs 630±049, P＜001). The expression of TNFα and the levels of NFκB were significantly lower (4304±748 vs 6156±727 and 2465±539 vs 4504±746, both P＜001) in ecdysterone group compared with model group. Conclusion : The effects of ecdysterone in preventing NAFLD in rats could be related to the increase of SOD content in hepatic tissue and the decrease of MDA content, tumor necrosis factorα and NFκB.
Anti-tumor necrosis factor-α monoclonal antibody agents have been widely applied in the management of autoimmune diseases. Among them, Adalimumab and Infliximab have been used for years in clinical practice in treating non-infectious uveitis and achieved satisfactory effects and safety. However, no guideline or expert consensus for their usage is available in China currently. It hopefully promotes standardized clinical application of anti-tumor necrosis factor -α monoclonal antibody in treating non-infectious uveitis, together with other senior experts in uveitis, the Ocular Immunology Group of Immunology and Rheumatology Academy in Cross-Straits Medicine Exchange Association form this evidence-based recommendations for clinicians’ reference.
Objective To investigate the effects of one-lung ventilation time on the concentration of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in the bronchoalveolar lavage fluid (BALF), serum inflammatory markers and early pulmonary infection after radical resection of esophageal cancer. Methods Ninety patients with thoracoscope and laparoscopic radical resection of esophageal carcinoma were chosen. According to the thoracoscope operation time, the patients were divided into 3 groups including a T1 (0.5–1.5 hours) group, a T2 (1.5–2.5 hours) group and a T3 (>2.5 hours) group. Immediately after the operation, the ventilated and collapsed BALF were taken. Enzyme-linked immunosorbent assay (ELISA) method was used to determine the concentration of IL-6 and tumour necrosis TNF-α. The concentrations of procalcitonin (PCT), C-reactive protein (CRP), and white blood cell (WBC) were measured on the first, third, fifth day after operation. The incidence of pulmonary infection was observed within 3 days after operation. Result The IL-6 values of the right collapsed lung in all groups were higher than those in the left ventilated lung. The TNF-α value of the right collapsed lung in the T2 group and T3 group was higher than that in the left ventilated lung (P<0.05). Compared with in the right collapsed lung, the TNF-α and IL-6 values gradually increased with the the duration of one-lung ventilation (P<0.05). Compared with the left ventilated lung groups, the IL-6 value increased gradually with the duration of one-lung ventilation time (P<0.05). The TNF-α value of the T3 group was higher than that of the T1 and T2 groups (P<0.05). The PCT value of the T3 group was higher than that of the T1 group and T2 group on the third, fifth day after operation (P<0.05). But there was no significant difference in CRP and WBC among the three groups at different time points. The incidence of pulmonary infection in the T3 group was significantly higher than that in the T1 group within 3 days after operation (P<0.05). Conclusion With the extension of one-lung ventilation time, the release of local and systemic inflammatory mediators is increased, and the probability of pulmonary infection is higher.
Objective To study the variety and the action of inflammatory cytokines and the relevant anti-inflammatory factors in acute pancreatitis (AP). Methods The authors observed the change of peripheral blood IL-6 and sTNFR in 41 patients with mild and severe AP in two groups on 1, 5, 14d after acute attack by ELISA. Results All cases recovered gradually in mild group (n＝22) after five days. Twelve patients improved gradually in severe group (n＝19) after 5-7 days. The level of sTNFR increased markedly in 2 groups at 1, 5, 14d(Ｐ＜0.001), and that of the severe group was markedly higher decreased gradually (Ｐ＜0.01). The level of IL-6 increased apparently only in severe group on 1d, 40.38 pg/ml∶12.4 pg/ml, (Ｐ＜0.001). The levels of IL-6 and sTNFR correlated respectively with severity of AP. Conclusion These results show that peripheral blood IL-6 and TNFα are useful index to supervise the severity and conversion and final results of AP.
Objective To summarize and evaluate the significance of tumor necrosis factor (TNF) tolerance, and to understand the general characteristics and known molecular mechanisms of different forms of tolerance, including the roles of transcription factors, signaling systems and receptors. Method The relevant literatures at home and abroad in recent years were collected and readed, and the content related to TNF tolerance was summarized. ResultsTNF tolerance could be produced after TNF pretreatment, and be divided into absolute tolerance and induced tolerance. TNF tolerance was related to a variety of interrelated and interdependent intracellular signal transduction. There was cross tolerance between TNF and lipopolysaccharide. Conclusions TNF tolerance may represent a protective mechanism, participating in the termination of inflammation and preventing excessive or persistent inflammation. TNF tolerance may also trigger immune paralysis, leading to severe inflammatory diseases, such as sepsis. The understanding of TNF tolerance can promote the diagnosis of inflammation related diseases or the implementation of treatment methods, so as to achieve more accurate evaluation and treatment.
The tyrosine kinase activity of epidermal growth factor receptor (EGFR) plays a key role in tumor cell proliferation, invasion, migration, and drug resistance. Studies have shown that non-small cell lung cancer patients with somatic driver gene EGFR mutations are sensitive to and can benefit from EGFR-tyrosine kinase inhibitors (EGFR-TKIs). Nevertheless, EGFR-TKIs-related adverse events should not be ignored. Common adverse events such as diarrhea, acne-like rash and paronychia are usually manageable; although the incidence of interstitial lung disease is low, once it occurs, it is a serious threat to patients' life, and its pathogenesis is still unclear. There is very limited animal experimental and clinical research evidence on the potential mechanism of EGFR-TKIs-related interstitial lung disease in the available literature. Based on this, this article reviews the association between EGFR-TKIs and interstitial lung disease, at the same time, also discusses the research progress of EGFR-TKIs-related interstitial lung disease in combination with cytotoxic drugs or immunotherapeutic drugs and EGFR-TKIs, in order to provide a reference for the prevention and treatment of EGFR-TKIs-related interstitial lung disease in clinical practice in the future.
ObjectiveTo construct a predictive model for acute kidney injury (AKI) after coronary artery bypass grafting (CABG) based on uromodulin (UMOD) and tumor necrosis factor receptor-associated factor 6 (TRAF6). MethodsPatients undergoing CABG treatment at Tianjin Chest Hospital from 2022 to 2024 were prospectively enrolled. Based on whether they developed AKI post-surgery, patients were divided into the an AKI group and a non-AKI group. Differences in UMOD, TRAF6, blood urea nitrogen (BUN), serum creatinine (SCr), β-N-acetylglucosaminidase (NAG), and SCr clearance rate at different time points were compared between the two groups. Predictive models for AKI after CABG were constructed at various time points, and the predictive efficacy of the models for AKI was analyzed. ResultsA total of 70 patients were included, with 22 in the AKI group [13 males and 9 females, aged 55-72 (67.91±4.91) years] and 48 in the non-AKI group [32 males and 16 females, aged 56-72 (68.07±4.67) years]. The UMOD levels in the AKI group were lower than those in the non-AKI group at various time points including before surgery (t=34.283, P<0.001), postoperative 2 h (t=29.590, P<0.001), 4 h (t=30.705, P<0.001), 8 h (t=26.620, P<0.001), 12 h (t=29.671, P<0.001), and 24 h (t=31.397, P<0.001). The TRAF6 levels in the AKI group were higher than those in the non-AKI group at all these time points (P<0.001). Multivariate analysis showed that higher levels of TRAF6, BUN, SCr, NAG, and lower levels of UMOD and SCr clearance rate were risk factors for AKI after CABG (P<0.05). The receiver operating characteristic curve analysis showed that the area under the curve of the predictive model at postoperative 12 h was significantly higher than that of the remaining models. The risk of AKI after CABG was: log (Y)=12.333−1.582×UMOD+1.270×TRAF6+1.356×BUN+1.356×SCr+1.355×NAG−1.254×SCr clearance rate. ConclusionIn the occurrence process of AKI after CABG, TRAF6 exacerbates renal injury by activating inflammatory signals and promoting cell apoptosis, while UMOD alleviates renal injury by regulating renal tubular function and protecting renal tubular epithelial cells. Through the simulation analysis of the two biomarkers combined with renal injury indicators at postoperative 12 h, the occurrence of AKI after CABG can be effectively predicted.