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ObjectiveBased on tumor necrosis factor alpha （TNF-α）/tumor necrosis factor receptor 1 （TNFR1）/receptor-interacting protein kinases （RIPKs） signaling pathway， this paper aims to study the effect of modified Erchentang on inflammation in rats with chronic obstructive pulmonary disease （COPD） and explore its mechanism of action. MethodA total of 60 SD rats were randomly divided into normal group， model group， high， medium， and low-dose groups （20， 10， 5 g·kg-1·d-1） of modified Erchentang， and Xiaokechuan group （3.5 mL·kg-1·d-1）， with 10 rats in each group. The COPD rat model was established by cigarette smoke combined with lipopolysaccharide （LPS）. The normal group and model group were given the same amount of normal saline for 21 days by gavage administration. The contents of TNF-α and TNFR1 in bronchoalveolar lavage fluid （BALF） of rats were detected by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expressions of RIPK1， RIPK3， and mixed lineage kinase domain-like （MLKL） in the lung tissue. The protein expressions of RIPK1， RIPK3， and MLKL in the lung tissue were detected by Western blot. The pathological changes in lung tissue were observed by hematoxylin-eosin （HE） staining. ResultCompared with the normal group， the contents of TNF-α and TNFR1 in BALF of the model group were significantly increased （P<0.01）， and the mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were significantly increased （P<0.01）. Compared with the model group， the contents of TNF-α and TNFR1 in BALF of high， medium， and low-dose groups of modified Erchentang and Xiaokechuan group were decreased （P<0.01）. The mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were decreased to different degrees （P<0.05， P<0.01）. ConclusionModified Erchentang can effectively improve the inflammatory response of lung tissue in COPD rats， and the mechanism may be by inhibiting the activation of the TNF-α/TNFR1/RIPKs signaling pathway.
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Objective::To explore the therapeutic effect and mechanism of Chaige Qinlian Tang on pneumonia in young mice. Method::The pneumonia model was duplicated by slowly dripping Staphylococcus aureus into the nasal cavity of mice.After successful modeling, the mice were randomly divided into model group, clindamycin group, and high and low-dose Chaige Qinlian Tang groups, with sham operation group as negative control group.The rats were given 200 mg·kg-1 high-dose Chaige Qinlian Tang, 100 mg·kg-1 low-dose Chaige Qinlian Tang and 120 mg·kg-1 clindamycin.The mice were observed every day.Colonies were counted in the lungs of each group five days later.The expression levels of interleukin(IL)-16, tumor necrosis factor (TNF)-α in lung lavage fluid of each group were determined by enzyme linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression levels of IL-16, TNF-α in lung lavage fluid of each group.The expressions of tumor necrosis factor receptor (TNFR) 1, Caspase-3 and Caspase-7 in lung and the pathological changes of lung were observed. Result::Compared with the sham operation group, the respiratory state and the activity state of the model mice were worse, and the survival rate was higher in the high-dose Chaige Qinlian Tang group.Compared with the sham operation group, the pulmonary colony counts in the model group and treatment groups were increased, compared with the model group, the lung colony counts in clindamycin group and high-dose Chaige Qinlian Tang group were improved significantly (P<0.05, P<0.01). Compared with the control group, the expression levels of IL-16, TNF-α, TNFR1, Caspase-3, Caspase-7 mRNA and protein in the lung of model group and treatment groups were significantly increased (P<0.01). Compared with model group, the expression levels of IL-16, TNF-α and TNFR1, Caspase-3, Caspase-7 in the lung of clindamycin group and high and low-dose Chaige Qinlian Tang groups were significantly increased (P<0.01). The expression levels of protein and mRNA were significantly decreased (P<0.05, P<0.01), and the pathological changes of lung were improved, especially in clindamycin group and high-dose Chaige Qinlian Tang group. Conclusion::Chaige Qinlian Tang has a certain therapeutic effect on Staphylococcus aureus pneumonia in young mice.This effect may be related to regulating TNFR1, Caspase-3 and Caspase-7 pathways, reducing the secretion of IL-16 and TNF-alpha, and enhancing the clearance of staphylococcus aureus.
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Objective To observe the effect of signal transduction pathway of NF-κB on tubular cell apoptosis in ischemia-reperfusion induced acute kidney injury (AKI) in mice.Methods Eighteen C57B/6 mice were randomly (random number) divided into three groups,namely control group,AKI group,and pyrrolidine dithiocarbamate (PDTC) group.AKI model of mouse was made by occlusion of bilateral renal pedicles with microvascular clamps for 45 minutes,and intraperitoneal injection of PDTC (50 mg/kg) was given immediately after modeling in mice of PDTC group.Forty-eight hours after modeling,kidney pathological changes,serum creatinine (SCr) and blood urea nitrogen (BUN) were examined,and renal tissue NF-κB,TNFR,Bcl-2 and caspase-3 levels were detected by using immunohistochemistry,and tubular cell apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL).Results (1) The pathological Pallers score of renal damage,blood urea nitrogen and serum creatinine levels in PDTC group were significantly lower than those in AKI group [(2.83 ± 0.41)vs.(4.50± 0.55),P=0.000; (61.65 ±3.06) mmol/L vs.(77.78 ±5.82)mmol/L,P=0.000and (74.33 ± 9.83) μmol/L vs.(152.00 ± 16.55) μmol/L,P =0.000,respectively].(2) The level of NF-κB in renal tissue homogenates in PDTC group was significantly lower than that in AKI group [(20.33± 2.34) ％ vs.(35.83 ± 3.06) ％,P =0.000].(3) The apoptotic index of renal tubular cells in PDTC group was significantly lower than that in AKI group [(16.67 ± 1.15) ％ vs.(28.00 ±2.01) ％,P =0.001].(4) The levels of caspase-3 and TNFR1 in renal tissue homogenates in PDTC group were significantly lower than those in AKI group [(7.00 ± 1.26) vs.(11.00 ± 1.26),P =0.000 and (5.55 ± 0.82) vs.(9.75 ± 0.76),P =0.000],and Bcl-2 level in PDTC group was significantly higher than that in AKI group [(10.50± 1.38)vs.(1.83 ±0.98),P=0.000].Conclusions NF-κB activates renal tubular cell apoptosis in acute kidney injury induced in mice after ischemia-reperfusion.Blockade of NF-κB signal transduction pathway may lessen the apoptosis of renal tubular cells,leading to renal function less compromised.
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Objective:Comparing the expression of tumor necrosis factor receptor 1 (TNFR1) in decidua tissue and soluble tumor necrosis factor receptor 1(sTNFR1) in serum of normal pregnancy and spontaneous abortion mice to probe the relationship between TNFR1 and unexplained spontaneous abortion.Methods:The abortion-prone CBA?DBA/2 mating was established as the model of spontaneous abortion and nonabortion-prone CBA?BALB/c matings were used as the model of normal pregnancy.Immunohistochemistry method(SABC) was employed to detect the expression of TNFR1 in decidua tissue at the day 9 of gestation.The level of sTNFR1 in serum at the same time was determined by ABC-ELISA.Results:Compared with normal pregnancy model,the expression of TNFR1 in decidua tissue of spontaneous abortion was significantly increased (P