Mechanism of chlorogenic acid reducing lipopolysaccharide-induced acute lung injury in mice by regulating miR-223/NLRP3 axis / 中南大学学报(医学版)

OBJECTIVES@#Chlorogenic acid has various physiological activities such as antibacterial, anti-inflammatory, and antiviral activities. Studies have shown that chlorogenic acid can alleviate the inflammatory response of mice with acute lung injury (ALI), but the specific mechanism is still unclear. This study aims to investigate whether chlorogenic acid attenuates lipopolysaccharide (LPS)-induced ALI in mice by regulating the microRNA-223 (miR-223)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) axis.@*METHODS@#SPF grade BALBc male mice were randomly divided into a control group, a model group, a chlorogenic acid group, a chlorogenic acid+miR-223 negative control (miR-223 NC) group, and a chlorogenic acid+miR-223 inhibitor (miR-223 antagomir) group, 10 mice in each group. Except the control group, the other groups were instilled with 4 mg/kg LPS through the airway to establish the ALI mouse model. After the modeling, the mice in the chlorogenic acid group were continuously given chlorogenic acid (100 mg/kg) by gavage for 7 d. The chlorogenic acid+miR-223 NC group and the chlorogenic acid+miR-223 antagomir group were given 100 mg/kg chlorogenic acid by gavage every day, and then were injected with 10 μL of miR-223 NC (0.5 nmol/μL) and miR-223 antagomir (0.5 nmol/μL) respectively for 7 consecutive days.The control group and the model group were replaced with normal saline. The lung tissues of mice were taken to measure the ratios of lung wet to dry weight (W/D). The bronchoalveolar lavage fluid of mice was collected to measure the levels of TNF-α, IL-6, and IL-1β by ELISA kit and to count the number of eosinophils (EOS), lymphocytes, neutrophils under light microscope. After HE staining, the pathological changes of lung tissues were observed and lung injury was scored. qRT-PCR method were used to determine the expression levels of miR-223 in lung tissues. Western blotting was used to determine the expression levels of NLRP3 protein in mouse lung tissues. Luciferase reporter assay was used to analyze the targeting relationship of miR-223 to NLRP3.@*RESULTS@#Compared with the control group, the lung W/D value, the lung injury score and the level of inflammatory factors in the bronchoalveolar lavage fluid were significantly increased in the model group (all P<0.05); the infiltration of inflammatory cells in the lung tissue was severe; the alveolar space was significantly increased; the alveolar wall was significantly thickened; the number of EOS, lymphocytes, and neutrophils in the bronchoalveolar lavage fluid was significantly increased (all P<0.05); the expression levels of miR-223 in lung tissue were significantly decreased (P<0.05); and the protein expression levels of NLRP3 were significantly increased (P<0.05). Compared with the model group, the W/D value of lungs, lung injury score, and levels of inflammatory factors in bronchoalveolar lavage fluid were significantly decreased in the chlorogenic acid group, the chlorogenic acid+miR-223 NC group, and the chlorogenic acid+miR-223 antagomir group (all P<0.05); lung tissues damage was alleviated; the numbers of EOS, lymphocytes, and neutrophils in bronchoalveolar lavage fluid were significantly decreased (all P<0.05); the expression levels of miR-223 in lung tissues were significantly increased (P<0.05); and the expression levels of NLRP3 protein were significantly decreased (P<0.05). Compared with the chlorogenic acid group, the lung W/D value, lung injury score, and inflammatory factor levels in the bronchoalveolar lavage fluid were significantly increased in the chlorogenic acid+miR-223 antagomir group (all P<0.05); lung tissue damage was aggravated; the number of EOS, lymphocytes and neutrophils in bronchoalveolar lavage fluid significantly increased (all P<0.05); the expression levels of miR-223 in lung tissues were significantly decreased (P<0.05); and the expression levels of NLRP3 protein were significantly increased (P<0.05). The results of luciferase reporter assay showed that miR-223 had a targeting relationship with NLRP3.@*CONCLUSIONS@#Chlorogenic acid may increase the level of miR-223, target the inhibition of NLRP3 expression, reduce LPS-induced inflammatory response in ALI mice, and alleviate pathological damage of lung tissues.

Clinical significance of the monocyte to high-density lipoprotein cholesterol ratio in the assessment of imperfect ST-segment resolution in elderly patients with acute ST-elevation myocardial infarction after percutaneous coronary intervention / 中华老年医学杂志

Objective To investigate the clinical significance of the monocyte count/high-density lipoprotein cholesterol ratio(MHR)in evaluating imperfect ST-segment resolution in elderly patients with acute ST-elevation myocardial infarction(STEMI)after percutaneous coronary intervention(PCI).Methods This was a retrospective cohort study.A total of 274 elderly patients with STEMI underwent PCI in our hospital from December 2015 to December 2018 were enrolled.Based on the extent of the ST-segment resolution of the postoperative electrocardiogram,patients were divided into an imperfect ST-segment resolution group (observation group,n =79)and a favorable ST-segment resolution group (control group,n =195).General clinical data were compared between the two groups,and logistic regression equation was used to analyze the association of MHR with ST-segment resolution.Receiver operating characteristic(ROC)curve was performed to assess the predictive value of MHR for imperfect ST-segment resolution.Results Compared with patients in the control group,patients in the observation group were associated with a significantly higher proportion of anterior wall myocardial infarction and heart failure (≥ Killip 2),A longer duration of chest pain to balloon expansion,higher levels of creatine kinase isoenzyme,N-terminal pro-brain natriuretic peptide,hypersensitive C-reactive protein,blood sugar,blood uric acid,fibrinogen,triglyceride and mononuclear cell count,and lower levels of high density lipoprotein cholesterol and lymphocyte count (all P＜0.05).Meanwhile,there was a significant difference in MHR between the observation group and the control group [(0.75 ± 0.22) vs.(0.48 ± 0.19),t =9.831,P =0.001].Multivariate Logistic regression analysis showed that MHR was an independent risk factor for imperfect ST-segment resolution(OR =1.950,95％Cl:1.646-5.430,P =0.003)and ROC curve showed the threshold value of MHR at 0.67,the area under the curve at 0.867,the sensitivity at 79.72％,and the specificity at 79.61％.Conclusions MHR may be an independent risk factor and a good predictive index for imperfect ST-segment resolution in elderly patients with STEMI after PCI.

Clinical significance of the monocyte to high-density lipoprotein cholesterol ratio in the assessment of imperfect ST-segment resolution in elderly patients with acute ST-elevation myocardial infarction after percutaneous coronary intervention / 中华老年医学杂志

Objective@#To investigate the clinical significance of the monocyte count/high-density lipoprotein cholesterol ratio(MHR)in evaluating imperfect ST-segment resolution in elderly patients with acute ST-elevation myocardial infarction(STEMI)after percutaneous coronary intervention(PCI).@*Methods@#This was a retrospective cohort study.A total of 274 elderly patients with STEMI underwent PCI in our hospital from December 2015 to December 2018 were enrolled.Based on the extent of the ST-segment resolution of the postoperative electrocardiogram, patients were divided into an imperfect ST-segment resolution group(observation group, n=79)and a favorable ST-segment resolution group(control group, n=195). General clinical data were compared between the two groups, and logistic regression equation was used to analyze the association of MHR with ST-segment resolution.Receiver operating characteristic(ROC)curve was performed to assess the predictive value of MHR for imperfect ST-segment resolution.@*Results@#Compared with patients in the control group, patients in the observation group were associated with a significantly higher proportion of anterior wall myocardial infarction and heart failure(≥Killip 2), A longer duration of chest pain to balloon expansion, higher levels of creatine kinase isoenzyme, N-terminal pro-brain natriuretic peptide, hypersensitive C-reactive protein, blood sugar, blood uric acid, fibrinogen, triglyceride and mononuclear cell count, and lower levels of high density lipoprotein cholesterol and lymphocyte count(all P<0.05). Meanwhile, there was a significant difference in MHR between the observation group and the control group [(0.75±0.22)vs.(0.48±0.19), t=9.831, P=0.001]. Multivariate Logistic regression analysis showed that MHR was an independent risk factor for imperfect ST-segment resolution(OR=1.950, 95%CI: 1.646-5.430, P=0.003)and ROC curve showed the threshold value of MHR at 0.67, the area under the curve at 0.867, the sensitivity at 79.72%, and the specificity at 79.61%.@*Conclusions@#MHR may be an independent risk factor and a good predictive index for imperfect ST-segment resolution in elderly patients with STEMI after PCI.

Relationship Between Serum mRNA Level of Heparin Binding Epidermal Growth Factor and Acute Coronary Syndrome Occurrence / 中国循环杂志

Objective: To evaluate the relationship between serum mRNA level of heparin binding epidermal growth factor (HB-EGF) and acute coronary syndrome (ACS) occurrence. Methods: Our research included in 2 groups: ACS group,n=50 patients and Control group,n=100 normal subjects. Serum HB-EGF mRNA level was examined by RT-PCR and the relationship between HB-EGF mRNA and ACS occurrence was assessed by Logistic regression analysis. Results: Compared with Control group, the serum HB-EGF mRNA level of ACS group was higher (0.22±0.73) vs (0.46±0.14),P<0.05. With adjusted meaningful factors of hypertension, smoking, TG, TC, LDL-C, HDL-C and BMI by single factor analysis, multi Logistic regression analysis indicated that serum HB-EGF mRNA was related to ACS occurrence (OR=5.813, 95% CI 2.342-14.426,P<0.001) which meant that upon 0.1 grey value of HB-EGF mRNA elevation, the risk of ACS occurrence may increase 4.813 folds accordingly. Conclusion: Serum HB-EGF mRNA level was related to ACS occurrence.

Effects of crude antigen and excretory-secretory products of Clonorchis sinensis on T cells / 中国人兽共患病学报

We investigated the roles of the crude antigen(CA) of Clonorchis sinensis and excretory secretory products (ESPs) in the polarization of Th1 and Th2 cells.Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs;immature DCs were then treated with either CA (CA stimulated group),ESPs (ESPs stimulated group),LPS (positive control group) or PBS (negative control group) for 24 hours.Then the CD4+T cells were isolated from mouse spleen by using anti mouse-CD4 Microbeads,and further cocultured with stimulated DCs for another 72 hours.The purities of DCs and CD4+ T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3 mRNA were detected by real-time PCR.ELISA was used to detect the levels of IFN-γ and IL-4 cytokines in the supernatant.mRNA levels of T-bet and GATA-3 in the ESPs group were higher than those in PBS-stimulated group (P＜0.05).The concentrations of IFN-γ and IL-4 cytokines in the culture were increased in the ESPs group,compared with PBS stimulated group(P＜0.05).IFN-γ but not IL-4 was increased in CA group (P＜0.05).The results implied that CA might play a role in Th1 type immune response,and ESPs likely play roles in both Th1 and Th2 immune responses.