Heme oxygenase-1 reduces inflammatory response by inhibiting thioredoxin interacting protein/NOD-like receptor protein 3 inflammasome activation in RAW264.7 cells / 中华危重病急救医学

Objective:To investigate the inhibitory effect and mechanism of heme oxygenase-1 (HO-1) on the inflammatory response of macrophages.Methods:Mouse macrophage strain RAW264.7 was cultured in vitro, and the cells in the logarithmic growth phase were used for the experiment. The RAW264.7 cells were divided into four groups. In blank control group, the cells were continuously incubated and received no treatment (cultured at 37 ℃, 95% air, 5% CO 2). In lipopolysaccharide (LPS) model group, 1 mg/L LPS was added to the medium to prepare LPS challenge model. In HO-1 inducer group, the cells were incubated with 30 μmol/L HO-1 inducer hemin for 1 hour, and then 1 mg/L LPS was added for incubation. In HO-1 inhibition group, the cells were incubated with 5 μmol/L HO-1 specific antagonist Zinc protoporphyrin Ⅸ (ZnPPⅨ) for 0.5 hour, and then 1 mg/L LPS was added for incubation. After 48 hours of incubation with LPS, the supernatant of each group was taken, and the protein expressions of HO-1, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3) and mitochondrial autophagy marker microtubule-associated protein 1 light chain 3B (LC-3B) were detected by Western blotting. The expression of reactive oxygen species (ROS) was detected by immunofluorescence staining. Results:Compared with the blank control group, the cells in the LPS model group had a certain stress response, and autophagy occurred in mitochondria, but the expression of some inflammatory factors was restricted, which was related to the impairment of cell function. The protein expressions of HO-1, IL-1β, LC-3B, ROS were significantly increased, the protein expressions of TNF-α, TXNIP, and NLRP3 were decreased significantly, indicating that the cells were seriously injured after LPS challenge, and the model was successfully established. Compared with the LPS model group, HO-1 protein expression in the HO-1 inducer group was significantly increased (HO-1/GAPDH: 0.31±0.03 vs. 0.22±0.03, P < 0.05), the protein expressions of TNF-α, IL-1β, TXNIP, NLRP3, LC-3B and ROS were significantly inhibited [TNF-α protein (TNF-α/GAPDH): 0.08±0.01 vs. 0.45±0.05, IL-1β protein (IL-1β/GAPDH): 0.50±0.01 vs. 0.82±0.03, TXNIP protein (TXNIP/GAPDH): 0.21±0.02 vs. 0.28±0.02, NLRP3 protein (NLRP3/GAPDH): 0.11±0.01 vs. 0.17±0.02, LC-3B protein (LC-3B/GAPDH): 0.67±0.04 vs. 0.92±0.12, ROS (fluorescence intensity): 80.9±12.5 vs. 94.1±19.5, all P < 0.05], indicating that HO-1 could inhibit inflammatory response and oxidative stress, and reduce mitochondrial autophagy. Antagonizing HO-1 could increase inflammatory response, oxidative stress and mitochondrial autophagy, the inhibitory degree of TNF-α and IL-1β expression was significantly reduced as compared with the HO-1 inducer group [TNF-α protein (TNF-α/GAPDH): 0.26±0.02 vs. 0.08±0.01, IL-1β protein (IL-1β/GAPDH): 0.76±0.01 vs. 0.50±0.01, both P < 0.05], the protein expressions of TXNIP, NLRP3, LC-3B and ROS were significantly increased as compared with the LPS model group [TXNIP protein (TXNIP/GAPDH): 0.43±0.02 vs. 0.28±0.02, NLRP3 protein (NLRP3/GAPDH): 0.24±0.02 vs. 0.17±0.02, LC-3B protein (LC-3B/GAPDH): 1.12±0.07 vs. 0.92±0.12, ROS (fluorescence intensity): 112.0±17.0 vs. 94.1±19.5, all P < 0.05]. Conclusion:HO-1 can reduce the inflammatory response by inhibiting the activation of TXNIP/NLRP3 inflammasome and reducing the release of inflammatory mediators.

Efficacy and adverse reaction of azithromycin sequential therapy for the treatment of mycoplasma pneumonia in children / 中国基层医药

Objective To investigate the efficacy and adverse reactions of azithromycin sequential therapy for the treatment of children with mycoplasma pneumonia.Methods 84 children with mycoplasma pneumonia were randomly divided into study group and control group according to the digital table,with 42 cases in each group.The study group was given azithromycin treatment,and the control group was given erythromycin treatment.The clinical curative effect and adverse reaction were compared between the two groups.Results The total effective rate of the study group (97.62％) was significantly higher than that of the control group (76.19％),and the difference was statistically significant (x2 =11.95,P ＜ 0.05).The pyretolysis time [(3.78 ± 1.57) d],cough improvement time [(3.29 ± 0.74) d],rate disappearance time[(5.88 ± 2.35) d],average hospitalization time [(8.89 ± 1.76) d] of the study group were significantly shorter than those of the control group [(4.49 ± 1.71) d,(4.28 ± 0.77) d,(7.69 ±2.54) d,(10.05 ± 2.27) d],and the differences were statistically significant (t =2.01,6.33,3.29,2.46,all P ＜0.05).The incidence rate of adverse reactions such as nausea,abdominal distention,abdominal discomfort,local injection pain,rash,elevated alanine aminotransferase of the study group (4.76％) was significantly lower than that of the control group (19.05％),and the difference was statistically significant (x2 =4.86,P ＜ 0.05).Conclusion Azithromycin sequential therapy is effective in the treatment of children with mycoplasma pneumonia.It can effectively shorten the disappearance time of clinical symptoms and signs,and average hospitalization time,and has low incidence rate of adverse reactions.It is worthy of clinical use.

A new anthraquinone from roots of Rumex japonicus / 中国中药杂志

A new anthraquinone, (1)-hydroxymethyl-3,6-dimethoxyl-2,8-dihydroxylanthraquinone 1, was isolated from the root of Rumex japonicus along with six known compounds 2-7. Their structures were elucidated by various spectroscopic methods including 2D-NMR techniques or comparison with authentic samples.

Chemical constituents of the bark of Celastrus orbiculatus / 中成药

AIM:To study the chemical constitutes of Celastrus orbiculatus Thunb. METHODS:Root of celastmus orbiculatus was macerated with cool ethanol to obtain the extract under the vaccum drying processing,extract obtained was extracted respectively by petroleum ester and ethyl acetate. Final extracts were purified by Silicagel column and analyzed by UV,IR,Ms and NMR to identify the chemical structural formula. RESULTS:Seven compounds were isolated from Celastrus orbiculatus Thunb,from which five constituents came from petroleum ester extract,and identified as sugiol (1),pristimerin (2),?-sitosterol (3),celastrol (4),from which two constituents came from ethyl acetate extract,salapermic acid(5),benzoic acid (6) and daucosterol (7),respectively. CONCLUSION:Compounds 1,5 were obtained from the plant for the first time.