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Objective To explore the impact of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury(VILI)through adenylate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)signaling pathway.Methods Sixty SPF male rats were randomly grouped into the sham operation group,the model group,the low-dose Jiawei Taoren Chengqi Decoction group(TCM-L group,2.85 g/kg),the high-dose Jiawei Taoren Chengqi Decoction group(TCM-H group,8.55 g/kg)and the high-dose Jiawei Taoren Chengqi Decoction + AMPK inhibitor Compound C group(TCM-H+CC group,Jiawei Taoren Chengqi Decoction 8.55 g/kg+ Compound C 250μg/kg),12 in each group.Oxygenation index(OI)was measured immediately after intubation and at 1 h,2 h and 4 h after mechanical ventilation.Bronchoalveolar lavage fluid(BALF)was collected after mechanical ventilation,levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β and IL-18 in BALF were detected by enzyme-linked immunosorbent assay(ELISA).Rats were sacrificed,and lung tissue was taken to measure the wet/dry weight(W/D)ratio.HE staining was used to observe pathological changes of lung tissue in each group of rats.Lung injury scores were carried out.Morphology of alveolar epithelial cells was observed by transmission electron microscopy.RT-qPCR was applied to detect mRNA expression levels of AMPK and mTORC1 in rat lung tissue.Western blot assay was applied to detect expression levels of AMPK,p-AMPK,mTORC1,p-mTORC1 and autophagy-related proteins in rat lung tissue.Results Compared with the sham group,the pathological damage of lung tissue was serious,lung W/D,levels of TNF-α,IL-1β and IL-18 in BALF,lung injury score,mTORC1 mRNA expression level,and p-mTORC1 protein expression were increased in the model group(P＜0.05).OI values at 2 h and 4 h of mechanical ventilation,AMPK mRNA expression level,p-AMPK,LC3-II/LC3-I and Beclin-1 protein expression in lung tissue were decreased(P＜0.05).Compared with the model group,the pathological damage of lung tissue was alleviated in the Chinese medicine-H group,and the trend of changes in related indexes was opposite to the above(P＜0.05).Autophagosomes in alveolar epithelial cells were increased.Compound C attenuated the protective effect of Jiawei Taoren Chengqi Decoction on VILI rats(P＜0.05).Conclusion Jiawei Taoren Chengqi Decoction may promote autophagy and reduce VILI in rats by activating AMPK/mTOR signaling pathway.
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To explore the role of miR-873 in cardiomyocyte injury induces by hypoxia reoxygenation (H/R) and its related mechanisms. Methods: H/R model was established by culturing mouse cardiac H9c2 cells in vitro, and miR-873 mimic was transfected. The experiments were divided into a control group, a H/R group, a negative control group and a miR-873 mimic group. The expression of miR-873 was measured using real-time PCR. The protein expression levels of egl-9 family hypoxia inducible factor 3 (Egln3), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) were evaluated by Western blotting. Cell apoptosis ELISA kit and cysteine-containing, aspartate-specific proteases-3 (caspase-3) activity kit was used to detect cell apoptosis and caspase-3 activity, respectively. The targeting effect of miR-873 on Egln3 were examined by the dual luciferase report gene assay, and the experiments were divided into a negative control group, a Egln3 3'-untranslated regions (3'-UTR) WT group (WT group) and a Egln3 3'-UTR MUT group (MUT group). In order to further detect the effects of Egln3 on miR-873 mimics, the Egln3 overexpressed cells were constructed, and the experiments were divided into a H/R group, a H/R+miR-873 mimic group, a H/R+pcDNA3-Egln3 (pcEgln3) group and a H/R+ miR-873 mimic+pcEgln3 group. Results: Compared with the control group, the expression level of miR-873 was significantly decreased in the H/R group (P0.05). In the over-expression experiment, compared with the H/R group, the cell apoptosis and the ratio of Bax/Bcl-2 were significantly reduced in the miR-873 mimic group (both P<0.05). Compared with miR-873 mimic group, the cell apoptosis and the ratio of Bax/Bcl-2 were significantly up-regulated in the H/R+pcEgln3 group and the H/R+miR-873 mimic+pcEgln3 group (all P<0.05). Conclusion: MiR-873 can inhibit H/R- induced apoptosis of cardiomyocyte via targeting Egln3.
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Animals , Mice , Apoptosis , Cell Hypoxia , MicroRNAs , Myocytes, CardiacABSTRACT
Objective:To investigate the clinical application value of ultrasound in the diagnosis and differential diagnosis of Takayasu arteritis (TA ). Methods: A total of 39 suspected TA patients were examined by ultrasonography,erythrocyte sedimentation rate (ESR),C reactive protein (CRP)and CT angiography (CTA), and the ultrasound examination results were compared and analyzed with the final chinical diagnosis results. Results:Of the 39 cases of suspected TA patients which had been examined by ultrasound,33 cases were diagnosed as TA,2 cases were diagnosed as atherosclerosis (AS),2 cases were diagnosed as fibromuscular dysplasia (FMD) and 2 cases were diagnosed as thromboangiitis obliterans (TAO).Compared with final chinical diagnosis results, 1 FMD patient was misdiagnosed as TA,and the rest of the diagnostic results were consistent with the final chinical diagnosis results.Conclusion:Ultrasound examination can provide objective and accurate basis for the diagnosis and differential diagnosis of TA,and it is an effective method of imaging examination.