Neotuberostemonine and tuberostemonine ameliorate pulmonary fibrosis through suppressing TGF-β and SDF-1 secreted by macrophages and fibroblasts via the PI3K-dependent AKT and ERK pathways / 中国天然药物

Activated fibroblasts and M2-polarized macrophages may contribute to the progression of pulmonary fibrosis by forming a positive feedback loop. This study was aimed to investigate whether fibroblasts and macrophages form this loop by secreting SDF-1 and TGF-β and the impacts of neotuberostemonine (NTS) and tuberostemonine (TS). Mice were intratracheally injected with 3 U·kg-1 bleomycin and orally administered with 30 mg·kg-1 NTS or TS. Primary pulmonary fibroblasts (PFBs) and MH-S cells (alveolar macrophages) were used in vitro. The animal experiments showed that NTS and TS improved fibrosis related indicators, inhibited fibroblast activation and macrophage M2 polarization, and reduced the levels of TGF-β and SDF-1 in alveolar lavage fluid. Cell experiments showed that TGF-β1 may activated fibroblasts into myofibroblasts secreting SDF-1 by activating the PI3K/AKT/HIF-1α and PI3K/PAK/RAF/ERK/HIF-1α pathways. It was also found for the first time that SDF-1 was able to directly polarize macrophages into M2 phenotype secreting TGF-β through the same pathways as mentioned above. Moreover, the results of the cell coculture confirmed that fibroblasts and macrophages actually developed a feedback loop to promote fibrosis, and the secretion of TGF-β and SDF-1 was crucial for maintaining this loop. NTS and TS may disturb this loop through inhibiting both the PI3K/AKT/HIF-1α and PI3K/PAK/RAF/ERK/HIF-1α pathways to improve pulmonary fibrosis. NTS and TS are stereoisomeric alkaloids with pyrrole[1,2-a]azapine skeleton, and their effect on improving pulmonary fibrosis may be largely attributed to their parent nucleus. Moreover, this study found that inhibition of both the AKT and ERK pathways is essential for maximizing the improvement of pulmonary fibrosis.

Effect of intratracheal instillation of PM2.5 suspensionon pulmonary fibrosis in mice and the intervention of neotuberostemonine / 中国药科大学学报

@#To investigate the effects of intratracheal instillation of PM2.5 suspension on bleomycin (BLM)-induced pulmonary fibrosis in mice and the intervention of neotuberostemonine (NTS), the BLM dose (1.5 or 3.0 U/kg) and PM2.5 frequency (1 or 2 times per week) were studied by factorial experiment design. After intratracheal instillation of BLM (1.5 or 3.0 U/kg) on day 0, PM2.5 (5 mg/kg) was intratracheally injected to mice once or twice a week from day 1 to day 21, and the mice in the treatment group were given 30 mg/kg NTS by gavage once a day from day 8 to day 21. The degree of pulmonary fibrosis was evaluated by lung coefficient, hydroxyproline (HYP) content, HE staining and Masson staining lung sections as well as their semi-quantitative index (HE inflammatory score and collagen volume fraction, CVF). The results showed that the HE scores increased significantly in mice singly given PM2.5 once a week, the HYP content and HE score increased in mice singly given PM2.5 twice a week, but their CVF values did not significantly increase. However, the CVF values increased significantly in mice treated with PM2.5 and BLM co-infusion. These results suggested that PM2.5 (administered singly) could significantly increase BLM-induced collagen deposition and greatly aggravate pulmonary fibrosis although it mainly caused pulmonary inflammation rather than pulmonary fibrosis. NTS could significantly reduce the CVF value and α-SMA protein level of the model mice. It can be concluded that PM2.5 has great influence on patients with respiratory diseases, while NTS can improve pulmonary fibrosis induced by the combination of PM2.5 and BLM.

Simultaneous determination of ibuprofen and pseudoephedrin hydrochloride and chlorpheniramine maleate in the compound buluoweimanamin tablets by HPLC / 南方医科大学学报

<p><b>OBJECTIVE</b>An HPLC method was developed to determinate Ibuprofen and Pseudoephedrine Hydrochloride and Chlorpheniramine Maleate in Compound BuluoWeimaNamin Tablets.</p><p><b>METHODS</b>Using HPLC with Kromasil C18 column, and acetonitrile -0.5% SDS- phosphate (580:420:1) as the mobile phase. The wavelength for detection was 262 nm.</p><p><b>RESULTS</b>Better linearities and good correlation coefficients were obtained: the concentration ranges of ibuprofen, pseudoephedrine hydrochloride and chlorpheniramine maleate were over 2.062-14.434 microg (r=0.9999), 0.296-2.072l microg (r=0.9999), and 0.0204~0.1428 microg (r=0.9998), respectively. The recoveries of ibuprofen, pseudoephedrine hydrochloride and chlorpheniramine maleate were 99.98% (RSD=0.52%), 99.72 (RSD=0.82%) and 99.545 (RSD=0.76%), respectively.</p><p><b>CONCLUSION</b>The method was convenient, accurate and specific. It can be used as a method to control quality of Compound Buluoweimanamin Tablets.</p>

Studies on Apoptosis of Host Cells Induced by Foot-and-Mouth Disease Virus / 中国病毒学

Apoptosis of PK-15 cells induced by Foot-and-Mouth Disease Virus (FMDV) in vitro was reported in this paper. Typical cell apoptosis was detected by use of Hoechst 33258 fluorescence probe, agarose gel electrophoresis and in situ end-labeling (TUNEL). After PK-15 cells were infected by titration of 4.8 lg TCID50/mL FMDV for 32 h, apoptosis characteristics of nuclear condensation, fragmentation, accompanied by apoptotic bodies formation (Hoechst 33258 staining), 180-200 integer-fold sized pieces DNA Ladders (agarose gel electrophoresis) and strong green fluorescence dots (TUNEL) were all exhibited, and cell apoptosis was approximately 20%. In addition, the quantitative analysis of apoptosis in PK-15 cells induced by FMDV showed that apoptosis was correlated with infection of virus, and it was also time-dependent. Results indicate that FMDV can induce apoptosis of host cells and apoptosis plays an important role in the cytopathogencity effect of FMDV.