TRAF1 meditates lipopolysaccharide-induced acute lung injury by up regulating JNK activation.

Acute lung injury (ALI) is served as a severe life-threatening disease. However, the pathogenesis that contributes to ALI has not been fully understood. Tumor necrosis factor receptor-associated factor 1 (TRAF1) interacts with multiple regulators, performing its diverse role in biological functions. However, the effects of TRAF1 on ALI remain unknown. In this study, we attempted to explore the role of TRAF1 in ALI progression. The findings suggested that TRAF1-knockout (KO) markedly attenuated LPS-induced severe mortality rate in murine animals. LPS-elicited histological alterations in pulmonary tissues were significantly alleviated by TRAF1-deletion. Additionally, TRAF1 knockout effectively attenuated lung injury, as evidenced by the reduced lung wet/dry (W/D) weight ratio, as well as decreased bronchoalveolar lavage fluid (BALF) protein levels and neutrophil infiltration. Meanwhile, TRAF1 deletion markedly lessened inflammation, oxidative stress and apoptosis in BALF and/or lung tissues. The levels of pro-inflammatory cytokines stimulated by LPS were down-regulated by TRAF1 ablation, along with the inactivation of nuclear factor κB (NF-κB). LPS-promoted reactive oxygen species (ROS) generation was decreased in TRAF1-KO mice, partly through the improvement of anti-oxidants. Apoptosis was also inhibited by TRAF1 deletion in lung tissues of LPS-challenged mice through the suppression of cleaved Caspase-3. Moreover, TRAF1 knockout significantly decreased c-Jun N-terminal kinase (JNK) activation and its down-streaming signal of c-Jun in pulmonary samples of LPS-induced mice. Importantly, the in vitro study suggested that promoting JNK activation markedly abrogated TRAF1 knockdown-attenuated inflammation, ROS production and apoptosis in LPS-exposed A549 cells. Therefore, our experimental results provided evidence that TRAF1 suppression effectively protected LPS-induced ALI against inflammation, oxidative stress and apoptosis through the suppression of JNK activity.

Fingerprint study on Xiaochaihu Granules-medicinal materials based on peak pattern matching / 中草药

Objective: To establish Xiaochaihu Granules multi-wavelength fingerprints and conduct Bupleuri Radix-preparations peaks matching studies for further limitation validation of a single wavelength fingerprint quality evaluation. Methods: RP-HPLC was adopted to estimate the similarity on three batches of Xiaochaihu Granules from three manufacturers and three batches of Xiaochaihu Granules by laboratory-made (total 12 batches), and make peaks match raw herbs under the same conditions; the contents of indicator ingredients in different batches of Xiaochaihu Granules were determined by RP-HPLC method. Results: There was few differences of the fingerprint similarities among the three batches of Xiaochaihu Granules both from the same manufacture and laboratory-made at the wavelength of 210, 254, 280, and 323 nm. Among the fingerprints of Xiaochaihu Granules from different manufactures, the similarities of Xiaochaihu Granules only from manufacture A and of the three batches of Xiaochaihu Granules from the laboratory-made were over 0.9 under the four wavelengths. The similarities from the other two manufacturers were over 0.9 except 280 nm, but less than 0.9 at the wavelength of 210, 254, and 323 nm. At 210, 254, 280, and 323 nm wavelengths, peak pattern matching showed that Xiaochaihu Granules had 5, 5, 2, and 18 matching peaks with Bupleuri Radix, Glycyrrhizae Radix et Rhizoma, Codonopsis Radix, and Scutellariae Radix, respectively. The contents of glycyrrhizic acid and baicalin in Xiaochaihu Granules from different manufactures were significantly different. Saikosaponins a and d, ferulic acid, atractylenoide III, and lobetyolin were not detected in the formulations. Conclusion: There are great limitations to estimate quality of Xiaochaihu Granules with the fingerprint similarity at 280 nm. Multi-wavelength similarity evaluation combined with the peak matching of fingerprint for medicinal materials and preparations and quantitative determination of active components could provide a reference for the comprehensive quality evaluation of preparations for Chinese materia medica.