Qingfei Tongluo Mixture Attenuates Bleomycin-Induced Pulmonary Inflammation and Fibrosis through mTOR-Dependent Autophagy in Rats.

As an interstitial fibrosis disease characterized by diffuse alveolitis and structural alveolar disorders, idiopathic pulmonary fibrosis (IPF) has high lethality but lacks limited therapeutic drugs. A hospital preparation used for the treatment of viral pneumonia, Qingfei Tongluo mixture (QFTL), is rumored to have protective effects against inflammatory and respiratory disease. This study aims to confirm whether it has a therapeutic effect on bleomycin-induced IPF in rats and to elucidate its mechanism of action. Male SD rats were randomly divided into the following groups: control, model, CQ + QFTL (84 mg/kg chloroquine (CQ) + 3.64 g/kg QFTL), QFTL-L, M, H (3.64, 7.28, and 14.56 g/kg, respectively) and pirfenidone (PFD 420 mg/kg). After induction modeling and drug intervention, blood samples and lung tissue were collected for further detection. Body weight and lung coefficient were examined, combined with hematoxylin and eosin (H&E) and Masson staining to observe lung tissue lesions. The enzyme-linked immunosorbent assay (ELISA) and the hydroxyproline (HYP) assay kit were used to detect changes in proinflammatory factors (transforming growth factor-ß (TGF-ß), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß)) and HYP. Immunohistochemistry and Western blotting were performed to observe changes in proteins related to pulmonary fibrosis (α-smooth muscle actin (α-SMA) and matrix metalloproteinase 12 (MMP12)) and autophagy (P62 and mechanistic target of rapamycin (mTOR)). Treatment with QFTL significantly improved the adverse effects of bleomycin on body weight, lung coefficient, and pathological changes. Then, QFTL reduced bleomycin-induced increases in proinflammatory mediators and HYP. The expression changes of pulmonary fibrosis and autophagy marker proteins are attenuated by QFTL. Furthermore, the autophagy inhibitor CQ significantly reversed the downward trend in HYP levels and α-SMA protein expression, which QFTL improved in BLM-induced pulmonary fibrosis rats. In conclusion, QFTL could effectively attenuate bleomycin-induced inflammation and pulmonary fibrosis through mTOR-dependent autophagy in rats. Therefore, QFTL has the potential to be an alternative treatment for IPF in clinical practice.

Wuweixiaoduyin regulates TAZ-mediated immunoregulatory properties of Treg/TH17 cells in chronic osteomyelitis.

Wuwei xiaoduyin (WWXDY) is a prescription for Chronic osteomyelitis (COM) in traditional Chinese medicine (TCM). However, its specific mechanism remains unclear. The objective of this study was to investigate the mechanism of WWXDY in COM treatment. To clarify the potential role of TAZ in the treatment of COM by WWXDY via regulatory CD4+ T cells differentiation. The expressions of TAZ, RORγt and Foxp3 were determined by Quantitative Real-time PCR and Western blot. Besides, levels of IL-21, IL-17 and IL-10 in peripheral blood were detected by using ELISA. Molecular dynamics simulations and docking were further utilized to explore the binding mechanism. COM resulted in abnormal cell differentiation and an imbalance of Treg/Th17. In comparison with the control group, the percentage of Treg cells, Foxp3 expression and secretion of IL-17 and -21 cytokines decreased (P < 0.001), while the proportion of Th17 cells, the levels of TAZ and RORγt and concentration of IL-10 in PBMCs increased in the COM group (P < 0.001). Furthermore, the above abnormal differentiation and function of Treg/Th17 cells in COM were suppressed after treatment with WWXDY in vivo and in vitro. In addition, TEAD1 inhibited the therapeutic effect of WWXDY in terms of Treg/Th17 cells with COM. it was found that the main active ingredients were cichoric acid and isocarlinoside. WWXDY regulates immunoregulatory properties of Treg/Th17 cells in COM mainly by mediating TAZ expression. By inhibiting the chronic inflammation in COM, WWXDY is potentially used to inhibit the progression of COM into bone tumors.

Comprehensive characterization and quantification of multiple components in Dan-Huang-Qu-Yu capsule using a multivariate data processing approach based on microwave-assisted extraction with UHPLC and Q Exactive quadrupole-orbitrap high-resolution mass spectrometry.

Dan-Huang-Qu-Yu capsule, a Chinese herbal medicine compound preparation, is widely used for chronic pelvic inflammatory disease. In this study, a rapid, selective, and sensitive microwave-assisted extraction ultra-high-performance liquid chromatography-Q Exactive quadrupole-orbitrap high-resolution mass spectrometry method was developed for analyzing its chemical compositions. A total of 85 compounds, including 22 flavonoids, 8 terpenoids, 5 quinones, 5 phthaleolactone, 23 organic acids, and 22 other compounds were identified from Dan-Huang-Qu-Yu capsule. Among them, 35 major compounds were unambiguously detected by comparing them with reference standards and selected as quality control markers, which were simultaneously determined in Dan-Huang-Qu-Yu capsule. The established method was successfully validated and applied for simultaneous determination of 35 bioactive compounds in Dan-Huang-Qu-Yu capsule from ten sample batches. The quantitative data of the analytes were analyzed by principal component analysis for quality assessment of Dan-Huang-Qu-Yu capsule. Six compounds (e.g., astragaloside IV, salvianolic acid B, ellagic acid, chlorogenic acid, N-butylidenephthalide, and luteolin) were screened out and regarded as chemical markers for quality control of Dan-Huang-Qu-Yu capsule. The established method has been proved to be a novel and useful tool for rapid research of Dan-Huang-Qu-Yu capsule. This research will provide reference for the scientific research of traditional Chinese medicines.