Components of drugs in acupoint sticking therapy and its mechanism of intervention on bronchial asthma based on UPLC-Q-TOF-MS combined with network pharmacology and experimental verification / 中国中药杂志

UPLC-Q-TOF-MS combined with network pharmacology and experimental verification was used to explore the mechanism of acupoint sticking therapy(AST) in the intervention of bronchial asthma(BA). The chemical components of Sinapis Semen, Cory-dalis Rhizoma, Kansui Radix, Asari Radix et Rhizoma, and Zingiberis Rhizoma Recens were retrieved from TCMSP as self-built database. The active components in AST drugs were analyzed by UPLC-Q-TOF-MS, and the targets were screened out in TCMSP and Swiss-TargetPrediction. Targets of BA were collected from GeneCards, and the intersection of active components and targets was obtained by Venny 2.1.0. The potential targets were imported into STRING and DAVID for PPI, GO, and KEGG analyses. The asthma model induced by house dust mite(HDM) was established in mice. The mechanism of AST on asthmatic mice was explored by pulmonary function, Western blot, and flow cytometry. The results indicated that 54 active components were obtained by UPLC-Q-TOF-MS and 162 potential targets were obtained from the intersection. The first 53 targets were selected as key targets. PPI, GO, and KEGG analyses showed that AST presumedly acted on SRC, PIK3 CA, and other targets through active components such as sinoacutine, sinapic acid, dihydrocapsaicin, and 6-gingerol and regulated PI3 K-AKT, ErbB, chemokine, sphingolipid, and other signaling pathways to intervene in the pathological mechanism of BA. AST can improve lung function, down-regulate the expression of PI3 K and p-AKT proteins in lung tissues, enhance the expression of PETN protein, and reduce the level of type Ⅱ innate immune cells(ILC2 s) in lung tissues of asthmatic mice. In conclusion, AST may inhibit ILC2 s by down-regulating the PI3 K-AKT pathway to relieve asthmatic airway inflammation and reduce airway hyperresponsiveness.

Construction and anti-tumor efficacy of a peptide vaccine that targets calcium-binding protein S100A9 / 药学学报

Calcium-binding protein S100A9 is closely related to inflammation and tumor invasion, and is one of the specific markers of myeloid-derived suppressor cells (MDSC). In this study, a recombinant polypeptide vaccine CTB-S100A9 targeting mouse calcium-binding protein S100A9 was constructed by fusion cholera toxin B subunit (CTB) with S100A9 gene. The CTB-S100A9 fusion protein was expressed in E coli. and purified by Ni+ affinity chromatography. Vaccinate the purified recombinant CTB-S100A9 protein supplemented with aluminum hydroxide adjuvant can break the autoimmune tolerance and produce high titer of S100A9 antibody in mice. Moreover, the S100A9 antibody produced by CTB-S100A9 vaccination is more specific and does not cross-react with S100A8. In the mouse 4T1 breast cancer model, CTB-S100A9 vaccination not only has significant tumor prevention effects, but also has significant tumor therapeutic effects. In addition, CTB-S100A9 significantly inhibited lung metastasis in 4T1 mice breast cancer model. Further analysis by flow cytometry showed that CTB-S100A9 vaccination can significantly reduce the tumor induced Treg cells and granulocyte-derived MDSC in 4T1 mice model, and reverse the tumor immunosuppressive environment, thereby promote the anti-tumor efficacy. The animal experiments in this study were carried out under the animal care guidelines approved by the Animal Ethics Committee of the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. This study shows that CTB-S100A9 is a good recombinant vaccine that targets the tumor immune-suppression environment and has great potential for the future clinical application.

STUDY ON THE PROTECTIVE EFFECT OF ORGANOSELENIUM FROM SE-ENRLED LACTOBACILLUS ON CHEMICAL LIVER INJURY AND ITS MECHANISM / 营养学报

Objective: To study the protective effect in CCl4-induced liver injury by organoselenium from Se-enriched lactobacillus. Methods: (1) In the first series, forty-five animals were randomly divided into control (C) group, CCl4 group, CCl4 plus organoselenium group (CCl4-Se group). The liver injury was induced by abdominal injection of CCl4 every other day for 4 w. Changes of GSH-Px, CAT and SOD activities as well as MDA content in liver were estimated in the 2nd and 4th week after CCl4 injection respectively. (2) In the second series, forty-eight mice were randomly divided into C group, CCl4 group, CCl4 plus low dose organoselenium group (CCl4-LSe group) and CCl4 plus high dose organoselenium group (CCl4-HSe group). Changes of hepatocyte [Ca2+]i in animals in every group were investigated by means of confocal laser microscope on the 4th and 8th day after CCl4 injection respectively. Results: During the entire experimental period, liver MDA of CCl4 group was markedly superior to that of C and CCl4-Se groups, and the level of latter two groups was very close. The GSH-Px and CAT activities were higher in CCl4-Se group than in CCl4 group,but lower than that of C group. There were higher SOD activities in C and CCl4-Se groups compared to that in CCl4 group though without obvious difference. Average fluorescence pixels of hepatocyte [Ca2+]i in CCl4 group was 2.8 and 5.5 times higher than that of group C in the 4th and 8th day respectively,while those in CCl4-Se groups were significantly lower than those of CCl4 group, and close to C group. Conclusions: Organoselenium from Se-enriched lactobacillus, can protect hepatocyte [Ca2+]i homeostasis by reducing lipid peroxidation after CCl4 exposure.