Prof. Chaobin Shen, Laboratory of Phytomedicine and Disease of Biomolecular, Department of Pediatrics, Shanghai TCM -Integrated Hospital, Shanghai, China
Biograph: CHAO-BIN SHEN is professor in pediatrics of Shanghai TCM-Integrated Hospital, Shanghai University of Traditional Chinese Medicine (China). He is graduate at pediatrics from Shanghai Second Medical College (at present is Medical College of Shanghai Jiao-Tong University) (1984). Actually is director of Pediatrics from Shanghai TCM-Integrated Hospital, At the moment is Fellow of Vaccine Council, Chinese Medical Association (CMA), Deputy Chairman of Academy of Pediatrics Immunology, Shanghai Medical Association (SMA), Chairman of Academy of Pediatrics Immunology and Rheumatology, Chinese Association of Integrated Medicine (CAIM), Deputy Chairman of Pediatrics Council, Shanghai Association of Integrated Medicine (SAIM), Fellow of American Academy of Immunologist (AAI). He has experience in clinical and basic immunology, epigenetic and traditional Chinese herbals. Chao-Bin Shen has been tutor of 36 medical master and doctor thesis and 6 medical master students, as an author of over than 70 scientific papers on Immunology and Chinese Herbals published in medical journals or presented at national and international congresses.

Speech Title: The inhibited the expression of GATA-3 on Th2 cells transfected Astragalus-derived miR-396 of asthmatic mice In vivo
Aim of the Study: MicroRNAs (miRNAs) are a class of small noncoding RNA that, through mediating posttranscriptional gene regulation, play a critical role in nearly all biological processes. Over the last decade it has become apparent that plant miRNAs may serve as a novel functional component of traditional herbs with therapeutic effects anti-inflammation and antitumor. Astragalus root has good properties in enhancing immune function as well as preventing and treating disease. In previous ours study, Next-generation sequencing identified miR-396 as the most abundant plant miRNAs expressed in Astragalus root, but the exact mechanism is unknown. Th2 cells are defined by their master transcription factor, GATA-3, and characteristic effector cytokine, IL-4, IL-5 and IL-13 in asthma model. Little is known about the functions of miR-396 in IL-5/IL-13-producing T cells. We investigated the effects of a synthetic miR-396 mimics (agomir) and its mechanism of inhibited action of Th2 cells of acute bronchial asthma in a mouse model.

Materials and Methods: BALB/c mice were sensitized and challenged with ovalbumin (OVA). The functional role of miR-396 was assessed by using in silico analysis. The miR-396 mimics was administered by intranasal instillation from the day of re-challenges every 2 days. In this study, mice of asthma model were fed synthetic miR-396 mimics in quantities of 200ng every mouse. On day 58, lung, spleen and blood were collected from the mice to examine, expression levels of miR-396 mimics in lung tissues, serums, and spleen were analyzed by RT-PCR. The cytokine levels (IFN-γ, IL-5, IL-13, IL-17A and TGF-β) in serum were detected by ELISA. The transcription factors mRNA (T-bet, GATA-3, STAT-6, ROR-γτ and Foxp-3) were determined by real-time PCR in mouse model with asthma and control groups in splenocytes. HE staining was used to observe the pathological changes of the lung tissue.

Results: In the present study, we show that administration of nasal drops containing miR-396 mimic was increased their expression significantly in lung tissues (10.69 times), spleens (13.87 times), and peripheral blood (12.62 times) from asthmatic mice after treatment compared with control mice. The miR-396 mimics suppressed the levels of Th2 cytokines, including IL-5 and IL-13 compared with that of the OVA-challenged groups (p＜0.001). GATA-3 of mRNA was significantly down-regulated (1.77 times) in asthma model of mice treated with miR-396 mimics in splenocytes. The miR-396 mimics also suppressed ROR-γt of mRNA compared with that of the OVA-challenged groups (3.33 times). We show that intranasal miR-396 administration reduces inflammation response in vivo in the lung’s pathological sections.

Conclusions: Our data suggest that miR-396 acts in a negative feedback loop to control the production of IL-5/IL-13 T cells and might thus impact the T-cell responses in asthma. We use qRT-PCR to confirm that intranasal delivery strategy efficiently in lung, serum and spleen tissue from mice with acute bronchial asthma. Our results suggest for the first time that plant miR-396 from Astragalus root of Chinese herbs plays a key role in regulating GATA-3 and ROR-γt expression and could be useful in developing better therapeutic strategies for alleviating lung inflammations.

Key words: Transfect miRNA in vivo; Astragalus; MicroRNA-396; Asthma; Animal model