The Study Of Yin Qiao San On The Mouse Model Of Respiratory Diseases Infected With Respiratory Syncytial Virus

Respiratory syncytial virus (respiratory syncytial virus, RSV) is a major worldwide public health problem. RSV infection is one of the main factors leading to the infection of the lower respiratory tract infections in infants, bronchiolitis and pneumonia in the most important pathogens, and also of children worldwide epidemic respiratory disease.RSV infection is still no convincing safe vaccine against the virus, there is no effective antiviral treatment. Selection of currently available clinical treatment of RSV drugs less, has been the treatment of RSV infection drug Western medicine mainly antivirals, bronchodilators, steroid anti-inflammatory drugs and immunomodulatory agents. But these drugs have serious side effects exists, but the vaccine has to be ineffective. Compared with Western medicine, Chinese medicine side effects of small, rich source of medicine, prevention and treatment of RSV infection has a unique effect. Advantage of Chinese medicine to prevent RSV infection is to adjust the body’s immune status, to enhance the body’s immunity against influenza virus. Therefore, the search for effective drugs to prevent RSV infection in the treasure house of traditional Chinese medicine has become a work of great significance. Yinqiaosan of traditional Chinese medicine, Chinese medicine Xin Liang Xie table method of party representatives, is the clinical syndrome of wind-heat treatment of influenza major prescription. There are practical efficacy of influenza, pneumonia, bronchitis, tonsillitis, mumps and other disease areas are Chinese warmed viral infection diseases. The formula by the forsythia, honeysuckle, bitter Campanulaceae, mint, leaves, raw licorice, Nepeta ear, SSP, Arctium, phragmites 10 kinds of pharmaceutical composition. Among them, honeysuckle, forsythia both Xinliang thoroughly evil, the power of the heat, but also with aromatic provision of foul detoxification effect. Mint, Arctium and throat to Xinliang of sexual dispersing heat. Nepeta ear, SSP is Xin Wen of herbs help to open fur and by evil, provision of foul aroma. Bamboo can be clear on hot coke; phragmites can heat fluid; bellflower can Xuanfei cough. Licorice can reconcile the various drugs, safety in Huwei, but also co bellflower clearing the throat. Studies have shown that the various components Yinqiaosan points are able to inhibit RSV infection may play a role.This study established in vivo mouse model of RSV infection in the respiratory system and in vitro RSV infected epithelial cells TC-1 mouse model, the use of low, medium and high dose Yinqiaosan (respectively 150 mg/kg,300 mg/kg and 600mg/kg) and ribavirin (150mg/kg) were treated by lung index was measured, HE pathological lung tissue and nasal tissue inflammatory infiltration staining change, detection of inflammatory cytokines IL-1β\TNF-a and the expression of IL-6 from animal and cellular levels observed Yinqiaosan on RSV-induced respiratory infections whether an improved effect. RSV infection often leads to immune injury and inflammation occurs. In the process of activation of inflammatory cytokines, NALP3 inflammatory body has an indispensable role. NALP3 inflammasome nucleotide domain-like receptors (Nalps) family, the immune system of pathogen receptors can recognize the pathogen and generates a corresponding immune response. NALP3 inflammatory body by the NOD-like receptor family member 3 (NALP3), apoptosis-associated speck-like protein (ASC), and caspase enzyme caspase-1 composition. When the ligand binding NALP3 promote the formation of inflammatory corpuscles, and subjected to self-activate caspase-1, resulting in maturation and secretion of various inflammatory factors IL-1β and IL-18, etc., caused by a variety of immune and inflammatory responses.After this study established in vivo model of RSV infection of the respiratory system and give Yinqiaosan intervention, using qPCR and western blot detection NALP3 inflammatory-associated protein NALP3, ASC and caspase-1 expression, and to explore the treatment of RSV infection Yinqiaosan effect and NALP3 inflammatory activation of relations in order to further reveal Yinqiaosan against RSV infection effect an immune response signaling mechanism for RSV infection clinical application provide a theoretical basis for further.ObjectiveTo investigate the effect and regulating mechanism of Yin Qiao San on the mouse model of respiratory diseases infected with respiratory syncytial virus (RSV).MethodsSection 1:Mouse model of respiratory diseases was constructed by infected with RSV. Yin Qiao San at low (150mg/kg), middle (300 mg/kg) and high (600 mg/kg) dose or ribavirin (150mg/kg) were used to treat mouse model of respiratory diseases. Lung index were counted by measure the weight of body and lung; The morphologic change such as nasal mucosa and inflammatory cell infiltration was detected by Hematoxylin and Eosin (HE) staining; The expression levels of IL-1β, TNF-a, IL-6 in bronchoalveolar lavage fluid were detected by enzyme-linked immunosorbent assay (ELISA); The expression levels of NALP3, ASC and caspase-1 in lung tissues were detected byreal-time quantitative PCR and western blot.Section 2:Mouse lung epithelial cell line TC-1 cells were infected with respiratory syncytial virus, and were treated with Yin Qiao San, ribavirin or transfected with NALP3 siRNAto suppress the expression of NALP3. All group cells were cultured for 2 days. The expression levels of IL-ip, TNF-a and IL-6 in cellular supernatant were ELISA; The expression levels of NALP3, ASC and caspase-1 in each group cells were detected byreal-time quantitative PCR and western blot.ResultsSection 1:The results in vivo revealed that Yin Qiao San could reduce lung index, suppress the expression of NALP3, ASC and caspase-1, suppress the formation of NALP3inflammasome, and suppress the expression of IL-1β, TNF-a, and IL-6 in mouse model of respiratory diseases infected with RSV. The results of HE staining showed that Yin Qiao San could reduce the injury of nasal mucosa and lung. The therapeutic effect of high and middle dose Yin Qiao San is similar to ribavirin.Section 2:Cell assays in vitro revealed that Yin Qiao San could suppress the expression of NALP3, ASC and caspase-1, suppress the formation of NALP3inflammasome, and suppress the expression of IL-1β, TNF-a, and IL-6. The effect of high and middle dose Yin Qiao San is similar to ribavirin.ConelusionYin Qiao San could reduce the expression of NALP3, the formation of inflammasome and the secretion of inflammatory cytokines to suppress the infection of RSV. The above conclusion might be one of mechanism that Yin Qiao San treats RSV infection. Our study suggested using high and middle dose Yin Qiao San to treat RSV infection.