Professor Wang Shouchuan’s Experience And Mechanism Of Deficiency-excess Differentiation In The Treatment Of Infantile Viral Pneumonia

Objective:To explore Wang Shouchuan’s experience in the treatment of infantile viral pneumonia and its corresponding modern scientific mechanism.Methods:On the basis of the original text of classical ancient books of traditional Chinese medicine,this paper analyzes its strategy of tonifying and reducing deficiency and excess of asthma and cough in pneumonia.Then discusses the origin of Wang Shouchuan’s thought of replenishing and reducing deficiency and reducing deficiency and how to apply it to the prevention and treatment of children viral pneumonia.A mouse model of H1N1 pneumonia was established by nasal drip of influenza A virus(H1N1).Lung tissue pathology was evaluated by hematoxylin-eosin staining;Protein levels in mice alveolar lavage fluid(BALF)were determined by BCA method;The mRNA expressions of IL-6,TNF-α and IL-1β in the lungs of mice were detected by QPCR;Protein expressions of TNF-α and IL-1β in mice BALF were detected by ELISA;The protein expression of NLRP3 and caspasel-p20 in the lung of mice was assessed by immunohistochemistry;The expression of NLRP3 in CD45+ cells and CD45-cells was detected by flow cytometry;The alveolar macrophages(AM)in BALF were extracted from mice;The mRNA expressions of IL-1β and NLRP3 in mice AM were detected by QPCR,and the protein expressions of NLRP3 and caspasel-p20 in AM were detected by flow cytometry;Mice lung macrophages were removed with macrophage scavenger Clodronate nasal drip;Activation of NLRP3 was inhibited by MCC950,an inhibitor of NLRP3.The immunocompromised mouse model was established by intraperitoneal injection of dexamethasone;A mouse model of respiratory syncytial virus(RSV)pneumonia was established by nasal drip.The proportion of CD8+T cells,CD11c+ cells and NK cells in spleen,mesenteric lymph node and lung of mice was determined by flow cytometry;The proportion of MDSC,M-MDSC and G-MDSC in spleen and bone marrow of mice was determined by flow cytometry;The mRNA expressions of RSV-F,RSV-G and ISG-15 in the lungs of mice were detected by QPCR;Lung tissue pathology was evaluated by hematoxylin-eosin staining;The ratio of CD45+lymphocytes and CD8+CD69+T cells in mice BALF was determined by flow cytometry.Results:The clinical operation mode of selecting reinforcing and reducing methods according to the deficiency and reality of patients was formed in the Qin and Han dynasties,especially represented by Huangdi Neijing and Jingui Yaolue.In the Huangdi Neijing,under the pathological condition,"deficiency" refers to the weakness of vital qi,and "excess" refers to the exuberance of evil qi.Distinguishing deficiency and excess is used to guide the means of tonifying and reducing in drug treatment and acupuncture treatment,that is,"deficiency is tonifying it,and actually purging it",the means is relatively simple.In the synopsis of the Golden Chamber,the distinction between deficiency and excess is mainly used to guide the means of tonifying and reducing in drug treatment.It needs to be emphasized that the means of tonifying and reducing deficiency and excess in synopsis of the Golden Chamber are far from as simple as those in Neijing,and gradually form a set of programmed system.Professor Wang Shouchuan inherited the idea of dividing deficiency and excess into diseases in the classic works of Qin and Han dynasties,and creatively applied it to the diagnosis and treatment of infantile viral pneumonia.It is considered that in the initial stage of the disease,fever,cough,phlegm obstruction,shortness of breath and nasal fan are the main symptoms,and the treatment is mainly to open the lung and resolve phlegm,detoxify and activate blood circulation,and self-made Jinxin oral liquid as the basic prescription.For the susceptible children who may have recurrent respiratory tract infection or reinfection,it is considered that the current treatment of viral pneumonia can not be stopped,and the significance of follow-up treatment is to prevent reinfection.The basic pathogenesis of this part of children in remission period is that the lung defense is not solid,the camp and defense is not in harmony,the method should tonify the lung and stabilize the surface,reconcile the camp and defense,and set Jinping decoction as the basis for treatment.The two prescriptions,Jinxin oral liquid,dispelled evil and solid,treated the disease,and Jinping decoction rectified deficiency and treated no disease,which had a good clinical effect in the prevention and treatment of infantile viral pneumonia.Jinxin oral liquid can significantly alleviate the lung pathology of H1N1 infection by alleviating lung injury,inhibiting pulmonary inflammation and reducing the titer of H1N1.In this study,the treatment of Jinxin oral liquid could reduce the level of NLRP3 in the lung tissue of H1N1 model mice and reduce the secretion of IL-1β into the alveolar cavity,suggesting that the protective effect of Jinxin oral liquid on H1N1 infection may depend on NLRP3 inflammatory bodies.Therefore,we use MCC950,a specific inhibitor of NLRP3,to block the NLRP3 pathway in advance.After blocking the NLRP3 pathway,Jinxin oral liquid could not inhibit the production of IL-1β and lost its protective effect on lung pathology induced by H1N1.In addition,Jinxin oral liquid could significantly inhibit the activation of NLRP3 pathway in AM and the expression of IL-1β in AM.It is suggested that the inhibitory effect of Jinxin oral liquid on IL-1β induced by AM is necessary for the treatment of H IN 1 infected mice.In order to confirm the role of AM in the efficacy of Jinxin oral liquid,100 μL chlorophosphonate liposomes were administered intranasally to expel AM from the lungs.The absence of AM weakened the protective effect of Jinxin oral liquid.After the establishment of dexamethasone model group,the related indexes of mice in dexamethasone model group and normal group were detected,and the body weight,spleen weight and the number of immune cells(including CD8+T,NK,DC)in spleen and mesenteric lymph nodes were selected as evaluation indexes.It was found that the immunity of mice in dexamethasone model group decreased significantly,and the proportion of MDSC in bone marrow and spleen increased significantly in model group.After administration of Jinping decoction,the body weight,spleen weight and the number of immune cells(including CD8+T,NK,DC)in lungs and mesenteric lymph nodes were selected as evaluation indexes.It was found that the immunity of mice in Jinping decoction group increased significantly,and the proportion of MMDSC in bone marrow and spleen decreased significantly in Jinping decoction group.At the end of administration,the model was made by nasal drip of RSV,and the samples were taken to detect the related indexes one day later.It was found that compared with the model group,the load of RSV virus in the lungs of the Jinping decoction group was lower,and the pathological and inflammatory infiltration of the lungs was lighter.Further study found that Jinping decoction could increase the expression of type I interferon and the number of CTL in BALF of mice,suggesting that Jinping decoction may play an anti-RSV effect through type I interferon and CTL.In addition,the number of central memory T cells in PBMC increased significantly,suggesting that Jinping decoction may play a protective role in the event of RSV reinfection.Conclusion:(1)Professor Wang Shouchuan established Jinxin Oral liquid to treat infantile viral pneumonia and Jinping decoction to prevent recurrent or easy attacks of viral pneumonia in weak children.The two prescriptions have achieved good clinical results in the prevention and treatment of viral pneumonia in children.(2)Jinxin oral liquid improved the pulmonary inflammation caused by H1N1 by inhibiting the excessive activation of NLRP3 inflammatory bodies in alveolar macrophages.This mechanism may be corresponding to the empirical study of Jinxin oral liquid in the treatment of pneumonia.(3)Jinping decoction can significantly improve the immunity of immunocompromised mice induced by dexamethasone,and Jinping decoction can improve the pulmonary inflammation and reduce the viral load induced by RSV in immunocompromised mice.The mechanism may be related to the antiviral effect of type I interferon and CTL.This mechanism may be corresponding to the fact that Jinping decoction can prevent viral pneumonia infection in frail children.