Study On The Mechanism Of Rhubarb One-drug Dual-purpose Intervention On LPS-induced Pneumonia-related Inflammatory Factors In Mice

Purpose : Based on the network pharmacology method and the medicinal theory of traditional Chinese medicine,to explore the possible mechanism and material basis of rhubarb in the treatment of childhood pneumonia,and to establish a connection with the traditional Chinese medicinal theory;to establish a mouse model of pneumonia induced by lipopolysaccharide(LPS)inhalation and use Intervene with internal and external treatment methods of rhubarb.On this basis,determine the serum levels of interleukin 4(IL-4),interleukin 33(IL-4)and tumor necrosis factor(TNF-α)in mice,and observe the pathological morphology of mouse lung tissue And lung CT changes,to explore the common effect indicators of the anti-inflammatory effects of different therapies of rhubarb,and provide data support for the clinical treatment of pneumonia in children with one drug and dual use of rhubarb.Material and method:1 Screen and collect potential active ingredients and component targets of rhubarb through the Chinese Medicine System Pharmacology Database and Platform(TCMSP).Use Gene Cards and OMIM database platform to integrate the relevant targets of pediatric pneumonia,and combine with the component targets to obtain the key target of rhubarb in the treatment of pediatric pneumonia.Use String database combined with Cytoscape software to construct and analyze the protein interaction network of key targets,use Glue GO plug-in to analyze GO and KEGG energy enrichment,explore the mechanism of rhubarb in the treatment of pneumonia in children,and explain the network pharmacological results in combination with the medicinal properties of Chinese medicine.Traditional Chinese medicine theory establishes a connection with modern scientific methods.2 Screen 108 SPF mice and randomly divide them into normal group,control group 1,control group 2,internal treatment group,external treatment group,and dexamethasone group,with18 mice in each group.Except for the normal group,mice in the other groups were all established mouse acute lung injury models by LPS inhalation method.The treatment was started 24 hours after the model was established.The control group 1 mice were given normal saline by gavage,and the control group 2 mice were given corn flour and externally applied to the Backshu point.The internal treatment group and the external treatment group were respectively given rhubarb alcohol extract water solution and rhubarb alcohol by gavage.The cream was applied to the Backshu point,and the mice in the dexamethasone group were given intragastric administration of dexamethasone solution for 7 consecutive days,and lung CT examinations were performed before the first treatment and after the last treatment.On the 3rd,5th,and 7th day after the start of treatment,the mice of each group were sacrificed in batches to observe the pathological changes of the lung tissue of the mice;the lung index was determined by the weighing method;the IL-4 and IL-33 in the serum of the mice were detected by the Elisa method And TNF-α levels.Results:1.The results of network pharmacology show that the main active ingredients of rhubarb in the treatment of pediatric pneumonia include adenoflavin,rhein,cassia lactone,carotol,β-sitosterol,aloe-emodin,and catechins,which act on 36 Targets,the main core targets are MYC,JUN,and CASP3.By participating in the regulation of cell apoptosis,protein receptor binding,intervention of tumor necrosis factor pathway,trp inflammatory factor regulation pathway,etc.to treat pediatric pneumonia,it is in line with the bitter cold of rhubarb,clearing heat,and returning to the large intestine meridian.,Mainly purging the medicinal properties of traditional Chinese medicine for the treatment of pneumonia in children.2.General conditions: 108 experimental mice did not die during the experiment.The mice in the normal group were active and active,their fur was bright,and their mental state and diet were normal.The mice in the control group showed reduced activity 24 hours after the model was established.Hair is curly and lacklustre;poor mental state,reduced diet,and vertical hair;mice in each treatment group are generally better than the control group,and there is no significant difference between the groups.3.After HE staining,it was found that the lung tissue of the normal group of mice showed no abnormality;the lung tissue of the control group showed a large number of inflammatory cell infiltration,alveolar structure destruction and other pathological changes;the internal treatment group was compared with the dexamethasone group than the control group,The number of inflammatory cells was significantly improved,with only a small amount of inflammatory cell infiltration;compared with the control group,a small amount of inflammatory cells were seen in the external treatment group,accompanied by moderate blood stasis.After LPS induction,the lung index and pathological score of the mice in each group were higher than those in the normal group(P<0.05).After treatment,the internal treatment group,external treatment group and dexamethasone group were better than the control group(P<0.05).4.After induction of lipopolysaccharide,the lung CT of the mice in the normal group showed no abnormalities,and the lung texture was clear;the lungs of the mice in the other groups had fuzzy texture,disorder,or fuzzy patch shadow before treatment;On the 7th day after treatment,the above-mentioned performance of the mice in the control group was aggravated.The lung CT condition of the mice in the internal treatment group,the external treatment group and the dexamethasone group improved,but it was still different from the normal group;the results of CT parameters showed that the mice in the control group The lung volume decreased,the CT value increased,and the condition improved after treatment.There was no significant difference between the treatment groups(P<0.05).5.After lipopolysaccharide induction,the levels of IL-33 and TNF-α in the serum of mice in the control group increased,and reached the peak value on the 5th day after treatment.After drug intervention,the levels of IL-33 and TNF-α of mice in each treatment group were down-regulated,and the dexamethasone group had the best effect(P<0.05);IL-4 content increased on the 5th day(P<0.05),others there was no significant change at the time point(P>0.05).Conclusion:1.Rhubarb may have a regulatory effect on inflammation and cell apoptosis in the process of childhood pneumonia,and it is in line with the traditional Chinese medicine theory.2.Both internal and external use of rhubarb can effectively reduce LPS-induced lung tissue inflammation in mice.3.The therapeutic mechanism of rhubarb on LPS-induced pneumonia mice may be related to the regulation of the abnormal release of pro-inflammatory factors and the maintenance of the imbalance of the anti-inflammatory/pro-inflammatory factor ratio.4.IL-33 and TNF-α inflammatory factors can be used as the common effect index of internal treatment and external use of rhubarb.treatment and external use of rhubarb.