| Objective:1.To observe the clinical efficacy of Huangqi Huayin granules in the treatment of AECOPD.2.To explore the main active components and potential mechanisms of Huangqihuayin granules in the treatment of acute exacerbation of chronic obstructive pulmonary disease(AECOPD)through network pharmacology of traditional Chinese medicine compound,and to screen the core targets and related signaling pathways.Based on PI3K-Akt1-Nrf2 signaling pathway,animal experiments were conducted to verify the effects of Huangqihuayin granules on inflammation and oxidative stress in AECOPD rats.Methods:1.Part of clinical study: 110 patients with AECOPD who met the diagnostic criteria were randomly divided into treatment group and control group.Both groups were treated with basic treatment,the treatment group was treated with Huangqihuayin granules,and the control group was treated with Guilong Kechuanning capsules.The changes of TCM major and minor symptom scores,6-minute walk test,random blood oxygen saturation,CAT score,dyspnea score,BODE index,IL-6,IL-1β,high-sensitivity C-reactive protein,white blood cell count,neutrophil count,lymphocyte count and lung function were observed before and after treatment in the two groups.The statistical tool software SPSS(software version 26.0)was used to analyze the data to observe the safety and effectiveness.2.Network Pharmacology: TCMSP database was used to collect the active ingredients and action targets of the drugs contained in Huangqi Huayin granules.Drug Bank,Gene Cards,OMIM and other databases were used to collect the genes related to AECOPD.Cytoscape3.7.2 software was used to establish the network diagram of drug-ingredients-target information.The String platform was used to make the PPI network diagram of the intersection targets of drugs and diseases,and the core targets were predicted.3.Animal experiment: The rat model was established by cigarette smoke exposure combined with tracheal instillation of lipopolysaccharide.SD male rats were randomly divided into normal group,model group,low dose of Huangqi Huayin granule group,middle dose of Huangqi Huayin granule group,high dose of Huangqi Huayin granule group and roxithromycin group.HE staining and Masson staining were used to observe the pathological changes of rat lung tissue under a microscope.The expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),superoxide dismutase(SOD),Nrf2 and hs-CRP in serum and bronchoalveolar lavage fluid(BALF)were determined by enzyme-linked immunosorbent assay(ELISA).The expression levels of PI3 K m RNA,Akt1 m RNA,Nrf2 m RNA,HO-1m RNA,and Keap1 m RNA were detected by RT-q PCR.The expression of P-PI3 K,PI3K,PAkt1,Akt1,Nrf2,HO-1 and Keap1 in PI3K-AKt1-Nrf2 signaling pathway was detected by Western-bolt.Results:1.The comparison between the two groups before treatment showed that there was no obvious abnormality in the results(P > 0.05),which was comparable.(1)The results of TCM syndrome scores before and after treatment showed that the curative effect of the treatment group was significantly higher than that of the control group,the effective rate of the treatment group was 92%,and the effective rate of the treatment group was 94%,and the scores of the treatment group were better than those of the control group.(2)The 6-minute walk test(6MWT)was significantly increased in both groups before and after treatment(P < 0.01),but was better in the treatment group than in the control group.The blood oxygen saturation in the treatment group(P < 0.01)was higher than that in the control group(P < 0.05).The CAT score,m MRC score and BODE index of the two groups were significantly decreased before and after treatment(P < 0.01).(3)After treatment,the levels of IL-6,IL-1β,hs-CRP and white blood cell count in the treatment group were significantly decreased(P < 0.01),while the neutrophil count and lymphocyte count did not change significantly(P > 0.05).However,only IL-1β in the control group decreased significantly(P < 0.01),and IL-6,hs-CRP,white blood cell count,neutrophil count,and lymphocyte count did not change significantly(P > 0.05).(4)Pulmonary function tests showed that FEV1,FVC,FEV1% in the treatment group were significantly increased(P < 0.01),while there was no significant change in the control group(P > 0.05).2.After screening,a total of 136 active ingredients and 271 targets of Huangqihuayin granules were obtained.After screening,1895 targets were obtained from the disease database,and 174 intersection targets of Huangqihuayin granules and AECOPD were found.The core targets were screened by PPI,including AKT,TNF,IL-6,etc.The PI3K-Akt signaling pathway was included by KEGG enrichment analysis.GO analysis yielded biological processes: response to hormones,cellular response to nitrogen compounds,and so on.Cellular components: membrane rafts,membrane microzones and other components.The molecular functions mainly include transcription factor binding and protein domain specific binding.3.HE staining and Masson staining were used to determine the pathological changes of the model group,which were consistent with the pathological changes of AECOPD.(1)ELISA results showed that both serum and alveolar lavage fluid,compared with the model group,There were significant differences in TNF-α,IL-6,SOD,Nrf2 and hs-CRP between the high-dose Huangqihuyin granule group and the roxithromycin group(P < 0.01).The contents of these indexes in the low,medium and high dose groups of roxithromycin showed a gradient distribution,and the high-dose group of traditional Chinese medicine showed the greatest change.(2)Compared with the model group,the white blood cell count in the highdose Huangqihuyin granule group and the roxithromycin group decreased,but the roxithromycin group(P < 0.01)was better than the high-dose Chinese medicine group(P <0.05).The neutrophil count in the two groups decreased significantly(P < 0.01),and the Chinese medicine treatment groups still showed a gradient distribution.However,there was no significant difference in lymphocyte count among the three groups(P > 0.05).(3)RTq PCR results showed that compared with the model group,the high-dose Huangqihuayin granule group and the roxithromycin group had significantly down-regulated PI3 K m RNA and Akt1 m RNA(P < 0.01).The down-regulation of Keap1 m RNA in the high-dose Huangqihuayin granule group(P < 0.05)was slightly worse than that in the roxithromycin group(P < 0.01).The two groups had significant up-regulation of Nrf2 m RNA and HO-1m RNA(P < 0.01),and the distribution of each treatment group was still gradient.(4)Westernbolt results showed that compared with the model group,the expression of p-PI3 K /PI3 K and Keap1 in high-dose Huangqihuayin granule group and roxithromycin group was significantly decreased(P < 0.01),and the expression of p-Akt1 /Akt1 in roxithromycin group was decreased(P < 0.05).The expression of Nrf2 and HO-1 in the high-dose Huangqihuayin granule group was significantly lower than that in the high-dose Huangqihuayin granule group(P < 0.01),and the expression of Nrf2 and HO-1 in the two groups was significantly higher than that in the high-dose Huangqihuayin granule group(P < 0.01).Conclusion:1.Huangqi Huayin granules has a certain clinical effect,which can reduce the symptoms of patients,improve the quality of life,increase the 6-minute walking distance,improve lung function to a certain extent,and reduce related inflammatory indicators.2.The core targets of Huangqihuayin granules in the treatment of AECOPD mainly include AKT,TNF,IL-6,etc.The mechanism of intervention may be related to PI3K-Akt signaling pathway.3.Huangqi Huayin granules can reduce inflammation and oxidative stress by inhibiting PI3K-Akt1-Nrf2 signaling pathway. |
PDF Full Text Request