连花清瘟颗粒对新型冠状病毒肺炎的临床疗效及其机制的网络药理学研究

目的:探讨连花清瘟颗粒对新型冠状病毒肺炎(COVID-19)的临床疗效及其机制的网络药理学研究。方法:选取2020年1月20日~2020年2月28日在安徽医科大学附属六安医院入住的,均给予连花清瘟颗粒联合常规治疗的55例确诊为COVID-19的患者作为研究对象,统计分析其治疗前后主要临床症状改善情况及实验室检查指标的变化。通过TCM@Taiwan、TCMSP及TCMID数据库筛选的主要活性成分化合物及其主要靶点,采用GeneCards、OMIM及UniProt从人类基因数据库中筛选出与COVID-19相关的基因靶点,通过Cytoscape软件绘制治疗COVID-19的复方调控网络,并采用Cytoscape软件中自带Bisogenet程序构建治疗COVID-19的PPI蛋白互作网络,采用CytoNCA对PPI进行网络拓扑分析,最后采用R语言中Bioconductor程序包对筛选出的基因靶点进行GO和KEGG通路分析。结果:治疗后3 d,总有效率为74.55%,痊愈28人,患者主要体征包括体温、呼吸频率、脉搏均明显下降(P<0.05),治疗后7 d,总有效率为92.73%,痊愈39人,患者主要症状包括发热、咳嗽、乏力、胸闷人数相较于治疗前均明显减少(P<0.05或P<0.01),住院天数为(16.27±5.88)d,且通过统计核酸检测转阴时间为(14.98±5.27)天,患者住院期间未见与连花清瘟颗粒相关的不良反应及毒副作用。治疗前COVID-19患者淋巴细胞百分率有所降低,C反应蛋白明显升高,治疗后3 d,实验室检查正常率除CRP外均在80%以上,治疗后7 d,实验室检查正常率均在90%以上,且经CT影像证实好转49人,好转率89.09%。以网络药理学筛选到152个主要活性化合物,涉及到作用于新型冠状病毒肺炎的基因靶点有52个。GO富集分析和KEGG通路分析结果显示,主要涉及对脂多糖的应答、对细菌起源分子应答、对金属离子的反应及细胞生物刺激等生物过程,通过调节AGE-RAGE、TNF、卡波西氏肉瘤相关疱疹病毒感染、IL-17和人巨细胞病毒感染等信号通路作用于新型冠状病毒肺炎的治疗。结论:本研究通过临床观察,证实连花清瘟颗粒治疗COVID-19疗效确切,以网络药理学构建了连花清瘟颗粒作用于新型冠状病毒肺炎多活性成分、多基因靶点、多治疗通路的作用特点,较为全面地筛选出了连花清瘟颗粒作用于新型冠状病毒肺炎的基因靶点和信号通路,为深入探讨连花清瘟颗粒治疗新型冠状病毒肺炎的作用机制提供参考依据。

Clinical Efficacy of Lianhua Qingwen Granules and its Mechanism on COVID-19 Based on Network Pharmacology

Objective: To investigate the clinical efficacy of Lianhua Qingwen Granules on novel coronavirus pneumonia(COVID-19), and study its mechanism by network pharmacology. Methods: Fifty-five patients diagnosed as COVID-19 in Lu’an Hospital of Anhui Medical University from January 20, 2020 to February 28, 2020, were treated with Lianhua Qingwen Granules combined with conventional treatment. Improvements of main clinical symptoms and changes in laboratory indicators before and after treatment were analyzed. The main active ingredient compounds and their main targets were screened by using the TCM @ Taiwan, TCMSP and TCMID databases. GeneCards, OMIM and UniProt were used to screen COVID-19-related gene targets from the human gene database. Cytoscape software was used to draw a compound regulatory network for treating COVID-19, Bisogenet in Cytoscape software was used to construct a PPI protein interaction network for treating COVID-19, then CytoNCA was used to analyze the network topology of the PPI. Finally, the Bioconductor package in R language was used to analyze the GO and KEGG pathways of the selected gene targets. Results: The total effective rate was 74.55%, and 28 patients were cured after treatment for 3 days. The patient’s main signs including body temperature, respiratory rate, and pulse had a downward trend(P<0.05). After treatment for 7 days, the total effective rate was 92.73% and 39 patients were cured. Compared with before treatment, the main symptoms of the patients including fever, cough, fatigue, and chest tightness were significantly reduced(P<0.05). The number of hospital stays was(16.27±5.88) days, and the negative time was detected by statistical nucleic acid detection in(14.98±5.27) days, no adverse reactions or toxic side effects related to Lianhua Qingwen Granules were seen during the hospitalization. The percentage of lymphocytes in patients with COVID-19 was decreased before treatment, and the level of C-reactive protein was significantly increased. After treatment for 3 days, the normal rates of laboratory tests were above 80% except CRP. 7 days after treatment, the normal rates of laboratory tests were all above 90%, and 49 patients showed improvement with CT images, the improvement rate was 89.09%. A total of 152 active compounds were screened, and 52 genes were involved in the role of new-type coronary disease pneumonia. The GO enrichment analysis and KEGG pathway analysis results showed that it mainly involved biological processes such as the response to lipopolysaccharide, the molecular response to bacterial origin, the response to metal ions, and cell biological stimulation. Signaling pathways such as AGE-RAGE, TNF, sarcoma-associated herpesvirus infection, IL-17, and human cytomegalovirus infection affect the treatment of new-type coronary disease pneumonia. Conclusion: In this study, by using network pharmacology, the characteristics of Lianhua Qingwen Granules acting on multiple active components-multigene targets-multiple therapeutic pathways of new coronary pneumonia are established. The gene targets and signal pathways of the new type of coronary disease pneumonia are screened, which provide a reference basis for the in-depth study of the mechanism of Lianhua Qingwen Granule in the treatment of new type of coronary disease pneumonia.