百蕊颗粒抗小鼠流感病毒性肺炎作用及机制研究

目的研究百蕊颗粒对流感病毒致小鼠病毒性肺炎的影响及可能作用机制。方法将ICR小鼠按体重随机分组,分别为正常对照组,模型对照组,百蕊颗粒18.84、9.42、4.71 g·kg-1组,阳性对照药磷酸奥司他韦胶囊组。除正常对照组外,其余各组小鼠滴鼻感染流感病毒构建小鼠病毒性肺炎模型,测定小鼠的存活时间、死亡率;并测定病毒感染不同阶段(感染3、5、7 d后)正常对照组、模型对照组、百蕊颗粒18.84 g·kg-1组、磷酸奥司他韦胶囊组小鼠的体重、肺指数、肺组织病毒载量、血清炎症因子[白细胞介素-6(IL-6)、γ-干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)]水平。结果甲型流感病毒感染小鼠后,与正常对照组相比,模型对照组小鼠体重下降,肺指数、肺组织病毒载量、血清炎症因子水平升高,表明流感病毒性肺炎模型成立;与模型对照组相比,百蕊颗粒18.84、9.42、4.71 g·kg^(-1)组存活时间明显延长(P<0.01),死亡率明显降低(P<0.01),感染5、7 d后百蕊颗粒18.84 g·kg^(-1)组小鼠体重显著升高(P<0.05),感染3、5、7 d后百蕊颗粒18.84 g·kg^(-1)组小鼠肺指数值显著降低(P<0.05);感染5 d后百蕊颗粒18.84 g·kg^(-1)组病毒载量较模型组明显降低(P<0.01),感染7 d后百蕊颗粒18.84 g·kg^(-1)组血清IL-6含量较模型对照组显著降低(P<0.01)。结论百蕊颗粒对流感病毒感染致小鼠病毒性肺炎具有治疗作用,可提高病毒感染小鼠体重、降低肺指数,延长病毒感染小鼠存活时间、降低死亡率,其可能是通过降低肺组织病毒载量、血清炎症因子含量发挥抗流感作用。

当归水提取物补血、调节肠道菌群作用研究

目的:探究当归水提取物的补血作用,以及对血虚证模型小鼠肠道菌群结构的影响。方法:采用单纯放血法制备血虚证小鼠模型,通过测定小鼠外周血象、胸腺指数、红细胞膜Na^(+)-K^(+)-ATP酶活性评价当归水提取物的补血作用,并利用MiSeq平台对小鼠盲肠内容物进行16S rDNA测序,探究当归水提取物对小鼠肠道菌群结构的影响。结果:与模型组比较,当归组雌性小鼠外周血红细胞数量、白细胞数量显著升高(P<0.05),胸腺指数有上升趋势,红细胞膜Na^(+)-K^(+)-ATP酶活性显著升高(P<0.01);当归水提取物显著提高血虚证小鼠肠道菌群中的益生菌属如Lactobacillus(乳杆菌属)、Muribaculaceae(Muri菌)、Alloprevotella(拟普雷沃菌属)的丰度,且相关性分析表明,血虚证小鼠外周血红细胞数量、白细胞数量、血红蛋白量、血细胞比容与肠道多种微生物群落丰度具有显著相关性。结论:当归水提取物具有补血作用,且对小鼠肠道菌群具有调节作用。

基于转录组学的化湿败毒方治疗流感病毒性肺炎作用机制

目的:明确化湿败毒方对甲型H1N1流感病毒致小鼠肺炎的治疗作用,基于转录组学探讨其作用机制。方法:采用甲型H1N1流感病毒滴鼻感染建立小鼠流感病毒性肺炎模型,连续给药5 d,考察小鼠一般状况、肺指数、病毒载量、肺组织病理形态、生存时间、生存时间延长率等指标,明确化湿败毒方对流感病毒性肺炎的治疗作用。采用转录组学技术检测模型组与正常组、化湿败毒方组与模型组小鼠肺组织的差异表达基因,筛选化湿败毒方治疗流感病毒性肺炎的潜在核心靶点;采用实时荧光定量聚合酶链式反应(Real-time PCR)验证化湿败毒方对核心靶点基因mRNA表达水平的影响。结果:与正常组比较,模型组小鼠肺指数及肺组织内病毒载量均明显升高(P<0.05,P<0.01);与模型组比较,化湿败毒方高剂量组明显延长甲型流感病毒感染小鼠的生存时间(P<0.05),明显降低小鼠肺指数值(P<0.05),降低肺组织病毒载量,化湿败毒方高、中、低剂量组可明显减轻小鼠肺组织炎症、瘀血、肿胀等病变(P<0.05,P<0.01)。肺组织转录组学分析显示,化湿败毒方干预后核心基因主要富集在核转录因子-κB(NF-κB)信号通路、白细胞介素-17(IL-17)信号通路、细胞因子-细胞因子受体相互作用等通路,TNF受体相关因子6(TRAF6)、核转录因子-κB抑制因子α(NFKBIA)、趋化因子配体(CCL)2、CCL7、CXC趋化因子配体2(CXCL2)为combined score值前5的节点基因;Real-time PCR验证显示,化湿败毒方能明显下调NF-κB信号通路关键基因TRAF6、NFKBIA及趋化因子CCL2、CCL7、CXCL2 mRNA表达(P<0.05,P<0.01)。结论:化湿败毒方对流感病毒性肺炎具有治疗作用,可能通过抑制NF-κB信号通路关键节点TRAF6、NFKBIA表达,抑制趋化因子CCL2、CCL7、CXCL2的表达,减少炎症细胞的募集,降低病毒载量,发挥抗流感病毒肺炎作用。

Correlation between slow transit constipation and spleen Qi deficiency, and gut microbiota: a pilot study

OBJECTIVE:To investigate the effect of slow transit constipation(STC)and spleen Qi deficiency on gut microbiota,and the mechanism underlying the action that the positive drug Maren Runchang(MR)alleviates STC.METHODS:STC was induced,using the cathartic method of Senna and the hunger-fullness disorder method,in ICR mice;one group of model mice was treated with MR(6.24 g/kg).The changes in the general condition,fecal parameters,D-xylose content in the serum,intestinal propulsion rate,and histopathology of the colon were assessed after STC induction in the control,model,and MR groups.Fecal microbiota transplantation(FMT)was performed from STC mice into pseudo germ-free mice.Changes in the contents of substance P(SP),vasoactive intestinal peptide(VIP),and gut microbiota in STC mice and pseudo germ-free mice were assessed after FMT.RESULTS:Compared with the control group,the model mice showed the following results:the time of the first black stool was significantly longer(P<0.01),the number and weight of black stools were significantly reduced within 6 h(P<0.05),the D-xylose content in the serum was significantly reduced(P<0.05),the intestinal propulsion rate decreased(P<0.01),the content of VIP in colon tissue significantly increased(P<0.05),and SP content in the colon tissue significantly decreased(P<0.01);moreover,the colon showed significant inflamemation and injury.Furthermore,the abundance of Firmicutes was increased,the abundance of Bacteroides decreased,and the abundance of Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus decreased,while the abundance of the conditional pathogenic bacteria Escherichia and Klebsiella increased.However,after treatment with MR,the time of the first black stool decreased(P<0.01),the number of black stools within 6 h increased,and the intestinal propulsion rate increased(P<0.05).Moreover,the content of D-xylose in the serum and the content of VIP in colon tissue significantly decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and colon inflammation significantly improved.Additionally,the abundance of Firmicutes decreased,and the abundance of Bacteroides increased.The abundance of unclassified_o_Bacteroidales and Bilophila increased,and the abundance of Klebsiella decreased.In the model+FMT group,compared with control+FMT group,the content of VIP in colon tissue decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and the abundance of probiotics,such as Lactobacillus and Akkermansia,decreased.In the MR+FMT group,compared with the model+FMT group,the content of VIP in colon tissue increased,the content of SP in colon tissue significantly decreased(P<0.01),and the abundance of probiotics increased.CONCLUSIONS:STC mice with spleen Qi deficiency show a decreased abundance of beneficial bacteria,such as Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus,and an increased abundance of the conditional pathogenic bacteria Escherichia coli and Klebsiella.Furthermore,the mechanism of action of MR in treating STC may involve the regulation of intestinal movement,reduction of intestinal inflammation,elevation of intestinal absorption,and regulation of gut microbiota.

2022年国内外免疫学进展对中医药治疗COVID-19的启示

通过检索2022年国内外学术期刊发表的免疫学领域论文,综述其进展,重点介绍了天然免疫与炎症小体活化,T细胞免疫应答及免疫疗法,B细胞识别、调控及记忆功能,免疫耗竭,新型冠状病毒肺炎(COVID-19)免疫学机制及中医药防治进展。学科交叉与技术集成已成为推动免疫学发展的强大力量,随着高通量测序及筛选技术、全光谱流式细胞术、基因编辑技术、蛋白质工程技术、AI联合生物技术的发展及应用,研究人员得以深入挖掘天然免疫及适应性免疫的免疫应答特征、规律及其内在机制与靶点,从而寻找有效的治疗手段并开发针对适应症的疫苗及药物。2022年免疫学在基础理论及应用转化方面成果显著,但也面临着更大的挑战,新型冠状病毒的不断变异与其展现的免疫逃逸特点将持续性地困扰及危害人类社会,肿瘤及自身免疫疾病的免疫治疗如何提高其有效性与安全性,以及天然免疫与适应性免疫的基础研究还有待更深入的认识等。