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《自然》子刊重磅研究!武汉病毒所证实:金银花可抑制新冠病毒复制

2020-08-07 前瞻网 前瞻网

此前,已有研究表明,双黄连口服液、金银花、绿茶能够防控新型冠状病毒感染,但有不少人称,这些研究仅属于初步研究,对于普通民众防疫来说没有太多参考价值。

此前,已有研究表明,双黄连口服液、金银花、绿茶能够防控新型冠状病毒感染,但有不少人称,这些研究仅属于初步研究,对于普通民众防疫来说没有太多参考价值。

近日,一项研究切实表明金银花汤中的植物MIR2911能够抑制SARS-CoV-2的复制,并且加速了新冠感染患者的痊愈。该研究成果发表在《自然》子刊《细胞发现》(CellDiscovery)上。

作者包括南京大学张辰宇教授团队、武汉病毒所张磊砢,及南京市第二医院易永祥等。

此前的研究表明,在金银花汤剂(HD)中富集的植物microRNAMIR2911,可以通过与甲型流感病毒(IAV)的mRNA结合并阻断蛋白翻译,直接靶向甲型流感病毒(IAV),包括H1N1、H5N1和H7N9亚型。

口服HD可预防IAV感染,减少H5N1诱导的小鼠死亡。

后续研究表明,除了IAVs,MIR2911还可以直接抑制多种病毒的复制。

在经过膳食摄取后,这些microRNAs会自我组装成外泌体,然后分泌到循环中,并运送到靶组织或特定细胞,包括肝、肺、脾、胰腺和T细胞。

鉴于MIR2911独特的富含GC的核苷酸组成,研究人员分析了SARS-CoV-2的基因组序列后,发现病毒基因组很可能含有MIR2911结合位点,MIR2911可以直接抑制SARS-CoV-2的复制。

在本研究中,他们评估了HD中吸收的MIR2911对SARS-CoV-2复制的抑制作用,并对HD在COVID-19患者中的疗效进行了临床研究。

通过生物信息学分析,他们预测在SARS-CoV-2基因组中有179个推定的mir2911结合位点。

经典荧光素酶实验确认了28个结合位点,它们广泛分布在病毒基因组中(Fig.1a),表明MIR2911可能能够抑制几乎所有SARS-CoV-2蛋白的翻译。

为了评估吸收的MIR2911对SARS-CoV-2复制的直接影响,从转染合成MIR2911或对照非编码RNA(ncRNA)的HEK293T细胞培养基中收集细胞外泌体,方法与之前的报道相似(Fig.1b)。

将分离的含或不含MIR2911的细胞外泌体与VeroE6细胞(ATCC-1586)分别在0.25ml细胞培养基中孵育8小时。

改变培养基后,细胞感染SARS-CoV-2,病毒感染复数(MOI)为0.01。

在感染后24小时(p.i),通过实时定量RT-PCR(qRT-PCR)对细胞培养液中的病毒拷贝数进行定量评估(Fig.1b)。

如图1c所示,细胞外泌体-MIR2911在13.2pM的浓度下抑制了93%的病毒复制,表明吸收的MIR2911直接并充分抑制了SARS-CoV-2的复制(Fig.1c)。

接下来,他们评估了在HD中吸收的MIR2911对SARS-CoV-2复制的抑制作用。

由于在HD中吸收的MIR2911通过循环由外泌体进入肺,分别从饮200mlHD(30g干金银花)前后的供者血清62.5mol/l中分离的外泌体分别与VeroE6细胞孵育,与细胞外泌体-MIR2911的情况相似(Fig.1b)。

200mlHD中MIR2911的浓度为52.5pM(10.5pmol/200ml/30g金银花干)。

三个健康接受者在饮用2小时后的血清MIR2911水平为0.42,0.45和1.13pM(Fig.1d),在饮前无法检测到。

如图1e所示,含有MIR2911的外泌体显著抑制了病毒复制(Fig.1e)。

从同一供体收集的MIR2911在饮用HD前后的外泌体细胞存活率没有差异。

这些结果强烈表明,饮用HD的人可以充分吸收MIR2911,并且吸收的MIR2911可以显著阻止SARS-CoV-2的复制。

为了评估HD中MIR2911对COVID-19患者的抗病毒作用,他们进行了一项临床研究。

本研究纳入2020年1月至2020年3月在南京第二医院接受常规抗病毒治疗的75例中重度COVID-19患者。

将患者在RT基础上加用MIR2911治疗(MIR2911+)或中药合剂(MIR2911-)分为两组,以第一次治疗后第7天的转化率为主要终点。

MIR2911+组和MIR2911-组分别为6例和69例。

MIR2911+组第7天的阴转化率为83.3%,较MIR2911-TCM组有明显改善(26.1%,P=0.004)(Fig.1g,f)。

接受HD-MIR2911治疗的患者,变得阴性(TTN)所需的时间也对他们有利。(平均4.0天和12.0天,HR0.28,95%CI0.12-0.67,P=0.005)(Fig.1g,f)

男性患者MIR2911+(1例)和MIR2911-(38例)的中位TTN分别为5.0天和11.0天(HR0.003,95%CI0.000018-0.52,P=0.027)。

女性患者MIR2911+组(5例)和MIR2911-组(31例)的中位TTN分别为3.0天和12.0天(HR0.15,95%CI0.031-0.68,P=0.014)。

在中国,HD被用于治疗病毒感染已有一千年的历史。

既往研究表明,1-3mlHD中的MIR2911(0.06-0.18pmol/day)在20g小鼠中显著抑制流感病毒复制。

他们选择30克金银花干(MIR2911水平:10.5pmol)每天使用。

结果表明,30克金银花干使用是安全的,具有充分的抗病毒功能(Fig.1d,e)。

另一方面,数据显示外泌体中的MIR2911显著抑制病毒复制,不仅证实了MIR2911超强的抗病毒活性(remdesivir:3.7μM和氯喹:10μM),还提供了一个新策略,使用从健康供体收集的血清外泌体提供了最相似的体内条件来评估潜在药物在体外的疗效。

综上所述,结果表明,在HD中吸收的植物MIR2911抑制了SARS-CoV-2的复制,并加速了感染患者的负转化。

HD治疗可能极大地帮助治愈感染患者,阻止COVID-19大流行。

原文来源:

https://www.nature.com/articles/s41421-020-00197-3

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    2020-08-07 公卫新人

    新冠肺炎,疫情何时才能消失

    0

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