WO2025001085A1 - Motor基因在制备用于治疗脓毒症的药物中的应用和药物 - Google Patents

Motor基因在制备用于治疗脓毒症的药物中的应用和药物 Download PDF

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WO2025001085A1
WO2025001085A1 PCT/CN2024/072889 CN2024072889W WO2025001085A1 WO 2025001085 A1 WO2025001085 A1 WO 2025001085A1 CN 2024072889 W CN2024072889 W CN 2024072889W WO 2025001085 A1 WO2025001085 A1 WO 2025001085A1
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motor
drug
gene
sepsis
cells
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黄蔚
邱海波
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谢剑锋
刘玲
杨毅
杨然
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Southeast University
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  • the present invention relates to the technical field of circRNA-encoded small peptide genes, and in particular to an application of a MOTOR gene in preparing a drug for treating sepsis and a drug.
  • Sepsis is a life-threatening organ dysfunction caused by the body's dysregulated response to infection, which poses a huge threat to human health.
  • a multicenter cross-sectional study in my country showed that sepsis patients accounted for about 20.6% of ICU hospitalizations in my country, and the 90-day mortality rate was as high as 35.5%. Therefore, in-depth research on the pathogenesis of sepsis and the search for new therapeutic target molecules are not only a major disease prevention and treatment need for the country and society, but also a huge challenge facing the clinic.
  • Immune imbalance is the fundamental mechanism of the occurrence and development of sepsis. Although multi-organ damage caused by excessive inflammatory response is an important part of the pathogenesis of sepsis, the hyperinflammatory state of sepsis is often accompanied by long-term and persistent immunosuppression. Monocyte LPS tolerance is a key feature of immunosuppression in sepsis. LPS-tolerant monocytes have a reduced ability to internalize and kill pathogenic microorganisms, and cannot respond effectively during secondary infection, which increases the risk of secondary infection in sepsis patients, and secondary infection is precisely the main cause of death in sepsis patients. However, the exact mechanism of LPS tolerance in monocytes has not yet been elucidated, and there is currently a lack of molecular drugs that regulate monocyte LPS tolerance. This is a core issue that urgently needs to be tackled.
  • the present invention proposes the use of the hsa_circRNA gene MOTOR in the preparation of a drug for the treatment of sepsis.
  • MOTOR reverses the LPS tolerance of sepsis monocytes and prolongs the survival time of mice.
  • the first aspect of the present application proposes the use of the MOTOR gene or its pharmaceutical derivatives in the preparation of drugs for treating sepsis, and the nucleotide sequence of the MOTOR gene is shown in SEQ NO.1.
  • the drug is used in a dosage form suitable for releasing the MOTOR gene or its pharmaceutically acceptable derivative in the user's peripheral blood.
  • the present application also provides a drug, which includes exosomes containing the MOTOR gene; the nucleotide sequence of the MOTOR gene is shown in SEQ NO.1.
  • the drug of the present invention can be added to conventional excipients and prepared into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral liquids, lozenges, injections, ointments, granules or various sustained-release preparations, according to conventional processes.
  • the carrier of the drug of the present invention is a common type available in the pharmaceutical field, including: a binder, a lubricant, a disintegrant, a solubilizer, a diluent, a stabilizer, a suspending agent or a matrix, etc.
  • the dosage form is an injection.
  • hsa_circRNA_056558 of the present application can reverse the LPS tolerance of monocytes during sepsis, promote the expression of chemokines, proinflammatory cytokines and antigen presentation genes secreted by monocytes under bacterial infection, regulate the innate immunity and adaptive immunity function of patients with sepsis, prevent secondary infection, and improve survival time.
  • FIG1 is a schematic diagram of the structure of the small peptide MOTOR located in the cytoplasm of the present application.
  • FIG2 is a diagram showing the inhibition of inflammatory factors and chemokines expression by knocking down MOTOR in the present application
  • FIG3 is a diagram showing that overexpression of MOTOR in the present application promotes the expression of inflammatory factors and chemokines
  • FIG. 4 is a particle size analysis diagram of the Motor exosomes of the present application.
  • the present invention uses the following steps to verify the effect of MOTOR on monocyte LPS tolerance during sepsis, and the specific steps are shown in Figures 1-4;
  • Example 1 MOTOR is a circRNA with coding ability, which can translate a small peptide of 137aa
  • MOTOR was identified as a circRNA by PCR and Sanger analysis.
  • the coding capacity of MOTOR was predicted by bioinformatics.
  • the coding capacity and subcellular localization of MOTOR were verified by Western blot and immunofluorescence techniques. It was found that MOTOR could translate into a small peptide localized in the cytoplasm (see Figure 1).
  • Reverse transcription PCR was performed on 500 ng of RNA using a reverse transcription kit (PrimeScript TM RT reagent Kit with gDNA Eraser, TAKARA, RR047A).
  • PCR primer sequences are as follows:
  • Flag-MOTOR THP1 cells were collected, fixed with 4% PFA at room temperature for 1 hour, rinsed with PBS three times, 5 minutes each time; blocked with Blocking medium (1% fetal bovine serum + 0.5% Tritonx-100) at room temperature for 1 hour, added with Flag antibody diluted 1:200 with PBS, and incubated overnight at 4°C.
  • Blocking medium 1% fetal bovine serum + 0.5% Tritonx-100
  • siRNA targeting MOTOR circRNA was ordered.
  • siRNA targeting MOTOR and control siRNA were transfected into cells using the NEON electroporation system. 1 ⁇ g/ml LPS was added 24 hours after transfection, and cells were harvested 24 hours after LPS treatment. The expression of cytokines and antigen presentation-related factors was detected by second-generation sequencing and qPCR, and it was found that knocking down MOTOR could significantly inhibit their expression (see Figure 2).
  • Trizol sample containing cells was quickly frozen in liquid nitrogen for 30 seconds, placed on dry ice and sent to the company for next-generation sequencing.
  • CT values obtained after qPCR detection were analyzed and the expression levels of the corresponding chemokines and pro-inflammatory factors were calculated.
  • MOTOR was overexpressed in monocytes by lentiviral infection, and the effect of overexpression of MOTOR on cytokine release by monocytes under LPS stimulation was detected by qPCR. It was found that overexpression of MOTOR could promote the response of monocytes to LPS (see Figure 3).
  • HEK293T cells were co-transfected with vsvg, rev, gag and Flag-MOTOR plasmids; the nucleic acid sequence of the biological factor MOTOR encoded by the MOTOR plasmid was shown in SEQ NO.1.
  • MOTOR OE MOTOR overexpressing cells
  • MOTOR OE and control cells were stimulated with 1 ⁇ g/ml LPS. After 24 h, the cells were collected by centrifugation and RNA was extracted.
  • C57BL/6 wild-type mice male, 6-7 weeks, 20-29 g
  • the cecum was probed and slowly pulled out of the abdominal cavity.
  • the cecum was ligated twice in the middle of the ligated segment. After confirming that there was no obvious bleeding, the cecum was returned to the abdomen.
  • 1 ml of 0.9% normal saline was subcutaneously administered for anti-shock treatment.
  • the mice were allowed to eat and drink freely after the operation. The room temperature was kept constant.
  • Exosomes were diluted with PBS to contain 1 ⁇ 10 9 exosomes per 200 ⁇ l, and 200 ⁇ l of exosome solution was injected into mice via tail vein injection at 24h, 48h, and 72h of sepsis modeling.
  • FIG. 1 shows that MOTOR is a circRNA with coding ability
  • A Schematic diagram of MOTOR and first-generation sequencing of junction site
  • B RNA coding ability predicted by bioinformatics, CDR1as and HOTAIR are classic circRNAs and lncRNAs that have been proven not to encode proteins but to function in the form of RNA, and Actin is a clear protein-coding gene
  • C Western blot shows that Flag antibody can detect Flag-MOTOR
  • D Flag-MOTOR immunofluorescence is shown in the middle.
  • Figure 2 shows that knocking down MOTOR inhibits the expression of chemokines and antigen presentation-related molecules.
  • RNA-seq (A) and qPCR (B) show the expression of chemokines and cellular antigen presentation-related molecules after knocking down MOTOR;
  • Figure 3 shows that overexpression of MOTOR promotes the analysis of chemokines in monocytes under LPS stimulation.
  • Figure 4 shows that exosomes containing MOTOR promote the survival time of septic mice.
  • a and B NTA and electron microscopy analysis showed that exosomes were removed by this method;
  • C Exosomes containing MOTOR were injected into mice by tail vein injection, and it was found that the survival time of septic mice was significantly increased.

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Abstract

一种hsa_circRNA基因MOTOR在制备用于治疗脓毒症的药物中的应用和药物。根据hsa_circRNA_056558的基因序列，通过分子克隆构建过表达hsa_circRNA_056558的载体，将该载体转染到细胞中，并通过超速离心收集该细胞产生的外泌体，外泌体中包含MOTOR。MOTOR可增强单核细胞的功能，用于脓毒症免疫调节治疗。

Description

MOTOR基因在制备用于治疗脓毒症的药物中的应用和药物 技术领域
本发明涉及circRNA编码小肽的基因技术领域，具体涉及一种MOTOR基因在制备用于治疗脓毒症的药物中的应用和药物。
背景技术
脓毒症(sepsis)是机体对感染的反应失调而导致危及生命的器官功能障碍，给人类健康带来了巨大威胁。我国多中心横断面研究显示：我国脓毒症患者约占ICU住院人数的20.6％，90天病死率高达35.5％。因此，深入研究脓毒症的发病机制和寻找新的治疗靶标分子，既是国家和社会重大疾病的防治需求，也是临床面临的巨大挑战。
免疫失衡是脓毒症发生发展的根本机制。虽然过度炎症反应导致的多器官损伤是脓毒症致病的重要环节，但脓毒症的高炎状态往往伴随着长期而持久的免疫抑制。单核细胞LPS耐受是脓毒症免疫抑制的关键特征。LPS耐受的单核细胞内化杀伤病原微生物的能力降低，在二次感染时不能做出有效的反应，使脓毒症患者二次感染的风险增加，而二次感染恰恰是脓毒症患者死亡的最主要原因。但是单核细胞发生LPS耐受的确切机制尚未阐明，目前缺乏调控单核细胞LPS耐受的分子药物，这是亟待重点攻关的核心问题。
发明内容
针对现有技术的不足，本发明提出了hsa_circRNA基因MOTOR在制备用于治疗脓毒症的药物中的应用，MOTOR逆转脓毒症单核细胞LPS耐受，延长小鼠的生存时间。
本发明的目的可以通过以下技术方案实现：
本申请第一方面提出MOTOR基因或其药用衍生物在制备治疗脓毒症的药物中的应用，所述MOTOR基因的核苷酸序列如SEQ NO.1所示。
进一步地，所述药物以适合于在使用者的外周血释放MOTOR基因或其药用衍生物的剂型使用。
本申请还提供一种药物，所述药物包括含有MOTOR基因的外泌体；所述MOTOR基因的核苷酸序列如SEQ NO.1所示。
将本发明的药物加入常规辅料，按照常规工艺，可以制成药学上可接受的各种剂型，如片剂、胶囊剂、口服液剂、锭剂、注射剂、软膏剂、颗粒剂或各种缓控释制剂等。
本发明药物的载体是药学领域中可得到的常见类型，包括：黏合剂、润滑剂、崩解剂、助溶剂、稀释剂、稳定剂、悬浮剂或基质等。
优选地，所述剂型为注射剂。
本发明的有益效果：
本申请的hsa_circRNA_056558的基因序列能够逆转脓毒症时单核细胞LPS耐受，促进细菌感染下单细细胞分泌的趋化因子、促炎细胞因子和抗原提呈基因的表达，调节脓毒症患者的固有免疫和适应性免疫功能，预防二次感染，提高生存时间。
附图说明
下面结合附图对本发明作进一步的说明。
图1为本申请的MOTOR定位于细胞质的小肽结构示意图；
图2为本申请的敲低MOTOR抑制炎症因子和趋化因子的表达图；
图3为本申请的过表达MOTOR促进炎症因子和趋化因子的表达图；
图4为本申请的Motor外泌体的粒径分析图。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例，都属于本发明保护的范围。
在本说明书的描述中，参考术语“一个实施例”、“示例”、“具体示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不一定指的是相同的实施例或示例。
本发明采用以下步骤进行验证MOTOR对脓毒症时单核细胞LPS耐受的影响，具体步骤参见附图1-4；
实施例1 MOTOR是具有编码能力的circRNA，能翻译出137aa的小肽
1.1通过PCR和Sanger鉴定MOTOR是一个circRNA，利用生物信息学预测MOTOR的编码能力，通过Western blot和免疫荧光技术验证MOTOR的编码能力和亚细胞定位，发现MOTOR可翻译出定位于细胞质的小肽(见图1)。
1.1.1 PCR及Sanger测序实验
(1)RNA提取：
a)准备所需试剂与耗材：Trizol；氯仿；异丙醇；DEPC水；Rnase-free不同规格枪头；1.5mLRnase-freep EP管
b)操作具体如下：收集人单核细胞THP1置于EP管中，每个EP管加入1ml的Trizol后，用移液器反复吹打至看不到细胞沉淀；离心机预冷至4℃，每个EP管中加入0.2ml氯仿，涡旋15s，室温静置5min；随后设置离心条件为12000rcf， 离心15分钟；离心结束后分为三层，上层为无色水状RNA，中间层与下层为有机苯酚氯仿层，小心取出EP管，垂直吸取上清0.4ml，加入0.5ml的异丙醇，颠倒混匀十次，室温静置10分钟；设置离心条件为12000rcf，10分钟；随后弃去上清，每个EP管中加入DEPC水配置的75％的乙醇，颠倒混匀；设置离心条件为7500rcf，5分钟，取出弃上清；瞬离15秒，弃上清，保留RNA沉淀；随后加入DEPC水30ul溶解沉淀后使用分光光度计测RNA浓度，随后使用DEPC水调整浓度至1ng/ul。
(2)逆转录PCR：使用逆转录试剂盒(PrimeScriptTM RT reagent Kit with gDNA Eraser，TAKARA，RR047A)对500ng的RNA进行逆转录PCR。
(3)PCR：将上一步逆转录得到的cDNA使用Rnase-free水稀释三倍，按照下面的体系配置：
PCR引物序列如下：
将配置好PCR体系后使用Bio-Rad PCR仪进行反应，将PCR产物送上海生工进行Sanger测序(附图1)。
1.1.2Western Blot实验
(1)提蛋白实验：
a)准备所需试剂与耗材：RIPA蛋白裂解液；Cocktail蛋白酶抑制剂；灭菌后的1.5mlEP管
b)具体操作如下：将FLAG-MOTOR质粒转染到12孔板的细胞中，加入100μl的RIPA和Cocktail抑制剂，冰浴10min；随后设置离心条件为12000rpm，4℃，20min；小心吸取上清即为所需蛋白。加入5xloading buffer，100℃加热5min。
(2)免疫印记实验
a)制胶：使用12％PAGE凝胶快速制备试剂盒(Bio-rad；64484826)配制12％SDS-PAGE胶：
1.分别取3ml的Resolver A和Resolver B溶液，加入3μl TEMED和30μl 10％AP。震荡混匀，注入制胶玻璃板中，使液面和短玻璃板上沿之间的距离比梳齿长0.5cm即可；分别取1ml的Stack A和Resolver B溶液，加入2μl TEMED和10μl 10％AP。震荡混匀，注入制胶玻璃板中，插入梳齿；
2.等待30分钟后，上层胶凝固，拔去梳齿即可用于电泳。
b)电泳：
1.将凝固好的胶板放入电泳槽中，加入足量的1xRunning buffer；
2.将煮好的蛋白样品涡旋混匀，瞬离后分别在点样孔加入5μL分子量蛋白标准样品和15μL蛋白样品；
3.设置电泳条件：恒压200v，45分钟；
c)转膜：
1.根据目的条带剪取对应大小的PVDF膜，并用甲醇醒膜10分钟；
2.电泳结束后，使用启胶器撬开玻璃板，并切割下分离胶，置于转膜缓冲液中；
3.使用三明治夹心法：正极(白色)-海绵垫-三层滤纸-PVDF膜-分离胶-三层滤纸-海绵垫-负极(黑色)，全程操作在转膜液中操作，一层层轻轻滚动，去除气泡。
4.设置转膜条件：恒流300mA，60分钟，加如1x转膜液；由于转膜过程产热需将转膜槽置于低温环境。
d)封闭：
1.将膜置于提前配好的封闭液(2.5g牛血清蛋白+50ml 1xTBST)中，常温摇床封闭1小时。
f)免疫反应：
1.使用1xTBST缓缓洗去表面封闭液；
2.配制一抗(5μl FLAG一抗+10ml1xTBST)，加入孵育盒中，使膜可以在盒中随摇床轻轻晃动，4℃过夜。
3.回收一抗，使用1xTBST洗膜3次，每次7分钟后常温孵育二抗，时间为60分钟，回收二抗，使用1xTBST洗膜3次，每次7分钟。将膜浸于1xTBST溶液中进行下一步。
g)显色反应
1.打开Eblot超灵敏全自动发光成像分析系统；
2.按照发光液A液：B液＝1:1混合均匀，移液器均匀铺至膜上，曝光拍照。
结果表明，可以检测到Flag-MOTOR，说明MOTOR可编码蛋白质。
1.1.3免疫荧光染色
(1)一抗孵育
收取Flag-MOTOR THP1细胞，4％的PFA室温固定1h，用PBS漂洗3次，每次5min；用Blocking medium(1％胎牛血清+0.5％Tritonx-100)室温封闭1h后，加入用PBS按1：200稀释Flag抗体，置于4℃条件下孵育过夜。
(2)二抗孵育
使用PBS漂洗3次，每次5min；然后将对应的二抗用PBS按1：400稀释 (另外DAPI按1：1000稀释)，室温孵育1h后用PBS漂洗3次，每次5min，指甲油封片；用共聚焦显微镜进行图片的采集。
结果表明，Flag-MOTOR主要存在于细胞质当中。
实施例2敲低MOTOR抑制LPS刺激下单核细胞释放的趋化因子和促炎细胞因子
2.1通过体外培养人单核细胞THP1，订购特异性靶向MOTOR circRNA的siRNA，利用NEON电转系统将靶向MOTOR的siRNA和对照siRNA转染进入细胞，并在转染后24h加入1μg/ml LPS，加LPS处理后24h收细胞。通过二代测序和qPCR检测细胞因子和抗原提呈相关因子的表达，发现敲低MOTOR可显著抑制它们的表达(见图2)。
2.1.1二代测序和分析
(1)收集细胞，500rcf室温离心5min，弃培养基，细胞沉淀用PBS清洗一遍，加入1ml Trizol。
(2)含有细胞的Trizol样品放入液氮中速冻30s，放入干冰寄送给公司进行二代测序。
(3)待公司返回测序数据后，分析趋化因子和促炎因子的表达，利用FPKM和z-score算法进行归一化处理，做热图反映相关基因的表达差异。
2.1.2 qPCR
(1)配制qPCR体系如下：
进行qPCR上机检测后分析得到的CT值，计算相应的趋化因子和促炎因子的表达水平。
结果表明，在二代测序和qPCR检测中，利用siRNA敲低MOTOR都能显著降低趋化因子和促炎因子的表达水平。
实施例3过表达MOTOR促进LPS刺激下单核细胞释放的趋化因子和促炎细胞因子
利用慢病毒感染的方式在单核细胞中过表达MOTOR，利用qPCR检测LPS刺激下过表达MOTOR对单核细胞释放细胞因子的影响，发现过表达MOTOR可促进单核细胞对LPS的响应(见图3)。
3.1慢病毒包装与纯化
(1)将vsvg，rev，gag和Flag-MOTOR质粒共转染HEK293T细胞；其中MOTOR质粒编码生物因子MOTOR的核酸序列如SEQ NO.1所示。
(2)在转染48小时后，收取培养皿中的上清液，更换新鲜的含有1％FBS的DMEM培养液。96小时后，第二次收集上清。
(3)加入体积1/5的病毒浓缩液，混匀，4℃静置过夜。
(4)将加了病毒浓缩液的上清离心，条件为4℃，11000rcf，30分钟，弃上清，沉淀即为病毒颗粒。用1.5ml DMEM培养基重悬病毒，放-80℃冰箱保存。
3.2慢病毒感染
(1)取2x105THP1细胞放入12孔板中，加入500μl病毒液，放入细胞培养箱培养。
(2)感染一周后，加入嘌呤霉素筛选，经筛选后的细胞即为过表达MOTOR的细胞(MOTOR OE)。
3.3趋化因子和促炎细胞因子的检测
(1)用1μg/ml的LPS刺激MOTOR OE和对照细胞，24h后离心收集细胞，提取RNA.
(2)利用qPCR检测趋化因子和促炎细胞因子的表达
结果显示，过表达MOTOR可显著提高趋化因子和促炎细胞因子的表达。
实施例4含MOTOR外泌体延长脓毒症小鼠的生存时间
收集过表达MOTOR的外泌体，通过尾静脉注射的方式将其注射到脓毒症小鼠体内，可显著延长脓毒症小鼠的生存时间。
4.1提取含MOTOR的外泌体
(1)收集293T和过表达MOTOR的293T细胞的上清，500g离心5min，弃沉淀，保留上清；2000g离心5min，弃沉淀，保留上清；12000g离心30min，弃沉淀，保留上清；200000g离心90min，弃上清，保留沉淀；用PBS重悬沉淀，200000g离心90min，弃上清，保留沉淀，用200ul PBS重悬。
(2)进行负染和电镜拍照
4.2将含MOTOR的外泌体及对照外泌体注射到脓毒症小鼠体内，观察小鼠的生存时间
4.2.1盲肠结扎穿刺法(Cecum ligation and puncture，CLP)建立脓毒症小鼠模型
(1)C57BL/6野生型小鼠(雄性，6-7w，20-29g)，麻醉动物，于腹正中线剑突下约1cm处剪开约0.5cm，探及盲肠并将其缓慢牵拉到腹腔外，结扎盲肠，于结扎段正中2次，确认无明显出血后将避开盲肠还纳于腹中，术后皮下0.9％生理盐水1ml抗休克治疗，术后自由进食水，保持室温恒定。
(2)用PBS将外泌体稀释到每200μl含1x109个外泌体，在脓毒症造模的第24h、48h和72h通过尾静脉注射的方式将200μl外泌体溶液注射到小鼠体内。
(3)以第一次注射外泌体的时间记为0h，每隔24h观察并记录小鼠的生存 情况。
结果显示，注射含MOTOR的外泌体可以显著延长脓毒症小鼠的生存时间。
图1表示MOTOR是一个具有编码能力的circRNA；A.MOTOR示意图和junction site的一代测序；B.生信预测的RNA编码能力，CDR1as和HOTAIR分别是经典的被证明不编码蛋白，而以RNA形式发挥功能的circRNA和lncRNA，Actin是明确的蛋白编码基因；C.western blot显示Flag抗体可以检测到Flag-MOTOR；D.中表明Flag-MOTOR免疫荧光。图2是敲低MOTOR抑制趋化因子和抗原提呈相关分子的表达RNA-seq(A)和qPCR(B)显示敲低MOTOR后趋化因子和细胞抗原提呈相关分子的表达；
图3中过表达MOTOR促进LPS刺激下单核细胞分析的趋化因子A.过表达MOTOR后，MOTOR的表达水平；B.为qRT-PCR检测过表达MOTOR后，LPS刺激下CCL2，CCL3和CCL4的mRNA水平。
图4中含MOTOR外泌体促进脓毒症小鼠生存时间A和B.NTA和电镜分析显示通过该方法提到了外泌体；C.利用尾静脉注射的方式将含有MOTOR的外泌体注射到小鼠体内，发现可以显著增肌脓毒症小鼠的生存时间。
以上显示和描述了本发明的基本原理、主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是说明本发明的原理，在不脱离本发明精神和范围的前提下，本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明范围内。

Claims (7)

  1. MOTOR基因或其药用衍生物在制备治疗脓毒症的药物中的应用，其特征在于，所述MOTOR基因的核苷酸序列如SEQ NO.1所示。
  2. 根据权利要求1所述的应用，其特征在于，所述药物以适合于在使用者的外周血释放MOTOR基因或其药用衍生物的剂型使用。
  3. 根据权利要求2所述的应用，其特征在于，所述剂型为片剂、胶囊剂、服液剂、锭剂、气雾剂、注射剂、软膏剂、颗粒剂。
  4. 根据权利要求3所述的应用，其特征在于，所述剂型为注射剂。
  5. 一种用于预防或治疗脓毒症的药物，其特征在于，所述药物包括含有MOTOR基因的外泌体；所述MOTOR基因的核苷酸序列如SEQ NO.1所示。
  6. 根据权利要求5所述的药物，所述药物的剂型为片剂、胶囊剂、服液剂、锭剂、气雾剂、注射剂、软膏剂、颗粒剂。
  7. 根据权利要求6所述的药物，其特征在于，所述剂型为注射剂。
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