CN117065005A - 一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法 - Google Patents
一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法 Download PDFInfo
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Abstract
本方法涉及抑郁症技术领域,且公开了一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,包括以下步骤:S1,制备免疫细胞;S2,制备免疫细胞外泌体多肽再生因子;S3,将免疫细胞外泌体多肽再生因子用混合物A进行乳化,得到混合物B。该利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,能够升高慢性不可预测应激模型大鼠脑组织中BDNF含量。在体药理结果显示,本发明的多肽能够减少小鼠悬尾实验中应激绝望模型小鼠的悬尾时间;也能减少小鼠强迫游泳实验中应激绝望模型小鼠的强迫游泳时间;同时,本发明的多肽能够减少慢性不可预测应激模型大鼠的觅食潜伏期,可作为临床抑郁症有效的候选治疗方法。
Description
技术领域
本方法涉及抑郁症技术领域,具体为一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法。
背景技术
抑郁症是现在最常见的一种心理疾病,以连续且长期的心情低落为主要的临床特征,是现代人心理疾病最重要的类型。
临床可见,心情低落和现实过得不开心,情绪长时间地低落消沉,从一开始的闷闷不乐到最后的悲痛欲绝,自卑、痛苦、悲观、厌世,感觉活着每一天都是在绝望地折磨自己,消极,逃避,最后甚至更有自杀倾向和行为。患者患有躯体化症状。胸闷,气短。
经检索,201580043454.X 用于治疗抑郁症的方法,涉及一种用于治疗抑郁症的方法,例如治疗难治性抑郁症的方法;其中所述治疗方案根据患者在单核苷酸多态性(SNP)rs4306882处的基因型进行调整。具体地讲,所述方法包括确定在rs4306882处是否存在G或T等位基因,以及施用氯胺酮或艾司氯胺酮的给药方案,其中所述给药方案经过调整,以向那些在rs4306882多态性位点处具有所述G等位基因(而不是T等位基因)的患者提供更高剂量和/或更高频率的所述氯胺酮或艾司氯胺酮。还公开了一种用于预测患有抑郁症的患者是否遗传上易于对阻断单胺神经递质的再摄取的抗抑郁药反应不良的方法,所述方法包括对所述患者进行基因分型以确定所述患者在SNP rs4306882处的基因型。
目前,抑郁症的治疗药物主要基于单胺类物质假说,如上述,201580043454.X 用于治疗抑郁症的方法,临床上常用的治疗方法包括靶向单胺类递质的药物治疗和电休克疗法等。但遗憾的是,这些药物都需要数周乃至数月才能发挥作用,并且有三分之一的患者对任何治疗方法都不反应。 因此, 研究新型有效的用于抑郁症治疗和预防的药物是治疗抑郁症的关键。
发明内容
针对现有技术的不足,本方法提供了一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,解决了上述的问题。
本方法提供如下技术方案:一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,包括以下步骤:
S1,制备免疫细胞;
S2,制备免疫细胞外泌体多肽再生因子;
S3,将免疫细胞外泌体多肽再生因子用混合物A进行乳化,得到混合物B。
优选的技术方案:所述步骤S1,包括以下步骤:
S101,用促进细胞贴壁材料对免疫细胞培养皿进行包被;
S102,依次将氨基酸、维生素、盐类、脂类或蛋白多肽中的一种或多种、细胞因子或抗体中的一种或多种营养物质加入到上述免疫细胞培养皿中,配制成免疫细胞培养基;
S103,对免疫细胞进行接种,将免疫细胞接种到上述免疫细胞培养基中,得到初步扩增的初代免疫细胞;
S104,将初步扩增的所述免疫细胞加入无血清免疫细胞诱导培养基进行第二阶段诱导扩增培养,得到诱导扩增的免疫细胞;
S105,诱导扩增的所述免疫细胞加入无血清免疫细胞激活培养基进行第三阶段激活扩增培养,得到激活扩增的免疫细胞。
优选的技术方案:所述步骤S2,包括以下步骤:
S201,将激活扩增的所述免疫细胞进行第四阶段培养,得到培养液;
S202,将培养液以密度梯度离心法,制备外泌体悬液;
S203,采用亲和层析柱进行蛋白纯化,收集洗脱液,得到所述组合免疫细胞外泌体多肽再生因子。
将培养液进行离心分离后,得上清液;然后采用MabSelectTMSuReTM亲和层析柱,使用5个柱体积平衡缓冲液进行平衡,将经步骤203初步纯化的样品加载到亲和层析柱上,用5个柱体积的淋洗缓冲液进行淋洗,使用5个柱体积的洗脱缓冲液进行洗脱,使用预放有中和液容器收集获得的目的蛋白洗脱物,中和后的目的蛋白溶液的pH为6.5~8.0。使用5个柱体积的再生液进行再生,使用3个柱体积的清洗液清洗层析柱,最后使用5个柱体积的平衡缓冲液平衡层析柱,最后使用保存液将层析柱保存;其中,平衡缓冲液:20mMTris-HCl,150mMNaCl,pH7.2;淋洗缓冲液:20mMTris-HCl,150mMNaCl,pH7.2;洗脱缓冲液:100mM柠檬酸缓冲液,0.1M蔗糖,pH3.4;中和液:1MTris-HCl,pH8.5;再生液:100mMHAc;清洗液:0.1MNaOH;保存液:20体积%乙醇。
优选的技术方案:所述步骤S3中的混合物A为弗氏完全佐剂与PBS缓冲液等体积混合得到。
优选的技术方案:所述步骤S101中的贴壁材料为胶原蛋白、明胶、多聚赖氨酸、纤粘连蛋白、层粘连蛋白、玻表粘连蛋白中的一种或多种。
优选的技术方案:所述步骤S102中的免疫细胞培养基还包括微量元素和其它小分子物质 ;其中所述微量元素包括氯化钴、亚硒酸钠、氯化镍、氯化锰、七钼酸铵、氯化铝、硫酸铬钾、硫酸铜、硝酸铁、硫酸亚铁或硫酸锌中的一种或多种 ;所述小分子包括 β- 巯基乙醇、还原型谷胱甘肽、D- 葡萄糖、牛磺酸或肝素钠中的一种或多种。
其通过采用氨基酸、维生素、盐类、脂类或蛋白多肽中的一种或多种,并和细胞因子或抗体中的一种或多种组成免疫细胞培养基,其能快速扩增不同类型的免疫细胞,让免疫细胞的扩增速度比常规培养基提高了 3-5 倍,延长了传代次数,而且能保持很好的免疫细胞功能。
而且本发明所述免疫细胞培养基所培养的免疫细胞能保持很好的免疫细胞功能,其能起到包括但不局限于抗抑郁、抗衰老的作用,一方面,经本发明所述免疫细胞培养基培养的免疫细胞像常规培养基所培养的免疫细胞一样,在其进行移植后对机体免疫系统具有修复作用 ;另一方面,经本发明所述免疫细胞培养基培养过的免疫细胞像常规培养基所培养的免疫细胞一样,经移植后可以恢复原有的功能。
优选的技术方案:所述步骤S103中的无血清免疫细胞诱导培养基基中包括以下成分:IL3000U/ml、IL101340U/ml、bFGF3ng/ml、BMP-50ug/ml、沙培林0.004KE/ml和层粘连蛋白10ug/ml。
优选的技术方案:所述步骤S101中的免疫细胞选自T细胞、NK细胞、细胞毒性T淋巴细胞、调节T细胞、记忆性T细胞、双特异性T细胞和CIK细胞中的任意一种。
优选的技术方案:在治疗抑郁症中的应用。
与现有技术相比,本方法提供了一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,具备以下有益效果:
该利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,实验证明,单次皮下注射给药序列表序列1或序列2所示多肽100μg/只,在大鼠强迫游泳实验中,与现有药物相比,可明显缩短大鼠在强迫游泳中的不同的时间,在慢性不可预见性应激实验中,可逆转慢性不可预见性应激所致的糖水偏爱值下降。本发明所提供的多肽,单次给药即可见效,作用持续时间长,对于给药后第50天测试的抑郁样行为仍然有治疗效果。本发明为预防和/或治疗抑郁症提供了一种新的药物及治疗方法,具有十分广阔的应用前景。
该利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,能够升高慢性不可预测应激模型大鼠脑组织中BDNF含量。在体药理结果显示,本发明的多肽能够减少小鼠悬尾实验中应激绝望模型小鼠的悬尾时间;也能减少小鼠强迫游泳实验中应激绝望模型小鼠的强迫游泳时间;同时,本发明的多肽能够减少慢性不可预测应激模型大鼠的觅食潜伏期,提高模型大鼠糖水实验中对糖水的摄取,与模型组相比,具有统计学差异。觅食潜伏期主要检测的是大鼠焦虑样行为,大鼠糖水偏好表明了大鼠的兴趣缺失倾向。由此可见,本发明多肽能改善急性和慢性抑郁症模型动物的抑郁症状。可作为临床抑郁症有效的候选治疗方法。
附图说明
图1为本方法的结构示意图;
图2为本方法的实施例一中的强迫游泳实验结果图;
图3为本方法的实施例一中的旷场实验结果图。
图中:1、溶媒对照组;2、多肽组;3、文拉法辛组。
具体实施方式
请参阅图1,一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,包括以下步骤:
S1,制备免疫细胞;
S2,制备免疫细胞外泌体多肽再生因子;
S3,将免疫细胞外泌体多肽再生因子用混合物A进行乳化,得到混合物B。
优选的技术方案:所述步骤S1,包括以下步骤:
S101,用促进细胞贴壁材料对免疫细胞培养皿进行包被;
S102,依次将氨基酸、维生素、盐类、脂类或蛋白多肽中的一种或多种、细胞因子或抗体中的一种或多种营养物质加入到上述免疫细胞培养皿中,配制成免疫细胞培养基;
S103,对免疫细胞进行接种,将免疫细胞接种到上述免疫细胞培养基中,得到初步扩增的初代免疫细胞;
S104,将初步扩增的所述免疫细胞加入无血清免疫细胞诱导培养基进行第二阶段诱导扩增培养,得到诱导扩增的免疫细胞;
S105,诱导扩增的所述免疫细胞加入无血清免疫细胞激活培养基进行第三阶段激活扩增培养,得到激活扩增的免疫细胞。
优选的技术方案:所述步骤S2,包括以下步骤:
S201,将激活扩增的所述免疫细胞进行第四阶段培养,得到培养液;
S202,将培养液以密度梯度离心法,制备外泌体悬液;
S203,采用亲和层析柱进行蛋白纯化,收集洗脱液,得到所述组合免疫细胞外泌体多肽再生因子。
将培养液进行离心分离后,得上清液;然后采用MabSelectTMSuReTM亲和层析柱,使用5个柱体积平衡缓冲液进行平衡,将经步骤203初步纯化的样品加载到亲和层析柱上,用5个柱体积的淋洗缓冲液进行淋洗,使用5个柱体积的洗脱缓冲液进行洗脱,使用预放有中和液容器收集获得的目的蛋白洗脱物,中和后的目的蛋白溶液的pH为6.5~8.0。使用5个柱体积的再生液进行再生,使用3个柱体积的清洗液清洗层析柱,最后使用5个柱体积的平衡缓冲液平衡层析柱,最后使用保存液将层析柱保存;其中,平衡缓冲液:20mMTris-HCl,150mMNaCl,pH7.2;淋洗缓冲液:20mMTris-HCl,150mMNaCl,pH7.2;洗脱缓冲液:100mM柠檬酸缓冲液,0.1M蔗糖,pH3.4;中和液:1MTris-HCl,pH8.5;再生液:100mMHAc;清洗液:0.1MNaOH;保存液:20体积%乙醇。
优选的技术方案:所述步骤S3中的混合物A为弗氏完全佐剂与PBS缓冲液等体积混合得到。
优选的技术方案:所述步骤S101中的贴壁材料为胶原蛋白、明胶、多聚赖氨酸、纤粘连蛋白、层粘连蛋白、玻表粘连蛋白中的一种或多种。
优选的技术方案:所述步骤S102中的免疫细胞培养基还包括微量元素和其它小分子物质 ;其中所述微量元素包括氯化钴、亚硒酸钠、氯化镍、氯化锰、七钼酸铵、氯化铝、硫酸铬钾、硫酸铜、硝酸铁、硫酸亚铁或硫酸锌中的一种或多种 ;所述小分子包括 β- 巯基乙醇、还原型谷胱甘肽、D- 葡萄糖、牛磺酸或肝素钠中的一种或多种。
其通过采用氨基酸、维生素、盐类、脂类或蛋白多肽中的一种或多种,并和细胞因子或抗体中的一种或多种组成免疫细胞培养基,其能快速扩增不同类型的免疫细胞,让免疫细胞的扩增速度比常规培养基提高了 3-5 倍,延长了传代次数,而且能保持很好的免疫细胞功能。
而且本发明所述免疫细胞培养基所培养的免疫细胞能保持很好的免疫细胞功能,其能起到包括但不局限于抗抑郁、抗衰老的作用,一方面,经本发明所述免疫细胞培养基培养的免疫细胞像常规培养基所培养的免疫细胞一样,在其进行移植后对机体免疫系统具有修复作用 ;另一方面,经本发明所述免疫细胞培养基培养过的免疫细胞像常规培养基所培养的免疫细胞一样,经移植后可以恢复原有的功能。
优选的技术方案:所述步骤S103中的无血清免疫细胞诱导培养基基中包括以下成分:IL3000U/ml、IL101340U/ml、bFGF3ng/ml、BMP-50ug/ml、沙培林0.004KE/ml和层粘连蛋白10ug/ml。
优选的技术方案:所述步骤S101中的免疫细胞选自T细胞、NK细胞、细胞毒性T淋巴细胞、调节T细胞、记忆性T细胞、双特异性T细胞和CIK细胞中的任意一种。
优选的技术方案:在治疗抑郁症中的应用。
实施例一,本发明多肽显著降低大鼠强迫游泳实验中的不动时间
实验方法
选取体重为220-240g的Sprague-Dawley(SD)雄性大鼠(购自北京维通利华实验动物技术有限公司),于温度22±2℃、 湿度50%±10%、12h/12h昼夜节律控制的动物房中,进行自由摄食饮水适应饲养4天后,将大鼠随机分为4组(分别记为溶媒对照组、多肽组、和文拉法辛组,每组10只大鼠),开始给药记为图1中的第0天),第8天进行5min旷场实验并记录爬格数,第9天进行5min强迫游泳测试并记录不动时间,实验流程如图1所示,实验重复三次,结果用平均值表示:
实施例所述给药的方式为单次皮下注射,各组间单次皮下注射的药物不同,具体如下:
溶媒对照组:每只大鼠注射药物溶媒(即等体积的PBS(0.1M,PH7.4)和弗氏完全佐剂 (购自Sigma-Aldrich公司 )的混合物0.5mL;
多肽组:每只大鼠注射乳剂A0.5mL(即每只100μg多肽);
文拉法辛组:每只大鼠按每千克体重注射溶液CK1.0mL(即注射文拉法辛40mg/kg);
本实施例所述旷场实验的具体方法如下:实验环境为长×宽×高=75cm×75cm×45cm的旷场,底部划分为25个面积相等的大方格。测试时将大鼠轻轻放入旷场实验区进行自由活动,进行5min自发活动的测定,记录大鼠的水平探索行为(即爬格数);
本实施例所述强迫游泳测试的具体方法如下:强迫游泳装置由透明有机玻璃制成(高24cm,直径15cm,水深17cm,水温24±2℃)。测试前一天,大鼠置于强迫游泳装置中适应15min;测试时,大鼠置于强迫游泳装置中,记录5min内大鼠漂浮不动时间。漂浮不动时间定义为大鼠微蜷躯体、呈漂浮状态的时间。强迫游泳实验为经典抗抑郁药物筛选模型,大多数抗抑郁药物可以有效降低大鼠在强迫游泳中的不动时间。
实验结果
采用SPSS软件进行数据分析, 实验数据均采用平均值±标准差(Mean±SEM)表示。 组间比较采用单因素方差分析(one-way ANOVA),Post hoc分析采用LSD检验,P<0.05时认为差异有统计学意义。
强迫游泳实验结果如图2所示,与溶媒对照组(71.5±18.5s)相比,多肽组(29±9s)大鼠强迫游泳不动时间显著降低(p<0.04),而文拉法辛组(65.04±6.27s)大鼠强迫游泳的不动时间没有降低(p>0.05)。
结果表明,多肽组单次给药即可在抑郁症的治疗和/或预防中产生作用,且效果优于传统抗抑郁药物文拉法辛。
旷场实验结果如图3所示,溶媒对照组(125±6)、多肽组(112±8)、和文拉法辛组(121.5±5)大鼠在旷场中的爬格数差异不显著,即多肽A91A96和多肽R91A96不影响大鼠的自发活动性。
实施例二:采用大鼠作为受试动物,取60只大鼠,雌雄各半,体重为180-220g,随机分为6组,高剂量组、中剂量组、低剂量组、阳性对照组、模型组、空白组,每组10只。建立大鼠慢性不可预测应激模型:应激内容为:束缚4h;湿垫料过夜;摇晃1h(160r/min);禁食12h;冰水游泳4℃,10min;饲养笼倾斜45°过夜;夹尾1min;拥挤过夜(8只/笼)。白天使用两种应激,夜晚使用一种应激,连续应激4周。最后一次刺激后给药。给药方案:治疗组皮下注射本发明多肽,剂量分别为20、10、5、2.5mg/Kg,对照组氟西汀,剂量为10mg/Kg,空白组给予生理盐水,连续给药7天。第7天给药后1h,大鼠断头处死,去脑组织,匀浆、离心,取上清,采用ELISA法(promega公司)检测BDNF含量;
下表为本发明多肽对慢性不可预测应激模型大鼠脑组织中BDNF含量的影响;
药物 | 剂量 | 动物数 | BDNF含量(μg/g脑组织) | |
模型组 | -- | -- | 10 | 0.05±0.1 |
阳光组 | 氟西汀 | 10mg/Kg | 10 | 0.63±0.38 |
高剂量组 | 本发明多肽 | 25mg/Kg | 10 | 0.91±0.38 |
中剂量组 | 本发明多肽 | 15mg/Kg | 10 | 0.80±0.21 |
低剂量组 | 本发明多肽 | 5mg/Kg | 10 | 0.43±0.51 |
本发明多肽能够升高慢性不可预测应激模型大鼠脑组织中BDNF含量,与模型组相比,具有统计学差异。
实施例三,糖水实验:
采用大鼠作为受试动物,取60只大鼠,雌雄各半,体重为180-220g,随机分为6组,高剂量组、中剂量组、低剂量组、阳性对照组、模型组、空白组,每组l0只。建立大鼠慢性不可预测应激模型:应激内容为:束缚4h;湿垫料过夜;摇晃1h(160r/min);禁食12h;冰水游泳4°C,10min;饲养笼倾斜45°过夜;夹尾1min;拥挤过夜(8只/笼)。白天使用两种应激,夜晚使用一种应激,连续应激4周。最后一次刺激后给药。给药方案:治疗组皮下注射本发明多肽,剂量分别为20、10、5、2.5mg/Kg,对照组氟西汀,剂量为10mg/Kg,空白组给予生理盐水,连续给药7天。第7天给药后1h,大鼠进行新奇觅食潜伏期实验和糖水偏好实验,以此评价本发明多肽对大鼠慢性不可预测应激模型的影响。新奇觅食潜伏期实验即:实验动物在进行24h禁食后,将动物置于一个长×宽×高为76.5cm×76.5cm×40cm的旷场中,旷场中间放有一定量的食物,动物从旷场的一角放入,观察动物在5min内成功觅食所花费的时间。同时试验后测量动物在5min内消耗的食物总重量,以此作为参考来判断实验动物饥饿程度是否一致。糖水偏好实验即:在糖水检测前用1%的糖水让动物适应48h,禁水4h后将动物置于放有两个相同水瓶的饲养笼中。这两只水瓶一只盛有1%的糖水,另一只装有自来水。记录1h内每只大鼠分别饮用糖水和自来水的体积。糖水偏好=饮用的糖水体积/(饮用的糖水体积+自来水总体积)×100%;
药物 | 剂量 | 动物数 | 觅食潜伏期(min) | 糖水偏好(%) | |
模型组 | -- | -- | 10 | 4.4±1.4 | 21.75±6.32 |
阳光组 | 氟西汀 | 10mg/Kg | 10 | 1.3±0.8 | 58±22 |
高剂量组 | 本发明多肽 | 25mg/Kg | 10 | 1.5±0.7 | 62.94±16.31 |
中剂量组 | 本发明多肽 | 15mg/Kg | 10 | 1.6±0.5 | 60.12±21.57 |
低剂量组 | 本发明多肽 | 5mg/Kg | 10 | 2.4±0.5 | 53.14±13.47 |
本发明多肽对大鼠慢性不可预测应激模型结果如表4显示:本发明多肽能够减少模型组大鼠的觅食潜伏期,提高慢性不可预测应激模型大鼠糖水实验中对糖水的摄取,与模型组相比,具有统计学差异。觅食潜伏期主要检测的是大鼠焦虑样行为,大鼠糖水偏好表明了大鼠的兴趣缺失倾向。由此可见,本发明多肽能改善慢性不可预测应激模型大鼠的抑郁症状。
Claims (9)
1.一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:包括以下步骤:
S1,制备免疫细胞;
S2,制备免疫细胞外泌体多肽再生因子;
S3,将免疫细胞外泌体多肽再生因子用混合物A进行乳化,得到混合物B。
2.根据权利要求1所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S1,包括以下步骤:
S101,用促进细胞贴壁材料对免疫细胞培养皿进行包被;
S102,依次将氨基酸、维生素、盐类、脂类或蛋白多肽中的一种或多种、细胞因子或抗体中的一种或多种营养物质加入到上述免疫细胞培养皿中,配制成免疫细胞培养基;
S103,对免疫细胞进行接种,将免疫细胞接种到上述免疫细胞培养基中,得到初步扩增的初代免疫细胞;
S104,将初步扩增的所述免疫细胞加入无血清免疫细胞诱导培养基进行第二阶段诱导扩增培养,得到诱导扩增的免疫细胞;
S105,诱导扩增的所述免疫细胞加入无血清免疫细胞激活培养基进行第三阶段激活扩增培养,得到激活扩增的免疫细胞。
3.根据权利要求2所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S2,包括以下步骤:
S201,将激活扩增的所述免疫细胞进行第四阶段培养,得到培养液;
S202,将培养液以密度梯度离心法,制备外泌体悬液;
S203,采用亲和层析柱进行蛋白纯化,收集洗脱液,得到所述组合免疫细胞外泌体多肽再生因子。
4.根据权利要求1所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S3中的混合物A为弗氏完全佐剂与PBS缓冲液等体积混合得到。
5.根据权利要求4所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S101中的贴壁材料为胶原蛋白、明胶、多聚赖氨酸、纤粘连蛋白、层粘连蛋白、玻表粘连蛋白中的一种或多种。
6.根据权利要求5所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S102中的免疫细胞培养基还包括微量元素和其它小分子物质 ;其中所述微量元素包括氯化钴、亚硒酸钠、氯化镍、氯化锰、七钼酸铵、氯化铝、硫酸铬钾、硫酸铜、硝酸铁、硫酸亚铁或硫酸锌中的一种或多种 ;所述小分子包括 β- 巯基乙醇、还原型谷胱甘肽、D- 葡萄糖、牛磺酸或肝素钠中的一种或多种。
7.根据权利要求1所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S103中的无血清免疫细胞诱导培养基基中包括以下成分:IL3000U/ml、IL101340U/ml、bFGF3ng/ml、BMP-50ug/ml、沙培林0.004KE/ml和层粘连蛋白10ug/ml。
8.根据权利要求1所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,其特征在于:所述步骤S101中的免疫细胞选自T细胞、NK细胞、细胞毒性T淋巴细胞、调节T细胞、记忆性T细胞、双特异性T细胞和CIK细胞中的任意一种。
9.根据权利要求1-8任一项所述的一种利用组合免疫细胞外泌体多肽因子治疗抑郁症的方法,在治疗抑郁症中的应用。
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