CN113717859A - 一种朱红硫磺菌及其应用 - Google Patents
一种朱红硫磺菌及其应用 Download PDFInfo
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- CN113717859A CN113717859A CN202110288535.3A CN202110288535A CN113717859A CN 113717859 A CN113717859 A CN 113717859A CN 202110288535 A CN202110288535 A CN 202110288535A CN 113717859 A CN113717859 A CN 113717859A
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Abstract
本发明涉及微生物技术领域，尤其是涉及一种朱红硫磺菌及其应用。本发明的菌株具有与SEQ ID NO:1所示的至少99％一致的18S rDNA‑ITS序列。本发明的菌株可以显著降低白细胞介素6(IL‑6)和肿瘤坏死因子(TNF‑α)的水平，进而治疗和预防白细胞介素6(IL‑6)和/或肿瘤坏死因子(TNF‑α)通路异常相关疾病，其起效快，绿色、安全、可靠。
Description
技术领域
本发明涉及微生物技术领域，尤其是涉及一种朱红硫磺菌及其应用。
背景技术
朱红硫磺菌，中文别名硫色多孔菌朱红色变种、红色硫黄菌、鸡冠菌等，属担子菌门(Basidiomycota)，伞菌亚纲(Agaricomycetes)，非褶菌目(Polyporales)，硫磺菌属(Laetiporus)，广泛分布于我国黑龙江、河北、四川、西藏、新疆、台湾等地区，是大型珍稀食药兼用真菌。子实体幼嫩至成熟时可食用，味道鲜美，营养丰富，老后呈干酪状，不宜食用，但可入药。性温，味甘，具有提高人体免疫力等多种功效，但由于其具体机制未明确，在现代疾病中的治疗适应症不明确。
白细胞介素-6(interleukin-6，IL-6)为由基质细胞、上皮细胞等多种细胞产生的一种多功能的细胞因子，在B淋巴细胞、T淋巴细胞、造血干细胞的增殖分化，炎症反应、自身免疫性疾病及某些肿瘤细胞的生长等过程中发挥重要作用。IL-6是一把双刃剑。其经典信号通路只在肝脏和免疫系统中表达，起到抑制炎症的作用；反式信号传导通路存在于几乎所有的细胞中，起到促进炎症的作用。血清IL-6水平作为炎症标志物在临床已广泛使用。IL-6上升的水平与炎性反应的严重程度、自身免疫性病的活动期、肿瘤的进展、器官移植排斥反应均密切相关。IL-6的升高与多种疾病相关，如(1)自身免疫性疾病，如类风湿性关节炎(rheumatoid arthritis,RA)、Costlemon病、银屑病等；(2)肿瘤，如多发性骨髓瘤(multiple myeloma,MM)、浆细胞瘤、慢性阻塞性肺炎、宫颈癌、霍奇金病等；(3)烧伤、术后、急性感染、器官移植排斥反应等。(4)代谢性疾病：如糖尿病、动脉粥样硬化。阻断IL-6信号通路的IL-6抗体已成为研发热点，如IL-6抗体托珠单抗，其适应症主要为类风湿性关节炎、幼年特发性关节炎等自身免疫病。其它已批或待批的IL-6抗体生物制剂药物适应症疾病为：巨细胞动脉炎(GCA)、Takayasu动脉炎(TA)、结节性动脉炎、结节性动脉炎等大血管炎，系统性红斑狼疮、肾小球肾炎、多发性骨髓瘤、难治性巩膜炎、CD19阳性的中枢神经系统白血病，CD19阳性的急性B淋巴细胞白血病，溃疡性结肠炎。
肿瘤坏死因子α(tumor necrosis factor-α，TNF-α)也是一种强效的细胞因子，其主要由巨噬细胞产生，参与人体诸多生理病理过程,包括自主免疫、癌症抑制和炎症发生等。TNF-α也是一柄双刃剑,一方面,它参与造血、先天免疫、细菌感染保护等有利于人体健康的生理过程；另一方面,它的异常调节也会引起类风湿性关节炎、心肌坏死和肿瘤形成等病理过程。其水平升高也被认为和很多疾病相关，目前上市的几种主要的TNF-α抗体包括阿达木单抗(adalimumab)修美乐、英夫利昔单抗(infliximab)类克等生物药，2014及2015年25种最畅销的药物中,前5名里就有3款是抗TNFα单抗。目前已批或待批的适应症疾病为：银屑病，炎症性肠病克罗恩病和溃疡性结肠炎、类风湿关节炎、强直性脊柱炎、干眼病、急性前葡萄膜炎、重度脓毒症。
目前，接受IL-6或TNF-α抗体治疗的患者发生感染、肝功能异常、血脂紊乱的风险有所升高，严重可导致患者住院或死亡。
发明内容
本发明的目的在于克服上述现有技术的不足之处而提供一种朱红硫磺菌及其应用，本发明的朱红硫磺菌可以显著降低白细胞介素6(IL-6)和/或肿瘤坏死因子(TNF-α)的水平，治疗和预防白细胞介素6(IL-6)和/或肿瘤坏死因子(TNF-α)通路异常相关疾病，起效快，绿色、安全、可靠，具有良好的经济效益和社会效益。
为实现上述目的，本发明采取的技术方案为：
本发明的第一方面提供了一种菌株，所述菌株具有与SEQ ID NO:1所示的至少99％一致的18S rDNA-ITS序列。
作为本发明所述菌株的优选实施方式，所述菌株为朱红硫磺菌(Laetiporussulphureus)ttys-997，所述朱红硫磺菌(Laetiporus sulphureus)ttys-997已于2020年12月3日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址为：北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.21084。
本发明发现一株可以降低白细胞介素6(IL-6)和肿瘤坏死因子(TNF-α)的水平的菌株，该菌株具有与SEQ ID NO:1所示的至少99％一致的18S rDNA-ITS序列，该序列长度为578bp，经过基因比对，该菌株被鉴定为Laetiporus(硫磺菌属)，为Laetiporus sulphureus(朱红硫磺菌)。其分类单元为Eukaryota；Fungi；Dikarya；Basidionmycota；Agaricomycotina；Agaricomycetes；Polyporales；Laetiporus。
本发明的朱红硫磺菌(Laetiporus sulphureus)ttys-997可以显著降低炎症反应指标白细胞介素6(IL-6)和肿瘤坏死因子(TNF-α)的水平，使得可以治疗和预防白细胞介素6(IL-6)和肿瘤坏死因子(TNF-α)通路异常相关疾病。
本发明的朱红硫磺菌(Laetiporus sulphureus)ttys-997在固体培养基上生长旺盛，菌丝浓密，25-28℃培养，菌落圆形，表面干燥，5d开始大量生成气生菌丝，并产生黄色色素。
本发明的第二方面提供了上述菌株在制备治疗或预防白细胞介素-6通路异常疾病的药物中的应用。
作为本发明所述应用的优选实施方式，所述疾病包括自身免疫性疾病、肿瘤和代谢性疾病。优选地，自身免疫性疾病包括银屑病、类风湿性关节炎、幼年特发性关节炎、巨细胞动脉炎、Takayasu动脉炎、结节性动脉炎、结节性动脉炎等大血管炎、系统性红斑狼疮、肾小球肾炎、多发性骨髓瘤、难治性巩膜炎、视神经脊髓炎，特异性皮炎，过敏性鼻炎，乳糜泻，哮喘，炎症性肠病、强直性脊柱炎、白塞氏病。肿瘤包括多发性骨髓瘤、浆细胞瘤、慢性阻塞性肺炎、宫颈癌、肺癌、乳腺癌、肝癌、淋巴瘤、CD19阳性的中枢神经系统白血病、CD19阳性的急性B淋巴细胞白血病。代谢性疾病包括糖尿病性黄斑水肿、糖尿病性周围神经损伤，动脉粥样硬化。本发明的菌株还可以治疗烧伤、术后、急性感染、器官移植排斥反应等疾病，如细胞因子释放综合征(CRS)。
本发明的第三方面提供了上述菌株在制备治疗或预防TNF-α通路异常疾病的药物中的应用。
作为本发明所述应用的优选实施方式，所述疾病包括银屑病，炎症性肠病、类风湿关节炎、强直性脊柱炎、干眼病、急性前葡萄膜炎、重度脓毒症中的一种或多种。
本发明的第四方面提供了一种药物组合物，包括如上述菌株及在药学上可接受的载体。
作为本发明所述药物组合物的优选实施方式，所述药物组合物为片剂、胶囊剂、口服剂或冻干粉剂。
作为本发明所述药物组合物的优选实施方式，所述药学上可接受的载体包括脱脂奶、乳糖、葡萄糖、蔗糖、山梨糖醇、甘露糖、海藻糖、淀粉、阿拉伯胶、磷酸钙、藻酸盐、明胶、硅酸钙、细结晶纤维素、聚乙烯吡咯烷酮、纤维素、水、糖浆、甲基纤维素、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁、矿物油中的一种或多种。
本发明的第五方面提供了一种用于治疗或辅助治疗炎症、自身免疫性疾病和肿瘤的食品或保健品，其特征在于，所述食品或保健品中含有上述菌株。
更优选的是，所述食品或保健品包括上述菌株、含有上述菌株的裂解物、含有上述菌株的培养上清液、含有上述菌株的代谢产物中的任意一种。
与现有技术相比，本发明具有以下有益效果：
本发明提供了一种朱红硫磺菌(Laetiporus sulphureus)ttys-997，其可以降低炎症反应指标IL-6与TNF-α水平，呈剂量效应关系，可以治疗IL-6与TNF-α通路异常相关疾病，具有起效快，绿色、安全、可靠的优点。
附图说明
图1为朱红硫磺菌(Laetiporus sulphureus)ttys-997扩增的18S rDNA-ITS序列基因比对结果图。
具体实施方式
为更好的说明本发明的目的、技术方案和优点，下面将结合附图和具体实施例对本发明作进一步说明。
实施例1、朱红硫磺菌(Laetiporus sulphureus)ttys-997的保藏信息
本发明的朱红硫磺菌(Laetiporus sulphureus)ttys-997已于2020年12月3日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址为：北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.21084。
实施例2、朱红硫磺菌(Laetiporus sulphureus)ttys-997的分离、鉴定和培养
从树桩上采摘获得菌株子实体，取一块子实体，无菌处理后置于PDA平板上，28度静置培养，14天后生长出丝状菌丝体。取一部分丝状菌丝体置于液体培养基中摇床震荡培养28天，有大量红色颗粒生成。将该红色颗粒离心收集后进行基因分析鉴定，该菌株具有18S rDNA-ITS片段(如SEQ ID NO:1所示)，该序列长度为578bp，经过基因比对，再结合子实体外观形态，该菌株被鉴定为Laetiporus(硫磺菌属)，为Laetiporus sulphureus(朱红硫磺菌)。其分类单元为Eukaryota；Fungi；Dikarya；Basidionmycota；Agaricomycotina；Agaricomycetes；Polyporales；Laetiporus。本发明将其命名为朱红硫磺菌(Laetiporussulphureus)ttys-997。
朱红硫磺菌(Laetiporus sulphureus)ttys-997在市面上常规的固体培养基上生长旺盛，菌丝浓密，25-28℃培养，菌落圆形，表面干燥，5d开始大量生成气生菌丝，并产生黄色色素。
实施例3、菌株对炎症模型鼠血清IL-6与TNF-α含量的影响
1.动物模型：以大鼠为动物模型，灌胃给予5％硫酸葡聚糖钠构建结肠炎模型。
2.实验动物：SD大鼠，雄性180-220g。(饲养环境：22±2℃，湿度为45～65％，12h昼夜交替)；自由进食与饮水。
3.实验方法：
1)造模：单笼饲养，除空白组外，其余5组用5％硫酸葡聚糖钠(0.75g/kg)连续灌胃10天，每日1次，空白组给予等量生理盐水。
2)分组及给药：
大鼠按照体重随机分为空白组、模型组、实验低、中、高剂量组(分别添加1mL/100g、2mL/100g和4mL/100g的朱红硫磺菌(Laetiporus sulphureus)ttys-997)、整肠生0.15g/kg，每组10只。从第1天灌胃硫酸葡聚糖钠开始，隔8小时后分别灌胃给药，共14天。
3)取材：
取材前一天，单笼饲养，观察粪便形态，用粪便潜血试剂盒测定并评分。大鼠最末次给药后禁食12小时，乙醚麻醉，下腔静脉取血，用于白细胞介素6(IL-6)、肿瘤坏死因子(TNF-α)测定。
4)检测指标
观察血清IL-6和TNF-α含量情况。
表1本发明的菌株对大鼠血清IL-6与TNF-α水平的影响(mean±SD)
| 组别 | 动物数 | IL-6(pg/mL) | TNF-α(pg/mL) |
| 对照组 | 10 | 21.86±9.73 | 28.23±15.23 |
| 模型组 | 10 | 68.72±24.09<sup>**</sup> | 57.25±21.83<sup>**</sup> |
| 整肠生0.15g/kg组 | 10 | 39.98±16.69<sup>##</sup> | 30.89±9.55<sup>##</sup> |
| 低剂量组 | 10 | 57.68±30.14 | 45.09±17.95 |
| 中剂量组 | 10 | 43.26±17.31<sup>##</sup> | 36.31±13.82<sup>##</sup> |
| 高剂量组 | 10 | 36.53±16.01<sup>##</sup> | 30.11±11.47<sup>##</sup> |
**P＜0.01与对照组相比；#P＜0.05，##P＜0.01与模型组相比；
见表1所示，与对照组相比，模型组的IL-6与TNF-α均显著升高；与模型组相比，整肠生0.15g/kg组IL-6与TNF-α均显著降低(P＜0.01)，实验中剂量组、高剂量组IL-6与TNF-α显著降低(P＜0.05；P＜0.01)且呈剂量效应关系。以上结果表明，本发明的朱红硫磺菌(Laetiporus sulphureus)ttys-997对降低硫酸葡聚糖钠致大鼠慢性结肠炎的血清IL-6与TNF-α水平有较好的作用，说明本发明的朱红硫磺菌(Laetiporus sulphureus)ttys-997可以治疗IL-6与TNF-α通路异常相关疾病(包括炎性、自身免疫性疾病和肿瘤)。该菌株治疗IL-6与TNF-α通路异常相关疾病起效快，绿色、安全、可靠，具有良好的经济效益和社会效益。
最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。
SEQUENCE LISTING
<110> 孙长春
<120> 一种朱红硫磺菌及其应用
<130> 2021.01.26
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 578
<212> DNA
<213> 人工合成
<400> 1
tgcggaagga tcattaacga acaacggggt tcgccccttg tccacaaaca cacccccgtg 60
cacgttgaag gcccggctcg ttgagtgggt gggcgaccgc ccaggattcg tagcctcgct 120
ttcttacaca aacttcagaa tgtacatcag aatgtcttac gcgtgtaacg catcgtaata 180
taactttcag caacggatct cttggctctc gcatcgatga agaacgcagc gaaatgcgat 240
aagtaatgtg aattgcagaa ttcagtgaat cattgaatct ttgaacgcac cttgcgctcc 300
ttggcattcc gaggagcatg cctgtttgag tgtcatgaaa ctctcaaccc ctgccatctt 360
tgcggatgag cgtcggttgg attttggagg ttgccggact cgttcggctc ctcttgaaag 420
catagtaaag cttggacctg accgaccggg tggacgtgat agaaagtcac cgtcgaccga 480
agggtccgtc gctgaacggt tcaagctttg tttgtcgtct tcggacgaac atctctgacc 540
tctgacctca aatcaggtag gactacccgc tgaactta 578
Claims (10)
1.一种菌株，其特征在于，所述菌株具有与SEQ ID NO:1所示的至少99％一致的18SrDNA-ITS序列。
2.如权利要求1所述的菌株，其特征在于，所述菌株为朱红硫磺菌(Laetiporussulphureus)，所述朱红硫磺菌(Laetiporus sulphureus)已于2020年12月3日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCC No.21084。
3.如权利要求1或2所述的菌株在制备治疗或预防白细胞介素-6通路异常疾病的药物中的应用。
4.如权利要求3所述的应用，其特征在于，所述疾病包括自身免疫性疾病、肿瘤和代谢性疾病。
5.如权利要求1或2所述的菌株在制备治疗或预防TNF-α通路异常疾病的药物中的应用。
6.如权利要求5所述的应用，其特征在于，所述疾病包括银屑病，炎症性肠病、类风湿关节炎、强直性脊柱炎、干眼病、急性前葡萄膜炎、重度脓毒症中的一种或多种。
7.一种药物组合物，其特征在于，包括如权利要求1或2所述的菌株及在药学上可接受的载体。
8.如权利要求7所述的药物组合物，其特征在于，所述药物组合物为片剂、胶囊剂、口服剂或冻干粉剂。
9.如权利要求7所述的药物组合物，其特征在于，所述药学上可接受的载体包括脱脂奶、乳糖、葡萄糖、蔗糖、山梨糖醇、甘露糖、海藻糖、淀粉、阿拉伯胶、磷酸钙、藻酸盐、明胶、硅酸钙、细结晶纤维素、聚乙烯吡咯烷酮、纤维素、水、糖浆、甲基纤维素、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁、矿物油中的一种或多种。
10.一种用于治疗或辅助治疗炎症、自身免疫性疾病和肿瘤的食品或保健品，其特征在于，所述食品或保健品中含有如权利要求1或2所述的菌株。
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| CN113667608B (zh) * | 2021-03-24 | 2023-08-22 | 孙长春 | 一株朱红硫磺菌及其在治疗消化道溃疡和结肠炎中的应用 |
| CN116077538A (zh) * | 2022-08-30 | 2023-05-09 | 广州图腾生命医学研究有限公司 | 朱红硫磺菌在制备治疗糖尿病足的药物、食品或保健品中的应用 |
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