CN112826814A - 3-羟基丁酸或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用 - Google Patents
3-羟基丁酸或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用 Download PDFInfo
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Abstract
本发明属于生物医药领域，公开了3‑羟基丁酸或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用。本发明通过构建放射性肠道损伤动物模型验证，首次发现3‑羟基丁酸具有明显抗纤维化、减少炎症细胞浸润、促进肠道修复等作用，能有效治疗放射性肠道损伤，本发明显示3‑羟基丁酸及其衍生物及其组合物制成的药物、功能制剂等产品在医药学、功能食品领域具有广泛的应用前景。
Description
技术领域
本发明涉及生物医药领域，更具体地，涉及3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物在治疗放射性肠损伤上的应用。
背景技术
放疗是治疗盆腔恶性肿瘤的最有效手段之一，约35％-61％盆腔恶性肿瘤患者接受过盆腔放疗，据报道，中国仅2015年一年的恶性盆腔肿瘤新发病例人数估值超过50万。尽管放疗显著延长了患者的生存时间，但其对正常组织的物理性作用，会导致盆腹腔脏器的损伤。放射性肠损伤(radiation intestinal injury,RII)是指因盆腔恶性肿瘤如宫颈癌、子宫内膜癌、卵巢癌、前列腺癌、直肠癌、膀胱癌等接受放疗后引起的肠道损伤，范围涵盖小肠及大肠。根据起病时间及病程变化情况，可分为急性放射性肠损伤(acute intestinalinjury,ARII)与慢性放射性肠损伤(chronic radiation intestinal injury,CRII)，通常以3个月为界。超过75％的接受盆腔放疗的患者会发生ARII，约5％～20％的患者会发展为CRII。实际上，CRII的发病率极有可能被低估。报道称81％的盆腔放疗患者可出现消化道症状，但仅有55％的患者向专业医生求诊。
CRII的主要病理改变包括粘膜血管异常增生、粘膜下层纤维化、阻塞性动脉内膜炎、炎症细胞浸润等。CRII患者症状反复，迁延难愈，并呈现进行性加重的趋势。早期主要临床症状为便血、贫血、腹泻、腹部绞痛、肛门疼痛、里急后重、失禁、慢性吸收不良等，晚期易出现严重并发症如消化道出血、脓肿、穿孔、肠瘘、直肠狭窄、梗阻，重度贫血等，甚至引起死亡。临床诊治难度大，患者饱受煎熬，身心健康受到严重影响。据统计，约90％的CRII 患者可有永久性的排便习惯改变，50％患者表示其生活质量受到各种消化道症状的影响。此外，CRII患者可同时合并放射性小肠损伤、放射性结直肠损伤、放射性膀胱损伤、放射性盆腔损伤及原发肿瘤复发转移等情况，这给患者诊治方案的制定带来巨大的困难与挑战。目前，CRII缺乏根本防治手段，其治疗已成为困扰医务工作者及患者的一大难题。
目前，国内外对轻中度CRII主要采用药物治疗(如局部硫糖铝、类固醇、柳氮磺吡啶、甲硝唑、瑞巴派特和短链脂肪酸如丁酸钠等)，局部福尔马林治疗，内窥镜氩气凝固治疗(APC)，激光治疗和高压氧治疗等。药物治疗主要途径包括口服和保留灌肠。目前常规药物口服治疗的效果非常有限，保留灌肠对轻度CRII虽有一定疗效，但操作繁琐，患者依从性差。局部福尔马林治疗须特别谨慎，主要在手术室内进行，且有肛管溃疡、直肠狭窄、肛门失禁及肛门疼痛的风险。内窥镜治疗也可能带来肛门坠胀、疼痛、溃疡加重甚至直肠瘘的风险。采取以上几种方式治疗后，患者症状反复，或进行性加重，则易逐渐进展为重度CRII，需要手术治疗。
手术治疗主要有造口转流和病变肠管切除两种方式。单纯造口转流并不能切除受损组织，残留病变肠管仍导致顽固性症状，使患者面临持续并发症的风险。同时，长期造口易出现造口并发症，如造口脱垂，造口旁疝等，临床护理繁琐，部分患者也因难以忍受顽固性疼痛、穿孔等并发症而被迫选择病变肠管切除手术。病变肠管切除手术难度大，风险高，围手术期并发症常见。
在CRII的治疗决策中，综合临床症状与内镜表现，首先尽可能通过非手术治疗缓解主要症状，避免严重并发症的发生。而药物治疗因其治疗方式简便、患者依从性高而备受患者及医务工作者的青睐。然而，目前，CRII治疗药物非常少，且收效甚微，而CRII症状顽固，病变不可逆转，易逐渐进展出现严重并发症，从而需要行手术治疗。因此，目前临床上亟待开发出一种更为有效、安全、依从性高的CRII治疗药物或功能制剂。
发明内容
有鉴于此，本发明为克服上述现有技术所述的至少一种不足，提供3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物在治疗放射性肠损伤上的应用，解决手术治疗易产生并发症而CRII治疗药物非常少且收效甚微的技术问题。
为了解决上述存在的技术问题，本发明采用下述技术方案：
3-羟基丁酸(3-hydroxybutyric acid或3-HB)或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用。
3-羟基丁酸(3-HB)或其衍生物结构式如下：
式I和式II中的R1均为H、不同链长的直链或支链烷基(如C1-C20、C1-C10、C1-C6或C1-C3的直链或支链烷基)、不同链长的直链或支链烷氧基(如C1-C20、C1-C10、C1-C6或C1-C3 的直链或支链烷氧基)、环烷基(如C3-C6的环烷基)或芳基；
式I中的R2为H、不同链长的直链或支链的烷基(如C1-C20、C1-C10、C1-C6或C1-C3的直链或支链烷基)、环烷基(如C3-C6的环烷基)或芳基；
式II中的R2为无毒金属离子(如钠、钾或钙等)，式II中n值根据金属离子的价态确定。
所述3-羟基丁酸或其衍生物可以是D型或L型的，也可以是D型和L型的混合物。
具体可为：3-羟基丁酸(3-hydroxybutyric acid或3-HB)甲酯、3-羟基丁酸(3-HB)乙酯、3- 羟基丁酸(3-HB)或其盐(包括其钠盐、钾盐、钙盐等)、3-羟基己酸(3-hydroxyhexanoic acid或 3-HHx)甲酯、3-羟基己酸(3-HHx)乙酯、3-羟基己酸(3-HHx)或其盐(包括其钠盐、钾盐、钙盐等)。
本发明中所述3-羟基丁酸及其衍生物可通过各类聚羟基脂肪酸PHA的水解和醇解等多种方法得到，经过蒸馏纯化，通过GC分析确认纯度极高，没有双键等危害细胞生长的副产物 (Chen GQ,Wu Q.Microbial Production and Applications of ChiralHydroxyalkanoates.Appl Microbiol Biotechnol，67(2005)592-599)。
所述3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物可以被制成药品、保健品或食品功能制剂，优选制成口服制剂，更优选制成缓控释靶向制剂，用于本发明制备的制剂方法是本领域技术人员已知的。
本发明的3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物制成制剂可以通过口服、静脉内、肌内、动脉内、髓内、鞘内、心室内、透皮、皮下、腹膜内、鼻内、肠、局部、舌下或直肠等途径施用。
本发明的3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物制成制剂可以包括注射液、片剂、丸剂、粉剂、颗粒剂、胶囊、口服液、栓剂或膜剂等剂型，上述各种剂型的药物均可以按照药学领域的常规方法制备。
本发明提供3-HB或其衍生物或其组合物具有减轻过度纤维化作用，可治疗放射性肠损伤形成的肠壁过度纤维化，也可治疗涉及具有纤维化表现的病症。
本发明提供3-HB或其衍生物或其组合物具有减轻炎症反应的作用，可治疗放射性肠炎，也可治疗涉及肠道炎症反应表现的病症。
治疗放射性肠损伤主要可以从两方面进行：
①减轻照射后肠壁过度纤维化：本发明发现3-HB可有效减轻盆腔照射后小鼠肠壁纤维化。
②减轻照射后肠壁炎症细胞浸润：本发明发现3-HB可以改善盆腔照射后小鼠肠壁炎症细胞浸润，显著降低促炎症因子的分泌。
本发明与现有技术相比较有如下有益效果：本发明通过构建放射性肠道损伤动物模型验证，首次发现3-羟基丁酸具有明显抗过度纤维化、减轻肠壁炎症反应、促进肠道修复等作用，能有效治疗放射性肠道损伤，为放射性肠损伤的非手术治疗提供了一种新方向，具体给出了一种疗效更为显著的药物——3-羟基丁酸(3-hydroxybutyric acid或3-HB)或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物，其作为药物应用于治疗放射性肠损伤使得治疗CRII 更为有效、安全，依从性更高，具有广泛的应用前景。
附图说明
图1为C57BL/6J小鼠进行25Gy盆腔外照射示意图。
图2为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射后8周体重变化折线图。
图3为空白对照组、单纯照射组、高剂量照射+3-HB给药组小鼠照射后6个月生存曲线。
图4为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肛门往上1-2cm肠道肠镜检查截图及内镜下评分。
图5为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠道组织HE染色及MASSON染色结果及放射性损伤评分。
图6为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠道组织纤维化标志物fibronectin、T 1 collagen及T 3 collagen免疫组织化学染色及黏膜下层占肠壁全层厚度比例。
图7为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠壁巨噬细胞标志物F4/80免疫组织化学染色及肠壁浸润巨噬细胞计数。
图8为空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠壁组织炎症因子mRNA表达情况。
具体实施方式
下面通过具体实例对本发明进行说明，但本发明并不局限于此。
下述实施例中所使用的实验方法如无特殊说明，均为常规方法；下述实施例中所使用的试剂、生物材料等，如无特殊说明，均可从商业途径得到。
实施例1：3-羟基丁酸减轻RII小鼠模型的放射性损伤症状。
取80只健康雌性SPF级C57BL/6J小鼠，体重18-20g(购于北京维通利华实验动物技术有限公司)，随机均分为4组，每组20只小鼠：空白对照组、单纯照射组、低剂量3-HB给药组小鼠及高剂量3-HB给药组。其中空白对照组常规饲养，不给予任何其它处理；单纯照射组常规饲养，给予单次25Gy盆腔照射，照射范围为肛门往上边长1cm矩形区域(图1)；低剂量3-HB给药组及高剂量3-HB给药组在其饮水内分别给予75mg/kg、150mg/kg体重3- 羟基丁酸，实验全程小鼠自由取水，其余处理同单纯照射组。每组20只小鼠中，5只小鼠记录照射前一天及照射后8周每周体重，并于照射后第8周处死收取肛门往上1-2cm肠道组织用于后续实验；5只在照射后第8周进行内镜检查并评分；另外10只小鼠用于观察记录照射后半年的生存曲线。
统计与数据分析：体重百分比变化采用mean±SEM展示，采用Student’s-t test进行统计比较；小鼠放射性肠道损伤直肠镜检查半定量评分、肠道半定量放射性损伤评分采用 Mann-Whitney test进行统计比较；生存曲线采用log rank test进行统计比较；P值小于0.05视为有显著差异。
图2表示空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射后8周每周体重相较照射前一天体重变化百分比。
由图2可知，25Gy盆腔照射后小鼠体重较空白对照组小鼠显著降低，但是给予高剂量 3-HB可以有效抑制小鼠照射后体重下降，低剂量3-HB抑制小鼠体重下降结果不明显。
实验结果表明：药物显著改善25Gy盆腔照射引起的动物体重下降。
图3表示空白对照组、单纯照射组、高剂量3-HB给药组小鼠照射后半年内的生存曲线。
由图3可知，小鼠25Gy盆腔照射后2个月至6个月会出现死亡，而高剂量3-HB可显著延长小鼠25Gy盆腔照射后的生存时间。
实验结果表明：药物能够延长小鼠25Gy盆腔照射后的生存时间。
图4表示空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肛门往上1-2cm肠道内镜检查截图及内镜下评分。
小鼠放射性肠道损伤直肠镜检查半定量评分
表1小鼠放射性肠道损伤直肠镜检查半定量评分
表2小鼠放射性肠道损伤直肠镜检查半定量评分
注：该评分由一名内镜医生采用单盲方法独立评分。
由图4可知，25Gy盆腔照射后小鼠受照射肠段粘膜出现苍白水肿、糜烂、溃疡甚至穿孔，而不同剂量的药物处理，均可有效减轻上述内镜下症状，而高剂量处理组的效果较低剂量组更优。
实验结果表明：药物显著改善25Gy盆腔照射造成的内镜下肠道粘膜损伤。
结论：3-羟基丁酸可以显著减轻放射性肠道损伤小鼠模型的症状，主要表现为抑制照射后体重下降，延长生存时间，同时改善照射后肠粘膜内镜下损伤。
实施例2：3-羟基丁酸减轻RII小鼠模型肠道半定量病理评分。
(1)实验材料
HE染色：Harris苏木素，伊红染液。
MASSON染色：苏木精，无水乙醇，无水氯化铁，酸性品红，磷钼酸，冰醋酸，苯胺蓝浓盐酸。
MASSON染色主要试剂配制：
Weigent氏苏木素：A液：将1g苏木素加入100ml无水乙醇中微热溶解，室温保存；B液：将4ml 30％无水氯化铁溶液，1ml浓盐酸以及100ml蒸馏水混合，室温保存。使用时将A液B液等量混合即可，一般现混现用。
Masson丽春红酸性复红液：将丽春红0.7g，酸性复红0.3g，蒸馏水99ml，冰醋酸1ml混合溶解，室温保存。
1％冰醋酸水溶液：将冰醋酸1ml加入100ml水中，室温保存。
1％磷钼酸水溶液：将磷钼酸1g溶解于100ml水中，室温保存。
苯胺蓝水溶液：将苯胺蓝2g溶于98ml蒸馏水及2ml冰醋酸中，室温保存。
1％盐酸酒精：将1ml浓盐酸加到1000ml无水乙醇中，室温保存。
(2)实验方法
HE染色：
用于HE(伊红染色法hematoxylin-eosin staining)染色的小鼠肠道组织至于10％中性福尔马林固定液中24h，经流水冲洗、脱水、透明、石蜡包埋，制备4μm厚的连续石蜡切片。石蜡切片经常规脱蜡至水，苏木素室温染10min，自来水冲洗30-60s；1％盐酸酒精分化1s，自来水冲洗1min；伊红室温染5-10min；梯度酒精脱水；二甲苯透明；中性树胶封片。
MASSON染色：
用于MASSON染色的小鼠肠道组织至于10％中性福尔马林固定液中24h，经流水冲洗、脱水、透明、石蜡包埋，制备4μm厚的连续石蜡切片。石蜡切片经常规脱蜡至水，Weigent氏苏木素室温染5min，自来水冲洗3min；1％盐酸酒精分化1s，自来水冲洗1min，蒸馏水浸洗30s；酸性品红室温染5min，蒸馏水浸洗30s；滴加1％磷钼酸，显微镜下观察至肌纤维显红色，粘膜下层显淡粉红色，不水洗直接苯胺蓝室温染3min，1％冰醋酸浸洗30-60s；梯度酒精脱水；二甲苯透明；中性树胶封片。
肠道放射性损伤半定量评分表
表3肠道放射性损伤半定量评分表
注：该评分由一名病理科医生采用单盲方法独立评分，每项评分范围为0-1、0-2或0-3，每张病理切片各组织分层单项评分的总和为该切片的放射性损伤评分。
图5表示空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠道组织HE染色及MASSON染色结果及放射性损伤评分。
由图5可知不同剂量的药物都可以改善小鼠25Gy盆腔照射后受照射肠段的病理损伤程度，降低肠道放射性损伤半定量评分。
实验结论：药物可以在肠道病理层面上改善小鼠肠道放射性损伤。
实施例3：3-羟基丁酸减轻RII小鼠模型肠壁过度纤维化。
(1)实验材料
Fibronectin抗体，Type I collagen抗体，Type III collagen抗体，抗原修复液，DAB显色液 (中杉金桥)。
(2)实验方法
用于纤维标志物免疫组织化学染色的小鼠肠道组织置于10％中性福尔马林固定液中24h，经流水冲洗、脱水、透明、石蜡包埋，制备4μm厚的连续石蜡切片。石蜡切片经常规脱蜡至水，浸没于枸橼酸钠抗原修复液(10mM，pH6.0)高压复性，待上汽3min后缓慢冷却，冷却后PBS浸洗2次，5min/次；兔血清封闭液室温封闭15min；分别直接滴加稀释后的fibronectin、TypeI collagen、TypeIII collagen一抗(稀释度分别为1:400、1:50、1:100)，4℃ 12h，PBS浸洗2次，5min/次；滴加生物素化二抗，室温40min，PBS浸洗2次，5min/次；滴加三抗，室温40min，PBS浸洗2次，5min/次；DAB显色，镜下观察，适时终止，自来水冲洗5min；苏木素复染，室温30s，1％盐酸酒精分化1s，自来水冲洗5min；梯度酒精脱水；二甲苯透明；中性树胶封片。
奥林巴斯正置电子显微镜下观察小鼠肠道病理切片，在10倍目镜、10倍物镜下选取肠壁增厚最明显处肠段，使用电子显微镜自带软件测量肠壁全层厚度与粘膜下层厚度(尽量保持两条测量线平行且重叠)，同时计算粘膜下层厚度/肠壁全层厚度比值。
图6表示空白对照组、单纯照射组、低剂量3-HB给药组小鼠、高剂量3-HB给药组小鼠照射8周后肠道组织纤维化标志物fibronectin(Fibr)、Type I collagen(COL1)及TypeIII collagen (COL3)的免疫组织化学染色及黏膜下层占肠壁全层厚度比例。
由图6可知给药组小鼠肠壁纤维标志物着色较少，肠壁增厚较轻，粘膜下层厚度占肠壁全层厚度较小。
实验结论：3-羟基丁酸可以减轻25Gy盆腔照射小鼠肠壁黏膜下层过度纤维化。
实施例4：3-羟基丁酸减轻RII小鼠模型肠壁炎症反应。
(1)实验材料
免疫组织化学染色：
F4/80抗体，抗原修复液，DAB显色液(中杉金桥)。
荧光定量PCR：
引物，Trizol，SYBR Green染料，PCR逆转录试剂盒。
(2)实验方法
免疫组织化学染色：
操作流程基本与实施例3相同，F4/80一抗稀释度为1:50。
巨噬细胞浸润评分：
奥林巴斯正置电子显微镜下观察小鼠肠道病理切片，在10倍目镜、10倍物镜下选取肠壁增厚最明显处肠段，然后根据需要调整物镜倍数，根据实际所见进行巨噬细胞浸润评分。巨噬细胞浸润评分具体评分标准如下：1、粘膜固有层内散在巨噬细胞，粘膜下层及固有肌层无巨噬细胞；2、粘膜隐窝内少量巨噬细胞(<10个/隐窝)，粘膜下层及固有肌层无巨噬细胞；3、黏膜隐窝内大量巨噬细胞(>10个/隐窝)，粘膜下层及固有肌层内出现巨噬细胞；4、肠壁全层均有大量巨噬细胞。由一名病理科医生采用单盲方法独立评分。
荧光定量PCR：
1)动物组织RNA的提取
1.将前期准备好的小鼠肠道组织或人体肠道组织置于2ml EP管中，并加入1mlTRIzol；
2.向EP管中加入高压灭菌的直径2mm钢珠2个，直径5mm钢珠1个，封口胶密封管口，置于震荡仪中60Hz震荡至组织完全碎裂；
3.取出钢珠，室温放置5min；
4.每管EP管内加入0.2ml氯仿，，盖紧管盖，在振荡器上震荡10s；
5.室温静置至管内液体分层，4℃12000rpm离心15min；
6.小心取出EP管，此时样品分为三层：上层为含有RNA的无色水相层，中层为DNA、脂类和蛋白质等，下层为红色的苯酚-氯仿层。取上清透明溶液至新的EP管中，能吸取的水相层体积大约为400-500μl；
7.再向新的EP管中每管加入0.5ml异丙醇，振荡器震荡10s，室温放置20min后4℃12000rpm离心10min，小心吸去上清，留下白色沉淀；
8.向沉淀中加入1ml新鲜配制的75％酒精，颠倒5次混匀后12000rpm 4℃离心5min；
9.小心吸去上清，将管子倒扣在吸水纸上吸去管口溶液，管口敞开在通风橱内风干 10min；
10.用20μl DEPC水室温5min溶解RNA后测浓度。
2)RNA定量
使用Nanodrp 2000微量定量仪检测定量。打开机器盖子并在定量处滴入1μl DEPC水，按“blank”扣去容积的影响。然后用擦镜纸擦掉DEPC水，再滴入1μl待测RNA样品，执行“measure”操作，即可得到RNA的绝对浓度。一般所有样品的RNA浓度都会用DEPC 水调制1000μg/ml。
3)逆转录合成cDNA
1.参考TOYOBO公司RT-PCR试剂盒说明书，第一步按照如下反应体系进行加样：
表4
反应条件为37℃5min。
2.第二步反应体系加样：
表5
反应条件为37℃15min，98℃5min，4℃保存。
4)Real-Time PCR扩增
1.引物设计：
使用Primer premier 5.0软件设计相应的引物序列，采用序列相似性查询系统(BLAST) 分析引物的同源性，并仔细选择引物使其跨越外显子，并采用Oligo 6软件分析引物热动力学特征。所有引物均由广州艾基生物公司合成，实验所需具体引物序列如下：
表6
2.参照SYBR Green Master Mix试剂说明书，按照以下反应体系进行加样：
表7
3.PCR扩增反应
瞬时离心混匀，然后将反应管放入PCR仪中，按照以下参数设置进行Real-TimePCR 反应：95℃，7min；95℃，10s；60℃30s，循环45次；60℃30s；溶解曲线60℃-98℃；20℃10s。每组样品设置3个复孔，以保证实验结果的有效性和重复性。
图7显示高浓度3-羟基丁酸可显著减轻照射后肠壁巨噬细胞浸润。
图8显示3-羟基丁酸可降低CCL2、CCL7等促炎症趋化因子的转录。
结论：3-羟基丁酸可以减轻、抑制受照射肠段炎症反应。
Claims (8)
1.3-羟基丁酸或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用。
2.根据权利要求1所述的应用，其特征在于，所述3-羟基丁酸或其衍生物为D型或L型，或D型与L型的混合物。
3.根据权利要求2所述的应用，其特征在于，所述3-羟基丁酸或其衍生物包括3-羟基丁酸甲酯、3-羟基丁酸乙酯、3-羟基丁酸或其盐、3-羟基己酸甲酯、3-羟基己酸乙酯、或3-羟基己酸或其盐。
4.根据权利要求1所述的应用，其特征在于，所述3-羟基丁酸或其衍生物或可产生3-羟基丁酸或其衍生物的细菌组合物被制成药品、保健品或食品功能制剂。
5.根据权利要求4所述的应用，其特征在于，所述制剂为口服制剂。
6.根据权利要求5所述的应用，其特征在于，所述制剂为缓控释靶向制剂。
7.根据权利要求6所述的应用，其特征在于，所述制剂通过口服、静脉内、肌内、动脉内、髓内、鞘内、心室内、透皮、皮下、腹膜内、鼻内、肠、局部、舌下或直肠途径施用。
8.根据权利要求6所述的应用，其特征在于，所述制剂包括注射液、片剂、丸剂、粉剂、颗粒剂、胶囊、口服液、栓剂或膜剂。
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| WO2022134306A1 (zh) * | 2020-12-22 | 2022-06-30 | 中山大学附属第六医院 | 3-羟基丁酸或其衍生物或可产生该物质的细菌组合物在治疗放射性肠损伤上的应用 |
| CN113398109A (zh) * | 2021-06-22 | 2021-09-17 | 谈高 | 一种预防或治疗炎性肠病的酮体混合物及其应用 |
| CN117017967A (zh) * | 2023-08-31 | 2023-11-10 | 中山大学附属第六医院 | 三丁酸甘油酯及其衍生物在制备预防和/或治疗放疗引起的消化道并发症药物中的应用 |
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