1. Pancreatology. 2019 Jun;19(4):557-565. doi: 10....
बनाया गया: 25 नवंबर 2025
प्रश्न
- Pancreatology. 2019 Jun;19(4):557-565. doi: 10.1016/j.pan.2019.04.014. Epub 2019
Apr 30.
miR-148a suppresses autophagy by down-regulation of IL-6/STAT3 signaling in
cerulein-induced acute pancreatitis.
Miao B(1), Qi WJ(2), Zhang SW(1), Wang H(1), Wang C(1), Hu L(1), Huang GW(1), Li
SR(1), Wang H(1).
Author information:
(1)Department of Infectious Disease, Beijing Friendship Hospital, Beijing,
100010, China.
(2)Department of Infectious Disease, Beijing Friendship Hospital, Beijing,
100010, China. Electronic address: [email protected].
Acute pancreatitis (AP) is a type of sterile inflammation of the pancreas,
potentially leading to systemic inflammatory response syndrome or multiple organ
failure. An emerging evidence that dysfunction of miRNA expression may alter
pivotal physiological functions and lead to inflammation infiltration and
complication of multiple diseases, including AP. Here, the AP model was
successfully replicated using cerulein in聽vitro and in聽vivo. RT-qPCR was used to
detect low expression of miR-148a in AP. This study verified that IL-6 was a
direct target of miR-148a. Over-expression of miR-148a decreased the mRNA and
protein levels of IL-6 by RT-qPCR and Elisa. Moreover, over-expression of
miR-148a improved the pathological state of AP through H&E and MPO staining and
transmission electron microscopy. After over-expressing miR-148a, Western blot
and immunohistochemical method were used to confirm the reduction of
autophagosomes and autolysosomes, blockade of the levels of p-STAT3, LC3-II,
ATG7, ATG4c, Beclin1 and the increased p62 expression in AP. The expression of
LAMP-2 was not significantly different. In addition, IL-6 and AG490, the
IL-6/STAT3 signaling inhibitor, were used to verify the role of IL-6/STAT3
signaling in the regulation of miR-148a on autophagy in cerulein-induced AP
in聽vitro and in聽vivo. Taken together, our findings indicate that miR-148a
suppresses autophagy via regulating IL-6/STAT3 signaling in cerulein-induced AP
in聽vitro and in聽vivo. The miR-148a appears to be a promising candidate for the
gene therapy of AP.
Copyright 漏 2019 IAP and EPC. Published by Elsevier B.V. All rights reserved.
DOI: 10.1016/j.pan.2019.04.014
PMID: 31122822 [Indexed for MEDLINE]
- Drug Des Devel Ther. 2018 Oct 31;12:3665-3673. doi: 10.2147/DDDT.S163645.
eCollection 2018.
Efficacy and safety of Tongfu powder in acute pancreatitis patients with
gastrointestinal dysfunction: a clinical trial.
Miao B(1), Li FW(1), Zhang SW(1), Wang H(1), Qi WJ(1), Wang C(1).
Author information:
(1)Department of Infectious Disease, Beijing Friendship Hospital, Capital
Medical University, Beijing, People's Republic of China, [email protected].
OBJECTIVE: To evaluate the efficacy and safety of Tongfu powder for external
application on Sh茅nqu猫 (the umbilicus, hereafter, Tongfu powder) versus
mosapride in acute pancreatitis (AP) patients with gastrointestinal dysfunction.
METHODS: A total of 102 AP patients were diagnosed using the latest Atlanta
Criterion and recruited at the Department of Infectious Disease, Beijing
Friendship Hospital (Beijing, People's Republic of China) from August 2014 to
December 2016. Patients were randomized into the Tongfu powder group and
mosapride group using the random table. Information on scores (eg, the
gastrointestinal function score) on days 1 and 7 of hospitalization, biochemical
indicators (eg, interleukin [IL]-2 and IL-6), indicators for curative effects
(eg, first defecation time, bowel sound recovery time, hospitalization costs,
and duration) were collected and compared between the 2 groups.
RESULTS: The gastrointestinal function score decreased significantly after
treatment, and the changes were significantly different between the Tongfu
powder group and the mosapride group (P<0.05). Significantly shorter time to
first defecation and bowel sound recovery was observed in the Tongfu powder
group versus the mosapride group (P<0.05). The improvements of IL-2, IL-4,
intestinal fatty acid binding protein, motilin, and vasoactive intestinal
peptide in the Tongfu powder group were higher than those in the mosapride group
(P<0.05). There were no significant differences in hospital cost and length of
hospital stay between the 2 groups.
CONCLUSION: This study suggested that Tongfu powder for external application may
improve gastrointestinal function for AP patients compared with mosapride.
DOI: 10.2147/DDDT.S163645
PMCID: PMC6217005
PMID: 30464398 [Indexed for MEDLINE]
Conflict of interest statement: Disclosure The authors report no conflicts of
interest in this work.
- J Clin Microbiol. 2013 Dec;51(12):4249-51. doi: 10.1128/JCM.01709-13. Epub 2013
Sep 11.
Splenic abscess caused by Streptococcus gallolyticus subsp. pasteurianus as
presentation of a pancreatic cancer.
Su Y(1), Miao B, Wang H, Wang C, Zhang S.
Author information:
(1)Department of Infection and Critical Care Medicine, Beijing Friendship
Hospital, Capital University of Medical Sciences, Beijing, China.
Splenic abscesses caused by Streptococcus bovis are rarely reported in the
literature and are mainly seen in patients with endocarditis and associated
colonic neoplasia/carcinoma. We report the first case of splenic abscess caused
by Streptococcus gallolyticus subsp. pasteurianus (Streptococcus bovis biotype
II/2) as presentation of a pancreatic cancer.
DOI: 10.1128/JCM.01709-13
PMCID: PMC3838054
PMID: 24025909 [Indexed for MEDLINE]
- Inflammation. 2013 Aug;36(4):897-906. doi: 10.1007/s10753-013-9617-z.
Magnolol pretreatment prevents sepsis-induced intestinal dysmotility by
maintaining functional interstitial cells of Cajal.
Miao B(1), Zhang S, Wang H, Yang T, Zhou D, Wang BE.
Author information:
(1)Department of Infection, Beijing Friendship Hospital, Capital Medical
University, Beijing 100050, China.
The purpose of this study was to investigate the mechanism by which magnolol
treatment prevents lipopolysaccharide (LPS)-induced septic dysmotility in mice.
Sepsis was induced by intravenous tail vein injection of LPS (4 mg/kg body
weight). Animals were divided into three groups: the magnolol-treated septic
group, the placebo-treated septic group, and the control group. Intestinal
transit and circular smooth muscle contraction were measured 12 h after LPS
injection, and immunocytochemisty was performed to study the morphology of
interstitial cells of Cajal (ICCs). Stem cell factor (SCF) expression and c-kit
phosphorylation were determined by Western blot analysis, and the mRNA levels of
inducible NO synthase (iNOS) were determined by RT-PCR. Nitric oxide (NO)
content, superoxide dismutase (SOD) activity, and malondialdehyde (MDA)
concentration were detected using commercial kits. Intestinal transit and
muscular contractility were significantly lower in the LPS-treated group than in
the control group. Immunocytochemical experiments showed that the total number
of ICCs, and the total and average lengths of the ICC processes were
significantly decreased in the LPS-treated group compared with those in the
control group. In LPS-treated animals, magnolol pretreatment significantly
accelerated intestinal transit, increased circular muscle contraction, and
prevented ICC morphology changes. Phosphorylation of c-kit and expression of SCF
were significantly downregulated in LPS-treated animals compared with control
animals. Magnolol pretreatment prevented sepsis-induced decreases in c-kit
phosphorylation and SCF expression in LPS-treated animals. Magnolol pretreatment
prevented the sepsis-induced increase in NO concentration, iNOS expression, and
MDA concentration, and decrease in SOD activity in LPS-treated animals. Our
results suggest that magnolol treatment prevents sepsis-induced intestinal
dysmotility by regulating SCF/c-kit and NO signaling to maintain functional
ICCs.
DOI: 10.1007/s10753-013-9617-z
PMID: 23436158 [Indexed for MEDLINE]
基于苗彬目前发表的论文（好像近5年没有文章），现有课题（关于厚朴酚的基础课题：厚朴酚通过调控巨噬细胞代谢重编程途径抑制SAP肺损伤的机制研究和基于肺与大肠相表里理论研究厚朴酚对重症急性胰腺炎肺损伤的作用机理和TLR4/AKT与SCF/c-kit信号通路串话的分子机制（18年已结题）），设计博士研究计划书
विचार
उत्तर
下面给出一份围绕“厚朴酚‑巨噬细胞代谢重编程‑miR‑148a/IL‑6/STAT3‑自噬‑肺肠轴”主线、并承接您团队既往工作基础的博士研究计划书（可直接用于申请材料/开题报告的框架与内容要点）。文末附参考文献（限于您提供的4篇代表作）。如需，我也可以把它排版成可下载的Word版本。
一、课题题目（拟）
中文：
厚朴酚通过调控巨噬细胞代谢重编程与 miR‑148a–IL‑6/STAT3–自噬通路，重塑“肺‑肠轴”微环境以缓解重症急性胰腺炎相关肺损伤的机制研究
英文：
Magnolol Attenuates Severe Acute Pancreatitis–Associated Lung Injury by Reprogramming Macrophage Immunometabolism and Modulating the miR‑148a–IL‑6/STAT3–Autophagy Axis Along the Gut–Lung Axis
二、研究背景与科学问题
重症急性胰腺炎（SAP）常并发急性肺损伤/ARDS，是决定预后的关键并发症。其病理生理具有“肺‑肠轴”特征：肠屏障破坏与蠕动障碍 → 菌群和内毒素经门静脉/淋巴转位 → 全身炎症与肺泡‑毛细血管屏障受损；其中先天免疫细胞（尤其巨噬细胞）的代谢状态、炎症因子级联与自噬稳态互相牵制，塑造损伤结局。
本团队在相关通路与干预方面具备系统积累：
- 证实 miR‑148a 下调参与胰腺炎病程，且通过抑制 IL‑6/STAT3 信号降低自噬通量（小鼠 caerulein 模型，细胞与动物一致）（Pancreatology, 2019）[1]；
- 在脓毒症模型中，**厚朴酚（magnolol）**可维持肠道 SCF/c‑kit 信号与ICC功能、改善肠动力与氧化应激（Inflammation, 2013）[4]；
- 小型临床随机研究提示**“通腑散外敷神阙”可改善AP患者胃肠功能并有利于多项炎症/屏障相关指标（DDDT, 2018）[2]。
这些线索提示：厚朴酚可能一方面经巨噬细胞免疫代谢重编程抑制 IL‑6/STAT3‑驱动的异常自噬与炎症扩增，另一方面经SCF/c‑kit‑ICC‑肠动力**轴降低肠源性炎症负荷，从而改善 SAP‑相关肺损伤（SAP‑ALI）。
核心科学问题：
- 厚朴酚如何在体内重编程（reprogram）肺/胰相关巨噬细胞代谢（糖酵解↔氧化磷酸化/脂肪酸氧化），并由此改变其效应表型与炎症输出？
- miR‑148a–IL‑6/STAT3–自噬轴在 SAP‑ALI 中扮演何种“分子开关”，厚朴酚是否通过上调 miR‑148a发挥效应？
- TLR4/AKT炎症通路与SCF/c‑kit肠动力/ICC通路如何在“肺‑肠轴”上串话，共同决定 SAP‑ALI 的程度？能否形成从肠到肺的可干预信号链？
三、研究目标与假设
总体目标： 阐明厚朴酚通过巨噬细胞代谢‑炎症‑自噬联动与肺‑肠轴跨器官调控多通路协同，缓解 SAP‑ALI 的系统机制，并提出可转化的分子与功能性标志物。
中心假设： 厚朴酚通过抑制 TLR4/AKT‑mTOR/HIF‑1α 轴所驱动的促炎性糖酵解，促使巨噬细胞向炎症分辨/修复表型转化；同时上调 miR‑148a，抑制 IL‑6/STAT3‑异常自噬；在外周组织保全 SCF/c‑kit‑ICC 功能、恢复肠动力，减轻肠源性促炎负荷，最终改善 SAP‑ALI。
四、研究内容 / 具体目标（Aims）
Aim 1｜阐明厚朴酚对 SAP 中肺/胰相关巨噬细胞免疫代谢的重编程作用及其对肺损伤的决定性影响
- 模型与分组：
- 小鼠 SAP：① L‑arginine 高剂量诱导；② caerulein 超刺激+LPS 二次打击（增强系统炎症与ALI的可重复性）。
- 设对照、SAP、SAP+厚朴酚低/中/高剂量组；设置TLR4 抑制（TAK‑242）与AKT 抑制对照以验证通路依赖性。
- 关键指标与方法：
- 肺损伤表型： 病理评分、肺湿干比、BALF 蛋白/细胞计数、PaO₂/FiO₂。
- 巨噬细胞代谢： 支气管肺泡灌洗分离肺泡巨噬；胰腺巨噬细胞磁珠分选；Seahorse（OCR/ECAR）；^13C‑葡萄糖/棕榈酸示踪（目标：糖酵解与FAO流向）；线粒体膜电位/ROS。
- 信号通路： TLR4/AKT/mTOR/HIF‑1α 与代谢酶（HK2、PFKFB3、CPT1a）Western/qPCR。
- 功能验证： 代谢抑制剂（2‑DG 抑制糖酵解、Etomoxir 抑制FAO），及骨髓来源巨噬细胞（BMDM） adoptive transfer。
- 预期： 厚朴酚抑制促炎糖酵解、提升有氧代谢/FAO，降低促炎因子/NETs，改善ALI。
Aim 2｜揭示 miR‑148a–IL‑6/STAT3–自噬在 SAP‑ALI 中的关键作用，并解析厚朴酚是否经该轴发挥效应
- 测定： 胰腺、肺组织与血清外泌体中 miR‑148a表达；p‑STAT3、IL‑6、LC3‑II、p62、Beclin1、ATG7/ATG4c，自噬流（Bafilomycin A1/氯喹阻断）与TEM观察自噬体。
- 干预： 体内 miR‑148a agomir/antagomir；体外巨噬细胞/肺泡上皮细胞共培养；IL‑6 3′UTR双荧光素酶报告验证直接靶向；3‑MA/雷帕霉素双向调控自噬验证因果。
- 联动验证： 与Aim 1 的代谢读数联立分析（多变量模型），检验免疫代谢→IL‑6/STAT3→自噬的级联。
- 预期： SAP时 miR‑148a 下调、IL‑6/STAT3/自噬异常活化；厚朴酚上调 miR‑148a并下调该轴，减轻ALI（与[1]一致并外推至肺）。
Aim 3｜解析 TLR4/AKT 与 SCF/c‑kit在“肺‑肠轴”上的串话与厚朴酚的系统性效应；并开展小规模临床关联探索
- 动物层面（肠道）：肠动力学（活性炭推进/荧光标记）、肠通透性（FITC‑dextran）、ICC 形态（c‑kit 免疫荧光）、SCF 水平、肠系膜淋巴结/门静脉菌移位；16S rRNA 菌群分析（α/β多样性与功能预测）。
- 干预验证： **c‑kit 抑制剂（如 imatinib）**与 TLR4/AKT 抑制剂分别/合并干预，检验两通路在厚朴酚效应中的必要性。
- 临床探索（前瞻性观察队列/样本库）： 入组 SAP 患者，分层记录是否并发ALI；采集 血浆 miR‑148a、IL‑6/STAT3 活化标志、I‑FABP、D‑乳酸等；与呼吸学指标、SOFA/Marshall评分相关性建模；外敷通腑散作为可选伴随干预记录（安全性依从性与肠功能指标），以团队既往 RCT 结果为依据[2]。
- 预期： 厚朴酚通过改善 SCF/c‑kit‑ICC‑肠动力降低肠源性炎症输入；临床层面 miR‑148a/炎症‑自噬标志物与ALI严重度相关，支撑转化潜力。
五、技术路线（文字示意）
动物建模（双模型并行） → 厚朴酚干预 ± 通路抑制/遗传/代谢药理学 →
肺损伤综合表型评估 ↔ 巨噬细胞代谢‑表型‑信号（Aim1） ↔ miR‑148a/IL‑6/STAT3/自噬网络（Aim2） ↔
肠动力‑ICC‑屏障‑菌群与肺表型联动（Aim3） → 机制整合与关键节点验证 → 小型临床关联
六、关键方法与实验要点
- 动物与样本量： C57BL/6 小鼠（8–10 周龄，雌雄分层）；基于预实验肺损伤评分效应量（Cohen’s d≈0.8 的保守估计），每组 n=10–12 可达 80% 把握度（α=0.05）。精确样本量以预实验方差重估。
- 厚朴酚给药： 参考文献与预实验确定口服/腹腔给药剂量梯度（低‑中‑高），以有效与安全窗为先导。
- 代谢检测： Seahorse XF（OCR/ECAR）、^13C 同位素示踪、代谢酶活性/蛋白、线粒体功能（JC‑1/ATP/ROS）。
- 自噬通量： LC3‑II/LC3‑I 与 p62 结合 BafA1 阻断法判定通量，TEM 定性。
- 分子操作： miR‑148a agomir/antagomir 体内转染；双荧光素酶报告；BMDM 与 AEC 共培养。
- 菌群与屏障： 16S测序、生物信息学分析；FITC‑dextran；I‑FABP。
- 统计： 正态性/方差齐性检验；多因素方差/混合效应模型；多重比较校正；效应量与置信区间报告；预注册分析方案与盲法判读。
七、创新点
- 将巨噬细胞免疫代谢重编程置于 SAP‑ALI 的因果链上，连接到 miR‑148a–IL‑6/STAT3–自噬这一已被团队证实的关键轴[1]，形成代谢‑炎症‑自噬统一框架。
- 在“肺‑肠轴”尺度，证明 TLR4/AKT 与 SCF/c‑kit通路的跨器官串话，揭示厚朴酚的系统性机制。
- 从团队既有的**中医外治证据[2]出发，设置可转化标志物（血浆 miR‑148a、I‑FABP 等）**与临床结局的相关性桥梁。
八、预期结果与评价指标
- 主要终点：肺损伤综合评分、PaO₂/FiO₂、BALF 指标、存活率（必要时）。
- 机制终点：巨噬细胞代谢通路转换（OCR/ECAR、代谢基因）、miR‑148a/IL‑6/STAT3 活化状态、自噬通量、肠动力/ICC 完整性、菌群稳态指标。
- 转化终点：血浆 miR‑148a 与 SAP‑ALI 严重度/预后（AUC、OR、净重分类改善NRI）。
九、可行性与基础
- 文献与前期基础： 团队已完成 miR‑148a‑IL‑6/STAT3‑自噬轴在胰腺炎的验证[1]；厚朴酚改善ICC‑肠动力的动物证据[4]；通腑散改良AP患者胃肠功能的临床证据[2]。
- 平台： 具备分子与动物实验、代谢流分析、组织学与TEM、基础16S测序协作条件。
- 资源： 与感染/重症/消化内科联合，便于临床样本采集。
十、进度安排（4 年）
- 第1年： 模型与剂量学确立；Aim1 关键读数（代谢/肺表型）；预实验与样本量重估。
- 第2年： Aim2 完成（miR‑148a 操作与自噬因果）；与Aim1数据整合。
- 第3年： Aim3（肠动力‑ICC‑菌群‑肺联动；通路串话验证）；启动临床样本入组与基线化验。
- 第4年： 完成临床关联分析；机制整合与论文撰写/答辩/专利布局。
十一、风险与备选方案
- 厚朴酚对 miR‑148a 影响不显著： 扩大 miRNA 筛查（小RNA测序）寻找并行通路；或聚焦免疫代谢‑STAT3 直接调控。
- 自噬非关键： 同时评估细胞死亡方式（铁死亡/程序性坏死）并并行验证。
- 模型不稳定： 双模型并行（L‑Arg 与 Cer+LPS；必要时胆盐回流模型在大鼠验证）。
- 临床入组困难： 改为前瞻性生物样本库建设，以回顾性结局匹配统计。
十二、伦理与数据管理
- 动物实验遵循 3R 原则与盲法；
- 临床研究经伦理审批并取得书面知情同意；
- 数据按 FAIR 原则管理，关键原始数据与脚本入库保存与备份。
十三、预期成果与产出
- 论文 3–4 篇：
- 免疫代谢‑肺损伤机制 + 厚朴酚干预（基础研究）；
- miR‑148a–IL‑6/STAT3–自噬因果链（机制深入）；
- 肺‑肠轴串话与SCF/c‑kit‑ICC通路（系统层面）；
- miR‑148a 等血浆标志物与 SAP‑ALI 相关性（转化/临床）。
- 专利 1–2 项：厚朴酚给药/组合物或基于 miR‑148a 的诊断/伴随检测。
- 会议报告与青年基金储备申请。
十四、与既往工作的衔接
- 延展[1]： 把“miR‑148a 抑制 IL‑6/STAT3‑自噬”从胰腺局部扩展到肺与系统性层次，并嵌入巨噬细胞代谢逻辑。
- 呼应[4]： 将“厚朴酚‑SCF/c‑kit‑ICC‑肠动力”与“TLR4/AKT‑炎症‑代谢”建立跨器官串话。
- 转化[2]： 以前期 RCT 的胃肠功能改善为基础，加入客观分子标志物与ALI 结局桥接。
- 启示[3]： 关注肠源性细菌与系统炎症/肿瘤相关信号，为菌群‑宿主互作提供警示与拓展思路。
参考文献（限列代表作）
[1] Miao B, Qi WJ, Zhang SW, et al. miR‑148a suppresses autophagy by down‑regulation of IL‑6/STAT3 signaling in cerulein‑induced acute pancreatitis. Pancreatology. 2019;19(4):557‑565. doi:10.1016/j.pan.2019.04.014.
[2] Miao B, Li FW, Zhang SW, et al. Efficacy and safety of Tongfu powder in acute pancreatitis patients with gastrointestinal dysfunction: a clinical trial. Drug Des Devel Ther. 2018;12:3665‑3673. doi:10.2147/DDDT.S163645.
[3] Su Y, Miao B, Wang H, et al. Splenic abscess caused by Streptococcus gallolyticus subsp. pasteurianus as presentation of a pancreatic cancer. J Clin Microbiol. 2013;51(12):4249‑4251. doi:10.1128/JCM.01709‑13.
[4] Miao B, Zhang S, Wang H, et al. Magnolol pretreatment prevents sepsis‑induced intestinal dysmotility by maintaining functional interstitial cells of Cajal. Inflammation. 2013;36(4):897‑906. doi:10.1007/s10753‑013‑9617‑z.
可直接用于申请书的“亮点摘要”（150–200字）
本研究拟在团队“miR‑148a‑IL‑6/STAT3‑自噬轴”与“厚朴酚‑SCF/c‑kit‑ICC”两大前期成果基础上，构建“巨噬细胞免疫代谢‑炎症‑自噬‑肺肠轴”统一机制模型。通过双动物模型、代谢流与自噬通量测定、miRNA 操作及通路干预，阐明厚朴酚缓解 SAP‑ALI 的系统机制，并以血浆 miR‑148a 等作为转化标志物开展临床关联验证。研究将为 SAP‑ALI 提供可行的分子靶点与中药单体干预证据，兼具机制深度与转化前景。
如果需要，我可以把上述内容精炼成国家/省部级博士课题申请书版式（含摘要、立项依据、研究基础、研究方案、进度安排、经费概算表格等），或根据贵院模板进行逐项对齐与字数控制。