Tongfei Feng,1– 3,* Yanlin Zhou,1,3,* Bin Lv,1,3 Lijun Cai1,3

1Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310003, People’s Republic of China; 2The First Clinical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China; 3Key Laboratory of Digestive Pathophysiology of Zhejiang Province, The First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Chinese Medical University, Hubin Campus, Hangzhou, 310006, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Lijun Cai; Bin Lv, Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang, 310053, People’s Republic of China, Tel +86-571-86613545, Fax +86-571-87073763, Email 20051018@zcmu.edu.cn; lvbin@medmail.com.cn

Purpose: Through network pharmacology combined with molecular docking and in vivo validation, the study examines the unexplored molecular mechanisms of Tongxieyaofang (TXYF) in the treatment of irritable bowel syndrome (IBS). In particular, the potential pharmacological mechanism of TXYF alleviating IBS by regulating CHRM3 and intestinal barrier has not been studied.

Patients and Methods: LC-MS technique and TCMSP database were used in combination to identify the potential effective components and target sites of TXYF. Potential targets for IBS were obtained from Genecards and OMIM databases. PPI and cytoHub analysis for targets. Molecular docking was used to validate the binding energy of effective components with related targets and for visualization. GO and KEGG analysis were employed to identify target functions and signaling pathways. In the in vivo validation, wrap restraint stress-induced IBS model was employed to verify the change for cytoHub genes and CHRM3 expression. Furthermore, inflammatory changes of colon were observed by HE staining. The changes of Ach were verified by ELISA. IHC and WB validated CHRM3 and GNAQ/PLC/MLCK channel variations. AB-PAS test and WB test confirmed the protection of TXYF on gut barrier. The NF-κB/MLCK pathway was also verified.

Results: In TXYF decoction, LC-MS identified 559 chemical components, with 23 remaining effective components after screening in TCMSP. KEGG analysis indicated that calcium plays a crucial role in TXYF treated for IBS. Molecular docking validated the binding capacity of the effective components Naringenin and Nobiletin with cytoHub-gene and CHRM3. In vivo validation demonstrated that TXYF inhibits the activation of Ach and CHRM3 in IBS, and inhibits for the GNAQ/PLC/MLCK axis. Additionally, TXYF downregulates TNF-α, MMP9, and NF-κB/MLCK, while modulating goblet cell secretion to protect gut barrier.

Conclusion: TXYF inhibits Ach and CHRM3 expression, regulating the relaxation of intestinal smooth muscle via GNAQ/PLC/MLCK. Additionally, TXYF inhibits NF-κB/MLCK activated and goblet cell secretion to protect gut barrier.

Keywords: irritable bowel syndrome, tongxieyaofang decoction, Ach, CHRM3, GNAQ/PCL/MLCK, gut barrier

摘要：目的：本研究探讨了不同剂量的甲苯磺酸瑞马唑仑和丙泊酚联合瑞芬太尼麻醉对腹腔镜手术患者血流动力学和炎症反应的影响。方法：本研究纳入了90名腹腔镜手术患者，体重指数低于35 kg/m²，ASA II-III级。患者分为三组：低剂量瑞马唑仑组（A）、高剂量瑞马唑仑组（B）和丙泊酚组（C）。比较三组之间的血流动力学指标（如收缩压、舒张压、心率、平均动脉压）和炎症反应指标（如IL-6、SAA、CRP和PCT）的变化，以及舒芬太尼、瑞芬太尼、乌拉地尔和苯肾上腺素的剂量和拔管时间。结果：三组在拔管时间、舒芬太尼和瑞芬太尼的剂量、乌拉地尔和苯肾上腺素的使用率和平均剂量方面没有统计学上的显著差异。A组苯肾上腺素平均剂量低于B组和C组，差异有统计学意义。从T0到T2，各组之间的收缩压、舒张压、心率和平均动脉压没有统计学上的显著差异，IL-6、SAA、CRP或PCT水平也没有显著差异。结论：在腹腔镜手术中使用瑞马唑仑进行麻醉诱导和维持，可确保患者稳定的血流动力学和炎症反应。低剂量瑞马唑仑可能会降低手术期间苯肾上腺素等升压药的使用率和剂量。

摘要: 多药联合策略是提高肿瘤治疗效果的重要手段，也是临床肿瘤治疗的主流方案。以蟾皮和蟾酥为代表的中药活性成分具有高效、低毒、作用范围广、多靶点等优点，近年来已成为肿瘤联合治疗的理想靶标。蟾皮和蟾酥是源自中华大蟾蜍Bufo Bufo gargarizans Cantor或黑眶蟾蜍Bufo melanotictus Schneider的传统动物中药，在临床上作为抗肿瘤辅助药物治疗各种癌症和癌性疼痛显示出优异的疗效。其机制涉及诱导细胞凋亡，阻滞细胞周期，抑制细胞增殖、迁移和侵袭，抑制肿瘤血管生成，逆转肿瘤细胞的多药耐药性，调节多种信号通路和靶点。此外，基于纳米给药系统的多药联合策略可以实现蟾皮或蟾酥活性成分和其他抗肿瘤药物的精确负载，并携带药物克服生理和病理屏障，在肿瘤组织内完成高效富集，实现肿瘤细胞的靶向递药以及药物的控制释放，从而提高抗肿瘤疗效，并降低药物的毒副作用。本文综述了蟾皮、蟾酥联合化疗药物、靶向药物、镇痛药等药物的临床疗效和安全性；在细胞和动物模型上评估了蟾皮和蟾酥联合化疗、靶向治疗、放疗或热疗、中药、信号通路抑制剂等疗法的效果和机制；总结了蟾皮和蟾酥有效成分与化疗药物、小分子靶向药物、单克隆抗体、中药有效成分和光动力、光热治疗药物的共给药策略，为临床蟾皮和蟾酥的合理用药及新型给药系统的开发提供依据。

关键词: 蟾皮, 蟾酥, 联合抗肿瘤策略,共递送策略, 癌症, 癌痛

目的： 本研究利用RNA测序和网络药理学方法，探讨了五味子乙素（Schisandrin B）在缓解肾脏缺血再灌注损伤（RIRI）中的靶点、通路和机制。

方法： 通过采用苏木精-伊红（HE）染色和高碘酸-席夫（PAS）染色，以及测量血肌酐和尿素氮（BUN）水平，评估了五味子乙素对RIRI的影响。通过RNA-Seq分析了RIRI和Sch B+RIRI处理的小鼠模型中的基因表达差异。网络药理学技术用于寻找关键过程、靶标和通路。采用活性氧（ROS）、线粒体超氧化物和JC-1线粒体膜电位等测定方法评估了Sch B的抗氧化能力。使用分子对接验证了关键靶标与Sch B之间的相互作用，并通过qRT-PCR、Western blot和免疫荧光等方法确认了这些靶标和通路的表达。

结果： 在小鼠RIRI模型中，五味子乙素预处理显著减少了肾脏病理损伤、炎症反应和细胞凋亡。病理损伤评分从I/R组的4.33 ± 0.33降至Sch B处理组的2.17 ± 0.17和1.5 ± 0.22（p < 0.01）。肌酐和BUN水平也有所降低（从144.6 ± 21.05 µmol/L和53.51 ± 2.34 mg/dL降至50.44 ± 5.61 µmol/L和17.18 ± 0.96 mg/dL，p < 0.05）。转录组和网络药理学分析识别了四个关键靶标（AKT1、ALB、ACE、CCL5）和PI3K/Akt通路。药理学实验验证了Sch B调节这些靶标，减少了细胞凋亡和氧化应激，增强了肾功能恢复。

结论： 五味子乙素通过调节如AKT1、ALB、ACE和CCL5等关键靶标，并激活PI3K/AKT通路，减少氧化应激、炎症和细胞凋亡，从而在RIRI中改善肾功能恢复。

关键词： 肾脏缺血再灌注损伤；五味子乙素；转录组学；网络药理学；PI3K/Akt通路