[1]吴珊,朱莹,张建伟,等.溃结宁膏穴位敷贴对溃疡性结肠炎大鼠氧化应激损伤的保护作用研究[J].西部中医药,2025,38(11):1-7.[doi:10.12174/j.issn.2096-9600.2025.11.01]
　WU Shan,ZHU Ying,ZHANG Jianwei,et al.Study on the Protective Effects of Kuijiening Ointment for Acupoint Application on Oxidative Stress Injury in Ulcerative Colitis Rats[J].Western Journal of Traditional Chinese Medicine,2025,38(11):1-7.[doi:10.12174/j.issn.2096-9600.2025.11.01]
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溃结宁膏穴位敷贴对溃疡性结肠炎大鼠氧化应激损伤的保护作用研究()

《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
38
期数:
2025年11期
页码:
1-7
栏目:
基础研究
出版日期:
2025-11-15

文章信息/Info

Title:
Study on the Protective Effects of Kuijiening Ointment for Acupoint Application on Oxidative Stress Injury in Ulcerative Colitis Rats
作者:
吴珊1, 朱莹2, 张建伟1, 王生华3, 张娜1, 王燚霈1
1.湖南中医药大学第二附属医院，湖南 长沙 410005
2.湖南中医药大学第一附属医院，湖南 长沙 410007
3.娄底市中医医院，湖南 娄底 417000
Author(s):
WU Shan1, ZHU Ying2, ZHANG Jianwei1, WANG Shenghua3, ZHANG Na1, WANG Yipei1
1.Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410005, China
2.First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, China
3.Loudi Hospital of Traditional Chinese Medicine, Loudi 417000, China
关键词:
结肠炎; 溃疡性; 溃结宁膏; 穴位敷贴; 信号通路; 线粒体融合蛋白1; 线粒体融合蛋白2; “虚、瘀”状态
Keywords:
colitis;  ulcerative; ointment; acupoint application; signaling pathway; mfn1; mfn2
分类号:
R285.5
DOI:
10.12174/j.issn.2096-9600.2025.11.01
文献标志码:
A
摘要:
目的以核因子E2相关因子2（nuclear factor-E2 related factor2，Nrf2）/血红素氧合酶-1（heme oxygenase-1，HO-1）信号通路调控线粒体融合蛋白-1（mitofusin-1，Mfn1）和线粒体融合蛋白-2（mitofusin-1，Mfn2）为研究切入点，探讨溃结宁膏穴位敷贴对“虚”“瘀”状态下溃疡性结肠炎（ulcerative colitis，UC）大鼠氧化应激损伤的保护作用及其潜在作用机制。 方法将51只SPF级雄性SD大鼠随机分为正常对照组（10只）与造模组（41只）。造模组大鼠采用腺嘌呤+冰番泻叶分阶段灌胃以及2，4，6-三硝基苯磺酸（2，4，6-trinitrobenzene sulfonic acid，TNBS）/乙醇法制备“虚”“瘀”状态下UC大鼠模型。待造模成功后，造模组大鼠再次随机分为模型组、非穴位敷贴组、穴位敷贴组、柳氮磺吡啶组并给予不同的干预方法。正常对照组、模型组给予无药物的巴布剂基质穴位敷贴；穴位敷贴组给予溃结宁膏穴位敷贴“足三里”“天枢”“肾俞”“膈俞”；非穴位敷贴组给予溃结宁膏穴位敷贴于臀部肌肉丰厚处；柳氮磺吡啶组给予无药物的巴布剂基质穴位敷贴加柳氮磺吡啶溶液灌胃，其余各组给予等量生理盐水灌胃，每日1次，连续干预28天。观察各组大鼠的死亡情况、宏观表征，评估结肠组织肉眼损伤评分与病理损伤评分；运用荧光定量PCR技术检测大鼠结肠组织Kelch样环氧氯丙烷相关蛋白1（Kelch-like ECH-associated protein-1，Keap1）、Nrf2、Mfnl、Mfn2 mRNA的表达水平；采用蛋白免疫印迹法（western blot，WB）检测大鼠结肠组织中keap1、Nrf2、HO-1蛋白的表达情况。 结果与正常对照组相比，模型组大鼠结肠组织肉眼损伤评分与病理损伤评分升高（P＜0.01），Keap1蛋白及mRNA表达升高（P＜0.01），Nrf2蛋白及mRNA、Mfnl mRNA、Mfn2 mRNA、HO-1蛋白表达明显降低（P＜0.05）；与模型组相比，穴位敷贴组、柳氮磺吡啶组大鼠结肠组织肉眼损伤评分与病理损伤评分降低（P＜0.01），Keap1蛋白及mRNA表达下调（P＜0.01），Nrf2蛋白及mRNA、Mfnl mRNA、Mfn2 mRNA、HO-1蛋白表达升高（P＜0.05），非穴位敷贴组上述指标变化无统计学意义；与非穴位敷贴组相比，穴位敷贴组大鼠结肠组织肉眼损伤评分与病理损伤评分降低（P＜0.01），Keap1蛋白及mRNA表达水平降低（P＜0.01），Nrf2蛋白及mRNA、Mfnl mRNA、Mfn2 mRNA、HO-1蛋白表达水平升高（P＜0.05）；穴位敷贴组和柳氮磺吡啶组各指标差异无统计学意义。 结论溃结宁膏穴位敷贴可通过激活Nrf2/HO-1信号通路，上调Mfnl、Mfn2 mRNA表达水平，改善线粒体损伤，从而发挥对UC大鼠氧化应激损伤的保护作用。
Abstract:
ObjectiveTo discuss the protective effects of Kuijiening ointment for acupoint application on oxidative stress injury in ulcerative colitis (UC) rats in "deficiency" and "stagnation" state, and its potential mechanism based on Nrf2/HO-1/Mfn1 and Mfn2. MethodsAll 51 SPF-grade male SD rats were randomized into normal control group (10 rats) and the modeling group (41 rats). The modeling group were prepared into UC rat models in "deficiency" and "stagnation" state by intragastric administration of adenine and iced senna leaves in stages and TNBS/ethanol method. After successfully modeling, the modeling group were randomly allocated to the model group, non acupoint application group, acupoint application group and sulfasalazine group, and they received different intervention methods. Normal control group and the model group were given medicine-free cataplasm matrix acupoint application, topical application of Kuijiening ointment at Zusanli (ST36), Tianshu (ST25), Shenshu (BL23) and Geshu (BL17) was administered to acupoint application group; topical application of Kuijiening ointment at well-muscled parts of the buttocks in non acupoint application group; sulfasalazine group received medicine-free cataplasm matrix acupoint application and gastric lavage of sulfasalazine solution, and the remaining groups were drenched with the equivalent amount of physiological saline, once each day, intervened for 28 days consecutively. To observe the mortality and macroscopic representation of the rats in different groups, to assess ocular trauma score (OTS) and pathological injury scores of colon tissue, fluorescent quantitative polymerase chain reaction (FQ-PCR) was adopted to measure the expressions of Keap1, Nrf2, Mfn1 and Mfn2 mRNA in colon tissue of the rats; WB was utilized to detect the levels of Keap1, Nrf2 and HO-1 protein in the colon tissue of the rats. ResultsCompared with normal control group, OTS and pathological injury scores of colon tissue were elevated in the model group (P<0.01), Keap1 protein and mRNA expressions increased (P<0.01), the levels of Nrf2 protein and mRNA, Mfn1 mRNA, Mfn2 mRNA and HO-1 protein remarkably reduced (P<0.05); compared with the model group, OTS and pathological injury scores of colon tissue decreased in acupoint application and sulfasalazine group (P<0.01), Keap1 protein and mRNA expressions down regulated (P<0.01), the expressions of Nrf2 protein and mRNA, Mfn1 mRNA, Mfn2 mRNA and HO-1 protein were raised (P<0.05), the differences had no statistical meaning in the changes of the above indexes in non acupoint application group; when acupoint application group was compared with non acupoint application group, OTS and pathological injury scores of colon tissue were lowered (P<0.01), the levels of Keap1 protein and mRNA were cut (P<0.01), the expressions of Nrf2 protein and mRNA, Mfn1 mRNA, Mfn2 mRNA and HO-1 protein up regulated (P<0.05); the difference showed no statistical meaning in the indexes between acupoint application group and sulfasalazine group. Conclusion Kuijiening ointment for acupoint application exert the protective effects against oxidative stress injury in UC rats by activating Nrf2/HO-1 signaling pathway, upregulating the levels of Mfn1 and Mfn2 mRNA and improving mitochondrial damage.

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备注/Memo

备注/Memo:
国家自然科学基金（81874466）；中医内科重大疾病防治研究与转化教育部重点实验室开放基金（ZYNK201705）；湖南省中医药科研计划项目（2020043）；湖南省教育厅科学研究项目（18C0401）。吴珊（1996—），女，硕士学位，医师。研究方向：消化系统疾病的中医药诊治。△；王燚霈（1985—），女，硕士学位，副主任医师。研究方向：消化系统疾病的中医药诊治。Email：wyp850815@126.com。
更新日期/Last Update: 2025-11-15