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上海中医药大学附属岳阳中西医结合医院,上海 200437
上海市针灸经络研究所,上海 200030
上海中医药大学,上海 201203
高倩倩,上海中医药大学硕士研究生。E-mail:gqq6600@163.com
✉周次利,副研究员。E-mail:zhoucili2010@126.com
收稿:2024-12-09,
网络首发:2025-12-26,
纸质出版:2026-04-12
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高倩倩, 冯芊茹, 彭洋, 等. 基于Trp/Kyn/AhR通路探讨艾灸干预克罗恩病肠纤维化小鼠的作用机制[J]. 中国针灸, 2026,46(4):589-600.
GAO Qianqian, FENG Qianru, PENG Yang, et al. Mechanism of moxibustion on intestinal fibrosis in Crohnʹs disease based on Trp/Kyn/AhR pathways in mice[J]. Chinese Acupuncture & Moxibustion, 2026, 46(4): 589-600.
高倩倩, 冯芊茹, 彭洋, 等. 基于Trp/Kyn/AhR通路探讨艾灸干预克罗恩病肠纤维化小鼠的作用机制[J]. 中国针灸, 2026,46(4):589-600. DOI: 10.13703/j.0255-2930.20241209-k0003.
GAO Qianqian, FENG Qianru, PENG Yang, et al. Mechanism of moxibustion on intestinal fibrosis in Crohnʹs disease based on Trp/Kyn/AhR pathways in mice[J]. Chinese Acupuncture & Moxibustion, 2026, 46(4): 589-600. DOI: 10.13703/j.0255-2930.20241209-k0003.
目的:
2
观察艾灸对克罗恩病(CD)肠纤维化小鼠结肠色氨酸(Trp)/犬尿氨酸(Kyn)/芳烃受体(AhR)通路的影响,探讨艾灸干预CD肠道炎症、肠纤维化的可能作用机制。
方法:
2
28只SPF级雄性C57BL/6小鼠,随机取8只作为正常组,剩余小鼠采用5%三硝基苯磺酸(TNBS)溶液灌肠建立CD肠纤维化模型。将造模成功的18只小鼠随机分为模型组、西药组和艾灸组,每组6只。艾灸组予温和灸“气海”和双侧“天枢”穴,每次10 min,每日1次,持续7 d;西药组予美沙拉嗪肠溶片混悬液0.5 mL灌胃,每日1次,持续7 d。观察各组小鼠的一般情况、疾病活动指数(DAI)评分、结肠长度、结肠大体宏观评分;HE、Masson染色观察小鼠结肠组织形态及肠纤维化情况,并计算胶原容积分数(CVF);ELISA法检测小鼠结肠白细胞介素(IL)-12、IL-23、Trp、Kyn含量;免疫荧光染色法观察小鼠结肠Collagen Ⅰ、Collagen Ⅲ、吲哚胺2
3-双加氧酶(IDO)阳性表达;Western blot法检测小鼠结肠Collagen Ⅰ、Collagen Ⅲ、IDO、AhR蛋白表达;实时荧光定量PCR法检测小鼠结肠IDO、AhR mRNA表达。
结果:
2
与正常组比较,模型组小鼠体质量降低、结肠长度缩短(
P
<
0.01),DAI评分、结肠大体宏观评分及组织学评分升高(
P
<
0.01),CVF升高(
P
<
0.01),结肠IL-12、IL-23、Trp、Kyn含量及Kyn/Trp比值升高(
P
<
0.01),结肠Collagen Ⅰ、Collagen Ⅲ蛋白表达升高(
P
<
0.01),IDO、AhR蛋白及mRNA表达升高(
P
<
0.01)。与模型组比较,艾灸组、西药组小鼠结肠长度增加(
P
<
0.01),DAI评分、结肠大体宏观评分、结肠组织学评分降低(
P
<
0.05,
P
<
0.01),CVF降低(
P
<
0.01),结肠IL-12、IL-23、Trp、Kyn含量及Kyn/Trp比值降低(
P
<
0.01,
P
<
0.05),结肠Collagen Ⅰ、Collagen Ⅲ蛋白表达降低(
P
<
0.05),IDO、AhR蛋白及mRNA表达降低(
P
<
0.01,
P
<
0.05)。与艾灸组比较,西药组小鼠结肠Trp、Kyn含量降低(
P
<
0.01)。
结论:
2
艾灸可改善TNBS诱导的CD小鼠的肠道炎症、肠纤维化,其机制可能与调节Trp/Kyn/AhR通路有关。
Objective
2
To observe the effect of moxibustion on tryptophan (Trp)/kynurenine (Kyn)/aryl hydrocarbon receptor (AhR) pathway in the colon of mice with intestinal fibrosis of Crohn's disease (CD)
and explore the mechanism of moxibustion on CD intestinal inflammation and intestinal fibrosis.
Methods
2
Among 28 male SPF C57BL/6 mice
8 mice were randomly selected as the normal group
and CD intestinal fibrosis models were prepared by enema with 5% trinitro-benzene sulfonic acid (TNBS) in the remaining mice as the modeling group. Eighteen successfully modeled mice were randomly divided into a model group
a western medication group
and a moxibustion group
6 mice in each one. In the moxibustion group
mild moxibustion was operated at "Qihai" (CV6) and bilateral "Tianshu" (ST25)
once daily
10 min each time and for consecutive 7 days. In the western medication group
the gavage with mesalazine enteric-coated tablet suspension
0.5 mL was administered
once a day and for consecutive 7 days. The general condition
score of disease activity index (DAI)
colon length
and colonic macroscopic score were observed in each group. HE and Masson staining were used to examine the morphology of the colon and intestinal fibrosis in mice
and the collagen volume fraction (CVF) was calculated. ELISA was adopted to detect the contents of interleukin (IL)-12
IL-23
Trp and Kyn in colon tissue. Using immunofluorescence assay
the positive expression of Collagen Ⅰ
Collagen Ⅲ and indolamine 2
3-dioxygenase (IDO) was observed. Using Western blot method
the protein expression of Collagen Ⅰ
Collagen Ⅲ
IDO and AhR in colon was detected. Using real-time quantitative PCR
the mRNA expression of IDO and AhR in colon was detected.
Results
2
Compared with the normal group
in the model group
the body mass was reduced and the colon length shortened (
P
<
0.01)
DAI score
the gross macroscopic score and histological score of colon increased (
P
<
0.01)
CVF was higher (
P
<
0.01)
the contents of IL-12
IL-23
Trp and Kyn and the ratio of Kyn to Trp were elevated (
P
<
0.01)
the protein expression of Collagen Ⅰ and Collagen Ⅲ increased (
P
<
0.01)
and the protein and mRNA expression of IDO and AhR were higher (
P
<
0.01). When compared with the model group
in the moxibustion group and the western medication group
the colon length was increased (
P
<
0.01)
DAI score
the gross macroscopic score and histological score of colon were reduced (
P
<
0.05
P
<
0.01)
CVF was lower (
P
<
0.01)
the contents of IL-12
IL-23
Trp and Kyn and the ratio of Kyn to Trp were reduced (
P
<
0.01
P
<
0.05)
the protein expression of Collagen Ⅰ and Collagen Ⅲ decreased (
P
<
0.05)
the protein and mRNA expression of IDO and AhR was declined (
P
<
0.01
P
<
0.05). In comparison with the moxibustion group
the contents of Trp and Kyn in the colon were lower in the western medication group (
P
<
0.01).
Conclusion
2
Moxibustion can attenuate intestinal inflammation and fibrosis induced by TNBS in CD mice
and its mechanism may be related to regulating Trp/Kyn/AhR pathway.
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