Huang Qin, Dong Long-Cong, Zhang Rui-Bin, Zhu Ke-Li, Wu Qiao-Feng, Yu Shu-Guang
Acupuncture-Moxibustion and Tuina College, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.
Zhen Ci Yan Jiu. 2023 Feb 25;48(2):158-64. doi: 10.13702/j.1000-0607.20211284.
To observe the protective effect of electroacupuncture (EA) on the intestinal mucosal barrier and its relationship with the Notch/NF-κB signaling pathway in mice with ulcerative colitis (UC), so as to explore its mechanism of treating UC.
Male C57BL/6J mice were randomized into control, model and EA groups, with 6 mice in each group. The UC model was established by giving the mice with 2% Dextran Sulfate Sodium (DSS) for 7 days. EA (2 Hz/15 Hz, 0.2 mA) was applied at bilateral "Zusanli" (ST36) for 30 min, once a day for 7 days. The disease activity indexes ［DAI=(body weight index score+stool score+bleeding score)/3; 0-4 points］ of mice were calculated. The morphological changes of colonic tissues of mice in each group were observed by HE staining, and serum contents of TNF-α and IL-6 were detected by ELISA. Claudin-1 protein expression in colon tissue was detected by immunofluorescence, while the protein expression levels of Muc-2, Notch-1, MMP-9 in colon tissue were detected by immunohistochemistry. The real-time PCR method was used to detect the expression levels of Notch-1, Hes-1, NF-κB, TLR-4 and AKT mRNA in colon tissues.
After modeling, the DAI, serum TNF-α and IL-6 contents, Notch-1 and MMP-9 protein expression, the relative expression levels of Notch-1, Hes-1, NF-κB, TLR-4 and AKT mRNA in the colonic tissue were significantly increased (<0.001, <0.01) in the model group relevant to the control group. At the same time, Claudin-1 and Muc-2 protein expression were significantly reduced (<0.01). After the EA intervention, the increased DAI score, TNF-α and IL-6 contents, Notch-1 and MMP-9 protein expression, the relative expressions of Notch-1, Hes-1, NF-κB, TLR-4 and AKT mRNA, and the decreased Claudin-1 and Muc-2 protein expression were all reversed compared with the model group (<0.05, <0.01, <0.001). H.E. staining of the colonic tissue showed damage and infiltration of inflammatory cells in the model group, and those were significantly improved in the EA group.
EA can promote the recovery of intestinal mucosal barrier function and reduce inflammatory reaction in UC mice, which may be associated with its effects in inhibiting the excessive activation of the Notch/NF-κB signaling pathway.
观察电针（EA）对溃疡性结肠炎（UC）小鼠肠黏膜屏障的保护作用及其与Notch/NF-κB信号通路的关系，以探讨其治疗UC的机制。
将雄性C57BL/6J小鼠随机分为对照组、模型组和电针组，每组6只。通过给予小鼠2%硫酸葡聚糖钠（DSS）7天建立UC模型。在双侧“足三里”（ST36）施加电针（2Hz/15Hz，0.2mA）30分钟，每天1次，共7天。计算小鼠的疾病活动指数［DAI =（体重指数评分+粪便评分+出血评分）/3；0 - 4分］。通过HE染色观察各组小鼠结肠组织的形态变化，采用ELISA法检测血清TNF-α和IL-6含量。通过免疫荧光检测结肠组织中Claudin-1蛋白表达，采用免疫组化法检测结肠组织中Muc-2、Notch-1、MMP-9蛋白表达水平。采用实时荧光定量PCR法检测结肠组织中Notch-1、Hes-1、NF-κB、TLR-4和AKT mRNA的表达水平。
造模后，模型组小鼠的DAI、血清TNF-α和IL-6含量、Notch-1和MMP-9蛋白表达、结肠组织中Notch-1、Hes-1、NF-κB、TLR-4和AKT mRNA的相对表达水平均显著升高（<0.001，<0.01），与对照组相比差异有统计学意义。同时，Claudin-1和Muc-2蛋白表达显著降低（<0.01）。电针干预后，与模型组相比，升高的DAI评分、TNF-α和IL-6含量、Notch-1和MMP-9蛋白表达、Notch-1、Hes-1、NF-κB、TLR-4和AKT mRNA的相对表达以及降低的Claudin-1和Muc-2蛋白表达均得到逆转（<0.05，<0.01，<0.001）。结肠组织HE染色显示模型组有损伤及炎性细胞浸润，电针组明显改善。
电针可促进UC小鼠肠黏膜屏障功能恢复，减轻炎症反应，可能与其抑制Notch/NF-κB信号通路过度激活有关。
