1.成都中医药大学针灸推拿学院,四川省成都市温江区柳台大道1166号,611137
2.山西中医药大学第二临床学院
tyjilaixi@126.com
收稿:2024-04-09,
修回:2024-07-11,
纸质出版:2024-09-17
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朱涛,李洁,尹浩等.麦粒灸对溃疡性结肠炎模型小鼠结肠组织JAK2/STAT3信号通路的影响[J].中医杂志,2024,65(18):1925-1933.
ZHU Tao,LI Jie,YIN Hao,et al.Effect of Grain-sized Moxibustion on JAK2/ STAT3 Signaling Pathway in Colon Tissue of Ulcerative Colitis Mice[J].Journal of Traditional Chinese Medicine,2024,65(18):1925-1933.
朱涛,李洁,尹浩等.麦粒灸对溃疡性结肠炎模型小鼠结肠组织JAK2/STAT3信号通路的影响[J].中医杂志,2024,65(18):1925-1933. DOI: 10.13288/j.11-2166/r.2024.18.013.
ZHU Tao,LI Jie,YIN Hao,et al.Effect of Grain-sized Moxibustion on JAK2/ STAT3 Signaling Pathway in Colon Tissue of Ulcerative Colitis Mice[J].Journal of Traditional Chinese Medicine,2024,65(18):1925-1933. DOI: 10.13288/j.11-2166/r.2024.18.013.
目的
2
观察麦粒灸“中脘”“天枢”“上巨虚”穴对溃疡性结肠炎(UC)小鼠结肠组织的影响并探讨其可能作用机制。
方法
2
40只C57BL/6雄性小鼠随机分为空白组 、模型组、麦粒灸组、美沙拉嗪组,每组10只。除空白组外,其余各组小鼠采用自由饮用3 %葡聚糖硫酸钠盐溶液法制备UC小鼠模型。造模成功后麦粒灸组小鼠予麦粒灸“中脘”“天枢”“上巨虚”干预,每穴3壮,每壮时间约为30 s;美沙拉嗪组给予300 mg/kg美沙拉嗪肠溶片溶液灌胃;空白组、模型组仅抓取固定,不予任何干预。各组干预均每日1次,连续7天。观察各组小鼠一般情况并对小鼠疾病活动指数(DAI)进行评分;检测结肠长度、肠重指数及结肠黏膜损伤评分;ELISA法检测血清细胞间黏附分子1(ICAM-1)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量;HE染色法观察结肠组织病理学变化;实时荧光定量PCR法和免疫组化法分别检测小鼠结肠组织中非受体型酪氨酸蛋白激酶2(JAK2)、信号转导及转录激活蛋白3(STAT3)、细胞因子信号转导抑制因子3(SOCS3) mRNA及阳性表达;免疫荧光双标染色法检测小鼠结肠组织JAK2、SOCS3平均荧光强度表达。
结果
2
与空白组比较,模型组小鼠肛周不洁、大便不成形,结肠黏膜形态破坏;体质量降低,结肠长度缩短,DAI评分、肠重指数、结肠黏膜损伤及组织病理评分、血清ICAM-1、IL-6、TNF-α含量增加,
结肠组织JAK2、STAT3 mRNA及蛋白表达升高,JAK2平均荧光强度表达升高,SOCS3 mRNA、蛋白及平均荧光强度表达降低(
P
<0.01)。与模型组比较,麦粒灸组、美沙拉嗪组小鼠肛周得到改善,结肠黏膜形态较为完整;体质量、结肠长度增加,DAI评分、肠重指数、结肠黏膜损伤及组织病理评分、血清ICAM-1、IL-6、TNF-α含量减少,结肠组织JAK2、STAT3 mRNA及蛋白表达降低,JAK2平均荧光强度表达降低,SOCS3 mRNA、蛋白及平均荧光强度表达升高(
P
<0.05或
P
<0.01)。麦粒灸组与美沙拉嗪组各指标比较差异均无统计学意义(
P
>0.05)。
结论
2
麦粒灸“中脘”“天枢”“上巨虚”能够改善UC模型小鼠结肠黏膜的损伤,其机制可能与SOCS3调控JAK2/STAT3信号通路相关。
Objective
2
To explore the effect of grain-sized moxibustion at "Zhongwan (RN12)" "Tianshu (ST25)" and "Shangjuxu (ST37)" on the colon tissue of mice with ulcerative colitis (UC), and to analyze the potential mechanism.
Methods
2
Forty C57BL/6 male mice were randomly divided into blank group, model group, moxibustion group and mesalazine group, with 10 mice in each group. In all the groups except for the blank group, UC mouse model was prepared by freely drinking 3% sodium dextran sulfate solution. After successful modeling, the moxibustion group was treated with grain-sized moxibustion at Zhongwan, Tianshu and Shangjuxu , 3 cones per acupoint, 30 s of each cone. The mesalazine group was given 300 mg/kg of mesalazine enteric-coated tablet solution by gavage. The blank group and the model group were only fixed by grasping without any intervention. Each group was intervened once a day for 7 consecutive days. The general condition of mice in each group was observed, and the disease activity index (DAI) score was evaluated. The colon length, intestinal weight and colon mucosal injury score were detected. The contents of serum intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The pathological changes of colon tissue were observed by HE staining. The mRNA and protein expressions of non-receptor tyrosine protein kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) in colon tissue of mice were detected by real-time fluorescence quantitative PCR and immunohistochemistry, respectively. The mean fluorescence intensity of JAK2 and SOCS3 in colon tissue of mice was detected by immunofluorescence double staining.
Results
2
Compared to the blank group, the mice in the model group had unclean perianal area, unformed stool, destroyed colonic mucosal morphology, shortened body weight and colon length, increased DAI score, intestinal weight index, colonic mucosal injury and pathological score, serum ICAM-1, IL-6 and TNF-α levels, increased mRNA and protein expression of JAK2 and STAT3 in colon tissue, and decreased mRNA and protein expression of SOCS3 (
P
<0.01). Compared to those in the model group, the perianal area of mice in the moxibustion group and the mesalazine group was improved, and the colonic mucosal morphology was more complete; body weight and colon length increased, while DAI score, intestinal weight index, colonic mucosal injury and pathological score, serum ICAM-1, IL-6 and TNF-α levels decreased, with decreased mRNA and protein expression of JAK2 and STAT3 in colon tissue, and increased SOCS3 mRNA and protein expression (
P
<0.01 or
P
<0.05 ). There was no significant difference in any index between the moxibustion group and the mesalazine group.
Conclusion
2
Grain-sized moxibustion at "Zhongwan", "Tianshu" and "Shangjuxu" can improve the damage of colon mucosa in UC model mice, and the mechanism may be related to the key factors regulating JAK2 / STAT3 signaling pathway based on SOCS3.
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