Li Kui-Wu, Chu Hao-Ran, Ruan Jing-Ru, Wang Jiao-Jiao, Chen Jin-Yu, Zhu Shan-Shan, Zou Ling, Liao Lu-Min, Tong Ting-Ting
Second Clinical Medical College of Anhui University of CM, Hefei 230061, China.
Famous Doctor Clinic, Second Affiliated Hospital of Anhui University of CM/Anhui Acupuncture and Moxibustion Hospital, Hefei 230061; Clinical Research Institute of Acupuncture and Moxibustion of Anhui Academy of TCM, Hefei 230061; Anhui Clinical Medical Research Center of Acupuncture and Moxibustion, Hefei 230061.
Zhongguo Zhen Jiu. 2023 Feb 12;43(2):177-85. doi: 10.13703/j.0255-2930.20220108-k0007.
OBJECTIVE: To observe the effects of moxibustion on the stem cell factor (SCF)/tyrosine kinase receptor (c-kit) signaling pathway and immune function in rats with diarrhea irritable bowel syndrome (IBS-D), and to explore the mechanism of moxibustion for IBS-D. METHODS: Among 52 young rats born from 6 healthy pregnant SPF rats, 12 rats were randomly selected into the normal group, and the remaining 40 rats were treated with the three-factor combination method of maternal separation, acetic acid enema and chronic restraint stress to establish the IBS-D rat model. Thirty-six rats with successful IBS-D model were randomly divided into a model group, a moxibustion group, and a medication group, 12 rats in each group. The rats in the moxibustion group were treated with suspension moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37); the rats in the medication group were treated with intragastric administration of rifaximin suspension (150 mg/kg). All the treatments were given once a day for 7 consecutive days. The body mass, loose stool rate (LSR), the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were measured before acetic acid enema (35 days old), after modeling (45 days old), and after intervention (53 days old). After intervention (53 days old), HE staining was used to observe the morphology of colon tissue, and spleen and thymus coefficients were measured; ELISA method was used to detect serum inflammatory factors (tumor necrosis factor a [TNF-a], interleukin [IL]-10, IL-8), T-lymphocyte subsets (CD, CD, CD), value of CD/CD and immune globulin (IgA, IgG, IgM); real-time PCR method and Western blot method was used to detect the expression of SCF, c-kit mRNA and protein in colon tissue; immunofluorescence staining method were used to detect positive expression of SCF and c-kit. RESULTS: After intervention, compared with the normal group, in the model group, the body mass and the minimum volume threshold when AWR scored 3 were decreased (<0.01), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD, CD, CD/CD, IgA, IgG, IgM were increased (<0.01), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were decreased (<0.01), and the positive expression of SCF and c-kit was decreased (<0.01). Compared with the model group, in the moxibustion group and the medication group, the body mass and the minimum volume threshold when AWR scored 3 were increased (<0.01, <0.05), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD, CD, CD, CD/CD, IgA, IgG, IgM were decreased (<0.01, <0.05), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were increased (<0.01), and the positive expression of SCF and c-kit was increased (<0.01). Compared with the medication group, in the moxibustion group, the level of serum CD was decreased (<0.05), the value of CD/CD was increased (<0.01), and there was no significant difference in other indexes (>0.05). The expression of SCF and c-kit mRNA was positively correlated with the minimum volume threshold when AWR scored 3 and IL-10 (<0.01), and negatively correlated with remaining indexes (<0.01, <0.05). CONCLUSION: Moxibustion could reduce visceral hypersensitivity, improve symptoms of abdominal pain and diarrhea in IBS-D rats, and its mechanism may be related to up-regulation of the expression of SCF/c-kit signaling pathway and improvement of IBS-D immune function.
目的：观察艾灸对腹泻型肠易激综合征（IBS-D）大鼠干细胞因子（SCF）/酪氨酸激酶受体（c-kit）信号通路及免疫功能的影响，探讨艾灸治疗IBS-D的作用机制。 方法：从6只健康SPF级孕鼠所生的52只幼鼠中，随机选取12只作为正常组，其余40只采用母婴分离、醋酸灌肠和慢性束缚应激三因素联合法建立IBS-D大鼠模型。将造模成功的36只大鼠随机分为模型组、艾灸组和药物组，每组12只。艾灸组大鼠于“天枢”（ST25）、“上巨虚”（ST37）穴行悬灸；药物组大鼠灌胃给予利福昔明混悬液（150mg/kg）。所有处理均每日1次，连续7天。于醋酸灌肠前（35日龄）、造模后（45日龄）及干预后（53日龄）测量大鼠体质量、腹泻率（LSR）、腹部回撤反射（AWR）评分达3时的最小容量阈值。干预后（53日龄），采用HE染色观察结肠组织形态，测量脾脏和胸腺系数；采用ELISA法检测血清炎症因子（肿瘤坏死因子α[TNF-α]、白细胞介素[IL]-10、IL-8）、T淋巴细胞亚群（CD、CD、CD）、CD/CD值及免疫球蛋白（IgA、IgG、IgM）；采用实时荧光定量PCR法和Western blot法检测结肠组织中SCF、c-kit mRNA及蛋白表达；采用免疫荧光染色法检测SCF和c-kit阳性表达。 结果：干预后，与正常组比较，模型组大鼠体质量及AWR评分达3时的最小容量阈值降低（<0.01），LSR、脾脏和胸腺系数、血清TNF-α、IL-8、CD、CD、CD、CD/CD、IgA、IgG、IgM水平升高（<0.01），血清IL-10水平及结肠组织中SCF、c-kit蛋白和mRNA表达降低（<0.01），SCF和c-kit阳性表达降低（<0.01）。与模型组比较，艾灸组和药物组大鼠体质量及AWR评分达3时的最小容量阈值升高（<0.01，<0.05），LSR、脾脏和胸腺系数、血清TNF-α、IL-8、CD、CD、CD、CD/CD、IgA、IgG、IgM水平降低（<0.01，<0.05），血清IL-10水平及结肠组织中SCF、c-kit蛋白和mRNA表达升高（<0.01），SCF和c-kit阳性表达升高（<0.01）。与药物组比较，艾灸组大鼠血清CD水平降低（<0.05），CD/CD值升高（<0.01），其余指标差异无统计学意义（>0.0５）。SCF、c-kit mRNA表达与AWR评分达3时的最小容量阈值、IL-10呈正相关（<0.01），与其余指标呈负相关（<0.01，<0.05）。 结论：艾灸可降低IBS-D大鼠内脏高敏感性，改善腹痛、腹泻症状，其机制可能与上调SCF/c-kit信号通路表达、改善IBS-D免疫功能有关。
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