基于Th17/Treg免疫平衡探讨眼针治疗溃疡性结肠炎作用机制

Exploration of the Mechanism of Eye Acupuncture in the Treatment of Ulcerative Colitis Based on Th17/Treg Immune Balance

目的探讨眼针治疗溃疡性结肠炎的疗效作用机制。方法50只SD大鼠随机分为空白对照组、模型对照组、西药治疗组、眼针治疗组、针药联合组,每组10只。采用5%硫酸葡聚糖钠盐(dextran sulfate,DSS)水溶液自由饮用7 d建立溃疡性结肠炎(ulcerative colitis,UC)大鼠模型,每日灌胃、眼针(肺-大肠区)治疗1次,连续10 d。观察一般情况及每日疾病活动指数(disease activity index,DAI)评分,处死后行HE染色观察结肠组织病理学变化,酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法检测血清中白细胞介素(IL)-6、转化生长因子-β(transforming growth factor-β,TGF-β)、信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)含量,实时荧光定量PCR(q-PCR)检测结肠黏膜中叉状头转录因子(FoxP3)、维甲酸相关孤儿核受体γt(RORγt)、蛋白质酪氨酸激酶(JAK2)和STAT3 mRNA含量,Western blotting法检测结肠黏膜中FoxP3、RORγt、JAK2、STAT3和p-STAT3蛋白表达。采用SPSS 19.0进行统计学分析。结果与模型对照组相比,西药对照组、眼针治疗组和针药联合组DAI评分降低,病理损伤明显减轻,IL-6、TGF-β、RORγt、JAK2、STAT3、p-STAT3表达水平降低,FoxP3表达增加,差异有统计学意义(P<0.05)。西药对照组与眼针治疗组在IL-6、RORγt、JAK2、STAT3、p-STAT3表达方面有统计学差异(P<0.05)。结论眼针可以改善UC大鼠的临床症状,修复结肠黏膜损伤,其机制可能是通过抑制JAK2/STAT3通路以恢复Th17/Treg免疫平衡。

Objective To explore the mechanism of eye acupuncture in the treatment of ulcerative colitis(UC).Methods 50 SD rats were randomly divided into blank control group,model control group,Western medicine treatment group,eye acupuncture treatment group,acupuncture and medicine combination group,each group has 10 rats.The UC rat model was established by drinking 5%DSS water solution freely for 7 days.Gavage and eye acupuncture(lung-large intestine area)were given once a day for 10 consecutive days.Daily DAI score of rats,HE staining were used to observe the histopathological changes of colon,ELISA method was used to detect serum IL-6,TGF-β,STAT3 content,q-PCR to detect the colon tissue FoxP3,RORγt,JAK2 and STAT3 mRNA content,Western blotting was used to detect FoxP3,RORγt,JAK2,STAT3 and p-STAT3 protein expression in colon tissue.Results Compared with the model control group,the Western medicine control group,the eye acupuncture treatment group and the acupuncture combination group had lower DAI scores,significantly reduced pathological damage,and decreased expression levels of IL-6,TGF-β,RORγt,JAK2,STAT3 and p-STAT3,FoxP3 expression increased,and the difference was statistically significant(P<0.05).The Western medicine control group and the eye acupuncture treatment group had statistical differences in the expression of IL-6,RORγt,JAK2,STAT3and p-STAT3(P<0.05).Conclusion Eye acupuncture can improve the clinical symptoms of UC rats and repair the colonic mucosa injury.The mechanism may be to restore the Th17/Treg immune balance by inhibiting the JAK2/STAT3 pathway.