清肠温中方联合粪菌移植对DSS诱导的溃疡性结肠炎大鼠肠黏膜的保护作用及其机制研究

Pro tective Effect and Mechanism of Qingchang Wenzhong Decoction Combined with Fecal Microbiota Transplantation on Intestinal Mucosa of Rats with Ulcerative Colitis Induced by DSS

目的观察清肠温中方联合粪菌移植对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)大鼠肠黏膜的保护作用以及作用机制。方法将48只SD大鼠适应性饲养7 d后随机分为供菌组、干预组,其中供菌组大鼠(8只)正常进食饮水,每天早上10点采集新鲜粪便,制作粪便滤液。干预组大鼠(40只)随机分为5组:空白组、模型组、中药组、粪菌移植组、联合组。空白组自由饮用去离子水,同时予去离子水(1 mL/100 g)灌胃,模型组、中药组、粪菌移植组、联合组给予4.5%DSS溶液自由饮用7 d制备UC模型,同时于下午2点分别给予去离子水(1 mL/100 g)、清肠温中方(1 mL/100 g)、粪便滤液(1 m L/100 g)、清肠温中方(1 m L/100 g)联合粪便滤液(1 mL/100 g)进行灌胃。各组干预治疗7 d,每日测量大鼠体质量,检测大鼠粪便潜血、记录粪便性状,计算大鼠疾病活动指数(DAI)。干预结束后麻醉大鼠,腹主动脉取血,取结肠组织,光镜下观察大鼠结肠病理损伤并进行结肠组织病理学(HS)评价,化学比色法检测髓过氧化物酶(MPO)的活性,ELISA检测血清白细胞介素-6(IL-6)、转化生长因子-β(TGF-β)的表达。结果与空白组相比,模型组大鼠毛色、活动度、粪便潜血、粪便性状等一般情况较差,DAI最大,病理损伤明显,HS评分最重,血清IL-6、TGF-β,结肠MPO的表达水平明显升高。经过干预,中药组、粪菌移植组、联合组大鼠上述指标均有不同程度的好转,并下调血清IL-6、TGF-β,结肠MPO的表达,差异均有统计学意义(P<0.05或P<0.01)。在改善粪便潜血方面,联合组显著优于中药组及粪菌移植组(P<0.05或P<0.01)。结论清肠温中方、粪菌移植以及两者联合应用均可明显缓解DSS诱导UC大鼠的症状,并可通过下调IL-6、TGF-β的表达而发挥作用,且联合应用效果在改善便血方面优于两者单独应用,中西医结合势必将成为未来UC的治疗方向。

Objective:To study the protective effects and mechanism of Qingchang Wenzhong Decoction combined with fecal microbiotal transplantation on intestinal mucosa of rats with UC induced by DSS,and to explore a new idea of integrated traditional Chinese and Western medicine in the diagnosis and treatment of UC.Methods:After one week of adaptive feeding,48 rats were randomly divided into the donor group and the intervention group.The donor group(8 rats)ate and drank water normally.Fresh feces were collected every morning at 10 o′clock to make fecal filtrate.40 rats in the intervention group were randomly divided into 5 groups:the blank group,the model group,the traditional Chinese medicine group,the transplantation group and the combination group.The blank group drank deionized water freely and was given deionized water(1 m L/100 g)by gavage.The other groups were given 4.5%DSS solution for 7 days to make UC model.At the same time,they were given corresponding drugs by gavage respectively at 2 pm.The rats′body weight,fecal occult blood,fecal characteristics and disease activity index(DAI)were measured daily for 7 days.After the intervention,blood was taken from the abdominal aorta and colon tissue was taken.HE stained paraffin sections were made.Pathological changes of colon were observed under light microscope and evaluated by histopathology of colon.The activity of MPO was detected by chemical colorimetry,and the expression of IL-6 and TGF-βin serum was detected by ELISA.Results:Compared with the control group,the general situation of the model group was worse,such as color,activity,occult blood degree,fecal traits;DAI was the largest;pathological damage was obvious;HS scores were the most serious and the expression of serum IL-6 and TGF-βand colon MPO was significantly increased.After intervention,the above indices in Qingchang Wenzhong Decoction group,the transplantation group and the combination group were improved,and the expression of serum IL-6 and TGF-βand colon MPO were down-regulated,with statistical significance(P<0.05 or P<0.01).In improving the occult blood in feces,the combination group was significantly better than Qingchang Wenzhong Decoction group and the transplantation group(P<0.05 or P<0.01).Conclusion:Qingchang Wenzhong Decoction,fecal bacteria transplantation and their combination can significantly alleviate the symptoms of UC rats induced by DSS,and may play a role by down-regulating the expression of IL-6 and TGF-β.The effect of combined application on improving hematochezia is better than single therapeutic method.