人工蝉花对5-氟尿嘧啶诱发大鼠肠黏膜损伤的修复作用研究

Study on Repair Effect of Artificial Isaria cicadae on Intestinal Mucosal Injury Induced by 5-Fluorouracil in Rats

目的:研究人工蝉花对5-氟尿嘧啶(5-FU)诱发大鼠肠黏膜损伤的修复作用。方法:将40只SD大鼠随机分为正常组(生理盐水)、模型组(生理盐水)和人工蝉花高、中、低剂量(3.5、1.75、0.875 g/kg)组,每组8只。除正常组外,其余各组大鼠每日腹腔注射30 mg/kg的5-FU(0.25 g/10 m L)1次,连续5 d;同时,各组大鼠每天灌胃相应药物/生理盐水1次,连续给药8 d。给药结束后,测定各组大鼠体质量变化,苏木精-伊红染色后观察各组大鼠小肠组织病理学变化,并检测各组大鼠生物屏障相关因子[血清中内毒素(ET)、D-乳酸(D-LA)]水平和免疫屏障相关因子[血清中肿瘤坏死因子α(TNF-α)、γ-干扰素(IFN-γ)、分泌性免疫球蛋白(sIgA)、白细胞介素15(IL-15)、集落刺激因子(G-CSF)和小肠组织中髓过氧化物酶(MPO)、丙二醛(MDA)]水平以及机械屏障相关因子(肠表面闭锁小带蛋白ZO-1和紧密连接蛋白Claudin-1)水平。结果:与正常组比较,模型组大鼠体质量显著减少(P<0.01);肠绒毛剥落明显,隐窝结构散乱,大量炎性细胞聚集,肠黏膜损伤严重;血清中D-LA、ET和TNF-α、IFN-γ、MPO、G-CSF水平以及小肠组织中MDA水平均显著升高(P<0.01);血清中sIgA、IL-15水平以及小肠组织中ZO-1、Claudin-1蛋白表达水平显著降低(P<0.05或P<0.01)。与模型组比较,人工蝉花高剂量组大鼠体质量均显著增加(P<0.01);各给药组大鼠小肠的病理变化均得到不同程度改善,其中人工蝉花高、中剂量组小肠形态接近正常组;各给药组大鼠血清中D-LA、ET和TNF-α、IFN-γ、MPO、G-CSF水平以及小肠组织中MDA水平均显著降低(P<0.01);各给药组大鼠血清中sIgA、IL-15水平以及人工蝉花高、中剂量组大鼠小肠组织中Claudin-1、ZO-1蛋白表达水平均显著升高(P<0.01)。结论:人工蝉花可从机械屏障、免疫屏障、生物屏障等多个方面修复5-FU所致大鼠肠黏膜损伤。

OBJECTIVE:To study the repair effect of artificial Isaria cicadae on intestinal mucosal injury induced by5-fluorouracil(5-FU)in rats.METHODS:Forty SD rats were randomly divided into normal group(normal saline),model group(normal saline),artificial I.cicadae high-dose,medium-dose and low-dose groups(3.5,1.75,0.875 g/kg),with 8 rats in each group.Except for normal group,other groups were given intraperitoneal injection of 5-FU(0.25 g/10 mL)30 mg/kg,once a day,for consecutive 5 days.At the same time,each group was given relevant medicine/normal saline intragastrically,once a day,for consecutive 8 d.After medication,body weight of rat was determined in each group.HE staining was used to observe the pathological change of small intestine.The levels of biobarrier-related factor[endotoxin(ET),D-lactic acid(D-LA)],immune barrier related factors(TNF-α,IFN-γ,sIgA,IL-15,G-CSF in serum and MPO,MDA in small intestine)and the levels of mechanical barrier related factors(connexin ZO-1 and Claudin-1)were detected.RESULTS:Compared with normal group,body weight of rats in model group was decreased significantly(P<0.01).Intestinal villus exfoliated obviously,the crypt structure was scattered,a large number of inflammatory cells gathered,and intestinal mucosa was seriously damaged.Serum levels of ET and D-LA,the levels of TNF-a,IFN-γ,MPO and G-CSF in serum,MDA level in small intestine were increased significantly(P<0.01).Serum levels of sIgA and IL-15 as well as the expressions of ZO-1 and Claudin-1 in small intestine were decreased significantly(P<0.05 or P<0.01).Compared with model group,body weight of rats in artificial I.cicadae high-dose group was increased significantly(P<0.01).The pathological changes of the small intestine of rats in each administration group were improved to varying degrees.The intestine morphology of artificial I.cicadae high-dose and medium-dose groups was close to that of the normal group.The levels of and ET,D-LA,TNF-α,IFN-γ,MPO,G-CSF in serum and the level of MDA in intestinal were decreased significantly(P<0.01).Serum levels of sIgA and IL-15 in administration groups as well as the expressions of ZO-1 and Claudin-1 in intestinal tissue were increased significantly(P<0.01).CONCLUSIONS:Artificial I.cicadae can repair intestinal mucosal damage caused by 5-FU in respects of mechanical barrier,immune barrier,biological barrier.