华蟾素对FOLFOX化疗小鼠肠道炎症及黏膜屏障的影响

Effect of Huachansu on intestinal inflammation and mucosal barrier in FOLFOX chemotherapy mice

目的观察华蟾素对FOLFOX化疗小鼠肠道炎症及黏膜屏障的影响,探讨华蟾素缓解化疗相关性腹泻(CID)的机制。方法将28只BALB/c小鼠随机分为4组:正常组,FOLFOX组,华蟾素低剂量组(260 mg·kg^(-1)),华蟾素高剂量组(520 mg·kg^(-1))。FOLFOX组予以FOLFOX化疗方案连续腹腔注射5 d,华蟾素低、高剂量组给予FOLFOX化疗及灌胃相应剂量的华蟾素,正常组则予以等体积的生理盐水。记录小鼠的体重、小肠长度和腹泻评分,以苏木精-伊红(HE)染色观察小肠黏膜病理改变,以酶联免疫吸附法检测血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、D-乳酸(D-LA)、二胺氧化酶(DAO)、内毒素(ET)水平,以实时荧光定量聚合酶链反应法检测小肠黏膜中Occludin、Claudin-1 mRNA表达水平,以蛋白质印迹法检测小肠黏膜中NOD样受体蛋白3(NLRP3)、p-核转录因子-κB(p-NFκB)蛋白的表达水平。结果正常组、FOLFOX组和华蟾素低、高剂量组血清中D-LA的水平分别为(5.25±1.21),(10.29±1.15),(8.04±0.75)和(8.42±1.08)μmol·L^(-1);DAO水平分别为(244.36±42.50),(390.88±47.11),(293.78±37.57)和(318.22±54.26)pg·mL^(-1);ET水平分别为(8.34±2.12),(15.67±2.28),(11.46±1.83)和(12.33±2.12)EU·mL^(-1);IL-6水平分别为(16.41±2.98),(28.27±4.24),(16.32±2.45)和(22.08±3.77)pg·mL^(-1);IL-1β水平分别为(14.57±2.75),(24.20±3.20),(18.81±1.86)和(18.75±2.60)pg·mL^(-1);TNF-α水平分别为(83.54±14.51),(129.82±8.30),(98.59±17.55)和(110.11±9.81)pg·mL^(-1)。上述指标,FOLFOX组与正常组比较,华蟾素低剂量组与FOLFOX组比较,差异均有统计学意义(均P<0.05);华蟾素高剂量组与FOLFOX组比较,除NLRP3蛋白表达的相对水平外,其余指标差异均有统计学意义(P<0.05)。结论华蟾素能够缓解CID,抑制小肠黏膜炎症,保护肠黏膜屏障,其机制可能与抑制NLRP3/NF-κB信号通路有关。

Objective To observe the effect of Huachansu on the intestinal inflammation and mucosal barrier of FOLFOX chemotherapy mice,and to explore its mechanism in relieving chemotherapy induced diarrhea (CID).Methods Twenty-eight BALB/c mice were randomly divided into 4 groups:normal group,FOLFOX group,low-dose Huachansu group (260 mg·kg^(-1)),high-dose Huachansu group (520mg·kg^(-1)).FOLFOX group was given FOLFOX chemotherapy by intraperitoneal injection for 5 days,the low-dose Huachansu and high-dose Huachansu group were given the FOLFOX chemotherapy and corresponding dose of Huachansu by gavage,normal group was given equal volume of normal saline.The weight,length of the small intestine and diarrhea score of the mice were recorded.Hematoxylineosin (HE) staining was used to observe the pathological changes of the small intestine mucosa.Enzyme-linked immunosorbent assay was used to detect the serum interleukin-6 (IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),D-lactic acid (D-LA),diamine oxidase (DAO),endotoxin (ET).Real-time quantitative polymerase chain reaction was used to detect the expression levels of Occludin and Claudin-1 mRNA in the small intestinal mucosa,and Western blot was used to detect the expression levels of NOD-like receptor protein 3(NLRP3) and nuclear transcription factor-κB(p-NFκB) protein in the small intestinal mucosa.Results The levels of serum D-LA in normal group,FOLFOX group,low-dose and high-dose Huachansu groups were(5.25±1.21),(10.29±1.15),(8.04±0.75) and (8.42±1.08)μmol·L^(-1),the levels of DAO were(244.36±42.50),(390.88±47.11),(293.78±37.57) and (318.22±54.26) pg·m L^(-1),the levels of ET were(8.34±2.12),(15.67±2.28),(11.46±1.83) and (12.33±2.12) EU·m L^(-1),the levels of IL-6 were(16.41±2.98),(28.27±4.24),(16.32±2.45) and (22.08±3.77) pg·m L^(-1);the levels of IL-1βwere(14.57±2.75),(24.20±3.20),(18.81±1.86) and (18.75±2.60) pg·m L^(-1),the levels of TNF-αwere(83.54±14.51),(129.82±8.30),(98.59±17.55) and (110.11±9.81) pg·m L^(-1).For the above indicators,the differences between FOLFOX group and normal group,between low-dose Huachansu group and FOLFOX group were statistically significant (P<0.05).Compared with FOLFOX group,the high-dose Huachansu group showed statistically significant differences in other indicators except for the relative level of NLRP3 protein expression(P<0.05).Conclusion Huachansu can relieve CID,inhibit mucosal inflammation of small intestinal,and protect the intestinal mucosal barrier,which may be related to the inhibition of the NLRP3/NF-κB signaling pathway.