健脾益肾泄浊汤大鼠灌肠的含药血清对脂多糖诱导人结肠上皮细胞炎症损伤模型的影响

Effect of drug-containing serum of Jianpi Yishen Xiezhuo decoction on LPS-induced inflammatory injury of human colonic epithelial cells

目的通过观察健脾益肾泄浊汤大鼠灌肠的含药血清对人结肠上皮细胞炎症模型的干预效果,探讨该方灌肠对肠道机械屏障的影响。方法40只SPF级SD大鼠随机分为空白组、低剂量中药灌肠组、中剂量中药灌肠组、高剂量中药灌肠组,每组10只。空白组不予灌肠,低、中、高剂量组按1∶2∶4分别予0.4、0.8、1.6 mL中药灌肠,每次保留30 min,每天早晚2次,疗程5 d,于末次灌肠1 h后腹主动脉取血,离心后收集上层血清。人正常结肠上皮细胞(NCM460)分为空白组、模型组、低剂量中药灌肠血清组、中剂量中药灌肠血清组、高剂量中药灌肠血清组;除空白组外,其余各组均加入脂多糖(LPS)100μg/mL诱导细胞炎症损伤,中药灌肠血清组将细胞培养液更换为10%含药血清,模型组加入空白组大鼠血清,各组孵育24 h后检测细胞上清Claudin-1、Occludin、ZO-1、OD值、IL-6、TNF-α。结果与空白血清干预相比,中药灌肠含药血清可增加炎症损伤细胞的OD值,但不同剂量之间的作用效果未见差异(P>0.05);与空白血清干预相比,中药灌肠含药血清可降低炎症损伤细胞的上清IL-6及TNF-α水平,且高剂量的作用效果更为显著(P<0.05);与空白血清干预相比,中药灌肠的含药血清可增加炎症损伤细胞的Claudin-1、Occludin及ZO-1相对表达量,且高剂量的作用效果更为显著(P<0.05)。结论健脾益肾泄浊汤大鼠灌肠的含药血清可以增强肠上皮细胞活性,增加肠上皮细胞间紧密连接相关蛋白的表达,可能有助于保护肠道机械屏障功能,从而减轻局部炎症状态。

Objective To observe the intervention effect of drug-containing serum of Jianpi Yishen Xiezhuo decoction on human colonic epithelial cell inflammation model,and explore the effect of Jianpi Yishen Xiezhuo decoction on intestinal mechanical barrier.Methods 40 SPF SD rats were randomly divided into blank group(n=10),low dose TCM herb enema group(n=10),middle dose traditional Chinese medicine enema group(n=10)and high dose traditional Chinese medicine enema group(n=10).The blank group was not given enema,and the low,middle and high dose groups were given 0.4mL,0.8mL and 1.6mL traditional Chinese medicine enema at 1∶2∶4 for 30 minutes each time,twice in the morning and evening every day for 5 days.One hour after the last enema,blood was taken from the abdominal aorta,and the upper serum was collected after centrifugation.Human normal colonic epithelial cells(NCM460)were divided into blank group,model group,low-dose TCM herb enema serum group,medium-dose TCM herb enema serum group and high-dose TCM herb enema serum group.Except for the blank group,the other groups were added with 100μg/mL to induce cell inflammatory injury.The TCM herb enema serum group replaced the cell culture medium with 10%drug-containing serum,and the model group was added with the rat serum of the blank group.After incubation for 24 hours,the supernatant of cells were detected for Claudin-1,Occludin,ZO-1,OD,IL-6,TNF-α.Results Compared with blank serum intervention,TCM herb enema drug-containing serum could enhance the OD value of inflammatory injured cells,but there was no difference among different doses,compared with blank serum intervention,TCM herb enema drug-containing serum could reduce the levels of IL-6 and TNF-αin the supernatant of inflammatory injured cells,and the effect of high dose was more significant.Compared with the blank serum intervention,the drug-containing serum of TCM herb enema can increase the relative expression of Claudin-1,Occludin and ZO-1 of inflammatory injury cells,and the effect of high dose is more significant.Conclusion The drug-containing serum of Jianpi Yishen Xiezhuo decoction can enhance the activity of intestinal epithelial cells and increase the expression of tight junction-related proteins between intestinal epithelial cells,which may help to protect the intestinal mechanical barrier function and reduce local inflammation.