温肾健脾法对结肠癌癌前病变大鼠肠道菌群结构的影响及作用机制研究

Warming Kidney and Strengthening Spleen for Intestinal Flora Structure of Rats with Precancerous Lesions of Colon Cancer and Its Mechanism

目的探讨温肾健脾法在结肠癌癌前病变大鼠中的作用机制及对肠道菌群结构的影响。方法购置Wistar大鼠75只,随机取Wistar大鼠15只设为对照组;剩余60只Wistar大鼠腹腔注射DMH制备大鼠结肠癌癌前病变(异常肠腺病灶,ACF)模型,建模成功后随机数字表法分为模型组、高剂量组、中剂量组和低剂量组,每组15只。模型组和对照组予等体积生理盐水灌胃,高剂量组(3.10 g/kg灌胃)、中剂量组(1.55 g/kg灌胃)和低剂量组(0.775 g/kg灌胃)给予温肾健脾法干预,比较各组肠道菌群结构、HE染色及分泌型免疫球蛋白(sIgA)、血清D乳酸、白细胞介素-17(IL-17)水平。结果中剂量组造模第2、6、17周ACF水平均低于低剂量组和高剂量组(P<0.05);HE染色结果表明:低剂量组黏膜轻到中度慢性炎,局灶淋巴细胞聚集伴挤压,淋巴组织增生,局灶腺体轻度异型增生,个别黏膜腺癌形成。中剂量组黏膜轻度慢性炎,淋巴滤泡形成,未见异型增生及癌形成。高剂量组:黏膜轻-中度慢性炎,伴有轻到中度异型增生;中剂量组sIgA水平高于低剂量组与高剂量组(P<0.05);血清D乳酸、IL-17水平低于低剂量组与高剂量组(P<0.05)。结论中剂量温肾健脾法用于结肠癌癌前病变大鼠中能改善肠道菌群结构,可调节sIgA、血清D乳酸、IL-17水平,减轻肠黏膜结构损伤及异型增生。

Objective To explore the mechanism of warming kidney and strengthening spleen method in colon cancer precancerous lesions and its effect on intestinal flora structure.Methods Seventy-five Wistar rats were purchased,and 15 Wistar rats were randomly selected as a control group.The remaining 60 Wistar rats were intraperitoneally injected with DMH to prepare a model of colon cancer precancerous lesions(abnormal intestinal gland lesions,ACF)model.After successful modeling,according to the random number table method,the rats were divided into model group,high-dose group,middle-dose group and low-dose group,15 animals in each group.The model group and the control group were gavaged with equal volume of normal saline.The high-dose group(3.10 g/kg,gavage),the middle-dose group(1.55 g/kg,gavage)and the low-dose group(0.775 g/kg,gavage)were given the formula of warming kidney and strengthening spleen method.The intestinal flora structure,HE staining and secreted immunoglobulin(sIgA),serum D lactate and interleukin-17(IL-17)levels in each group were compared.Results In the middle-dose group,the ACF level was lower than that in the low-dose group and the high-dose group at week 2,6,and 17(P<0.05).HE staining showed that the low-dose group had mild to moderate chronic inflammation and focal lymphocyte aggregation with squeezing,lymphoid tissue hyperplasia,focal gland dysplasia,and formation of individual mucosal adenocarcinoma.Mild chronic inflammation of the mucosa,lymphoid follicle formation,no dysplasia or cancer formation were in the middle-dose group.High-dose group had mild-to-moderate chronic inflammation of the mucosa with mild to moderate dysplasia.The sIgA levels in the middle-dose group were higher than those in the low-dose group and the high-dose group(P<0.05).The serum D lactate and IL-17 levels in the middle-dose group were lower than those in the low-dose group and the high-dose group(P<0.05).Conclusion The middle-dose group of warming kidney and strengthening spleen method can improve the structure of intestinal flora,regulate the levels of sIgA,serum D lactate and IL-17,and alleviate structural damage and dysplasia of intestinal mucosa in the treatment of colon cancer precancerous lesions.