OBJECTIVE: This work uses an aqueous extract from C. sylvestris var. lingua leaves (AECs) to evaluate the acute anti-ulcerative colitis efficacy in rat and HT-29 (human colorectal cancer cell line) models.
METHODS: To determine the secondary metabolites of AECs, liquid chromatography with a diode array detector (LC-DAD) study was carried out. 2,4,6-trinitrobenzenesulfonic acid (TNBS, 30 mg/0.25 mL EtOH 30% v/v) was used as an enema to cause acute colitis. Three days were spent giving the C. sylvestris var. lingua extract orally by gavage at dosages of 3, 30, and 300 mg/kg. The same route was used to deliver distilled water to the vehicle and naïve groups. After the animals were sacrificed on the fourth day, intestinal tissues were taken for histological examination and evaluation of biochemical tests such as those measuring superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), malondialdehyde (MDA), nitrite/nitrate, myeloperoxidase (MPO) activity. Additionally, interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and interleukin 10 (IL-10), were conducted on the intestinal tissues. Additionally, an MTT assay was used to evaluate the effect of AECs on the viability of HT-29 cells. Additionally, a molecular docking study was carried out to compare some potential target proteins with identified chemicals found in AECs.
RESULTS: LC-DAD analysis identified five compounds (caffeic acid, ellagic acid, ferulic acid, gallic acid, and quercetin) in AECs. Pre-administration of AECs (3; 30; 300 mg/kg) and mesalazine (500 mg/kg) reduced macroscopic scores (55%, 47%, 45%, and 52%, p < 0.001) and ulcerated areas (70.3%, 70.5%, 57%, and 56%, p < 0.001), respectively. It also increased SOD, GSH, and CAT activities (p < 0.01), while decreasing MDA (p < 0.001), nitrite/nitrate (p < 0.05), and MPO (p < 0.001) activities compared to the colitis group. Concerning inflammatory markers, significant modulations were observed: AECs (3, 30, and 300 mg/kg) lowered levels of IL-1β and TNF-α (p < 0.001) and increased IL-10 levels (p < 0.001) compared to the colitis groups. The viability of HT-29 cells was suppressed by AECs with an IC50 of 195.90 ± 0.01 μg/mL (48 h). During the molecular docking analysis, quercetin, gallic acid, ferulic acid, caffeic acid, and ellagic acid demonstrated consistent binding affinities, forming stable interactions with the 3w3l (TLR8) and the 3ds6 (MAPK14) complexes.
CONCLUSIONS: These results imply that the intestinal mucogenic, anti-inflammatory, and antioxidant properties of the C. sylvestris var. lingua leaf extract may be involved in its therapeutic actions for ulcerative colitis. The results of the in silico study point to the possibility of quercetin and ellagic acid interacting with P38 and TLR8, respectively, in a beneficial way.
目的:这项工作使用了来自C.sylvestrisvar的水提取物。lingua叶(AECs)评估大鼠和HT-29(人结直肠癌细胞系)模型中的急性抗溃疡性结肠炎功效。
方法:为了确定AECs的次级代谢产物,进行了二极管阵列检测器(LC-DAD)的液相色谱研究。2,4,6-三硝基苯磺酸(TNBS,30mg/0.25mLEtOH30%v/v)用作灌肠剂,以引起急性结肠炎。花了三天的时间给卷心菜。通过管饲法以3、30和300mg/kg的剂量口服lingua提取物。使用相同的途径将蒸馏水递送至载体和初始组。在第四天处死动物后,肠组织进行组织学检查和生化测试的评估,如测量超氧化物歧化酶(SOD),还原型谷胱甘肽(GSH),过氧化氢酶(CAT),丙二醛(MDA),亚硝酸盐/硝酸盐,髓过氧化物酶(MPO)活性。此外,白细胞介素1β(IL-1β),肿瘤坏死因子α(TNF-α),和白细胞介素10(IL-10),在肠组织上进行。此外,采用MTT法评价AECs对HT-29细胞活力的影响。此外,我们进行了一项分子对接研究,以将一些潜在的靶蛋白与AECs中发现的已鉴定的化学物质进行比较.
结果:LC-DAD分析确定了五种化合物(咖啡酸,鞣花酸,阿魏酸,没食子酸,和槲皮素)在AECs中。预先给药AECs(3;30;300mg/kg)和美沙拉嗪(500mg/kg)降低了宏观评分(55%,47%,45%,52%,p<0.001)和溃疡面积(70.3%,70.5%,57%,56%,p<0.001),分别。它也增加了SOD,GSH,和CAT活性(p<0.01),在降低MDA(p<0.001)的同时,亚硝酸盐/硝酸盐(p<0.05),与结肠炎组相比,MPO(p<0.001)活性。关于炎症标志物,观察到显著的调节:与结肠炎组相比,AECs(3,30和300mg/kg)降低IL-1β和TNF-α水平(p<0.001),并增加IL-10水平(p<0.001).HT-29细胞的活力被AECs抑制,IC50为195.90±0.01μg/mL(48小时)。在分子对接分析中,槲皮素,没食子酸,阿魏酸,咖啡酸,和鞣花酸表现出一致的结合亲和力,与3w3l(TLR8)和3ds6(MAPK14)复合物形成稳定的相互作用。
结论:这些结果暗示肠粘膜,抗炎,和C.sylvestrisvar的抗氧化性能。舌叶提取物可能参与其对溃疡性结肠炎的治疗作用。计算机研究的结果表明槲皮素和鞣花酸分别与P38和TLR8相互作用的可能性,以一种有益的方式。