Abstract:
Berberis vulgaris L. (Berberidaceae) is a shrub that has been widely used in European folk medicine as an anti-inflammatory and antimicrobial agent. The purpose of our study was to elucidate the mechanisms of the chemopreventive action of the plant\'s methanolic root extract (BVR) against colon cancer cells. Studies were conducted in human colon adenocarcinoma cell lines (LS180 and HT-29) and control colon epithelial CCD841 CoN cells. According to the MTT assay, after 48 h of cell exposure, the IC50 values were as follows: 4.3, 46.1, and 50.2 µg/mL for the LS180, HT-29, and CCD841 CoN cells, respectively, showing the greater sensitivity of the cancer cells to BVR. The Cell Death Detection ELISAPLUS kit demonstrated that BVR induced programmed cell death only against HT-29 cells. Nuclear double staining revealed the great proapoptotic BVR properties in HT-29 cells and subtle effect in LS180 cells. RT-qPCR with the relative quantification method showed significant changes in the expression of genes related to apoptosis in both the LS180 and HT-29 cells. The genes BCL2L1 (126.86-421.43%), BCL2L2 (240-286.02%), CASP3 (177.19-247.83%), and CASP9 (157.99-243.75%) had a significantly elevated expression, while BCL2 (25-52.03%) had a reduced expression compared to the untreated control. Furthermore, in a panel of antioxidant tests, BVR showed positive effects (63.93 ± 0.01, 122.92 ± 0.01, and 220.29 ± 0.02 mg Trolox equivalents (TE)/g in the DPPH•, ABTS•+, and ORAC assays, respectively). In the lipoxygenase (LOX) inhibition test, BVR revealed 62.60 ± 0.87% of enzyme inhibition. The chemical composition of BVR was determined using a UHPLC-UV-CAD-MS/MS analysis and confirmed the presence of several known alkaloids, including berberine, as well as other alkaloids and two derivatives of hydroxycinnamic acid (ferulic and sinapic acid hexosides). The results are very promising and encourage the use of BVR as a comprehensive chemopreventive agent (anti-inflammatory, antioxidant, and pro-apoptotic) in colorectal cancer, and were widely discussed alongside data from the literature.
摘要:
小檗属(小檗科)是一种灌木,已在欧洲民间医学中广泛用作抗炎和抗菌剂。我们研究的目的是阐明植物的甲醇根提取物(BVR)对结肠癌细胞的化学预防作用的机制。在人结肠腺癌细胞系(LS180和HT-29)和对照结肠上皮CCD841CoN细胞中进行研究。根据MTT测定,细胞暴露48小时后,LS180、HT-29和CCD841CoN细胞的IC50值如下:4.3、46.1和50.2µg/mL,分别,显示癌细胞对BVR更敏感。细胞死亡检测ELISAPLUS试剂盒证明BVR仅针对HT-29细胞诱导程序性细胞死亡。核双染色显示HT-29细胞中的促凋亡BVR特性和LS180细胞中的微妙作用。使用相对定量方法的RT-qPCR显示LS180和HT-29细胞中与凋亡相关的基因表达的显着变化。基因BCL2L1(126.86-421.43%),BCL2L2(240-286.02%),CASP3(177.19-247.83%),CASP9(157.99-243.75%)表达显著升高,而与未处理的对照相比,BCL2(25-52.03%)具有降低的表达。此外,在一组抗氧化剂测试中,BVR显示出积极作用(DPPH•中的63.93±0.01、122.92±0.01和220.29±0.02mgTrolox当量(TE)/g,ABTS•+,和ORAC检测,分别)。在脂氧合酶(LOX)抑制试验中,BVR显示62.60±0.87%的酶抑制作用。使用UHPLC-UV-CAD-MS/MS分析确定BVR的化学组成,并确认存在几种已知的生物碱,包括小檗碱,以及其他生物碱和羟基肉桂酸的两种衍生物(阿魏酸和芥子酸己糖苷)。结果非常有希望,并鼓励使用BVR作为综合化学预防剂(抗炎,抗氧化剂,和促凋亡)在结直肠癌中,并与文献中的数据一起进行了广泛的讨论。