Gentianopsis paludosa xanthone combined with probiotics inhibits colon inflammation-tumor transformation in rats by regulating TGF-β1/Smads pathway and inflammatory factors

doi:10.11958/20230462

Abstract

Abstract:

Objective To investigate the mechanism of Gentianopsis paludosa xanthone (GPX) combined with probiotics in the intervention of colon inflammation-tumor transformation in rats by regulating TGF-β1/Smads pathway and inflammatory factors. Methods Ninety rats were divided into the normal group, the model group [drinking sodium dextran sulfate (DSS) for 3 days] and the intervention group by random number table method. The model group was subdivided into the inflammatory stage group, the pre-inflammatory cancer group (DMH injection for 4 weeks), the intermediate inflammatory cancer group (DMH injection for 13 weeks) and the advanced inflammatory cancer group (DMH injection for 21 weeks). The administration group was subdivided into the groups (after the first day of drinking DSS, drugs for each group were given by gavage once a day for 8 weeks) on the basis of the advanced inflammatory cancer group, including the GPX group (GPX 69.3 mg/kg), the probiotic group, the combined group (GPX+probiotics 400 mg/kg) and the thalidomide group (thalidomide 13.5 mg/kg). The disease activity index (DAI), colon length and wet mass index were compared between all groups. Characteristics of colon tumors were observed, and pathological changes of colon were observed by HE staining. The expression levels of transforming growth factor (TGF) -β1, Smad4, Smad7, interleukin (IL)-6 and tumor necrosis factor (TNF) -α were detected by Western blot assay and enzyme-linked immunosorbent assay, respectively. Results Compared with the advanced inflammatory cancer group, the administration groups showed an increase in colon length, the expression levels of TGF-β1 and Smad4 protein, a decrease in colon wall thickness, wet mass index, maximum tumor diameter, the levels of Smad7, IL-6, TNF-α, and DAI score decreased in the GPX group and the combined group (P<0.05). The structure and morphology of intestinal mucosa were improved in the GPX group, the probiotic group and the combination group, and the structure of colonic crypt and goblet cell number were increased. Compared with the probiotic group and the GPX group, the colon wall thickness, colon wet mass index and tumor number were decreased, the protein expression levels of TGF-β1 and Smad4 were increased, and levels of IL-6 and TNF-α were decreased in the combination group (P<0.05). Conclusion GPX combined with probiotics could inhibit the transformation of colon inflammation-tumor, and the mechanism may be related to the regulation of TGF-β1/Smads pathway and the inhibition of pro-inflammatory factors of IL-6 and TNF-α.

Key words: colorectal neoplasms, colitis, biotransformation, Gentianopsis paludosa xanthone, probiotics, TGF-β1/Smads pathway

CLC Number:

R735.35

Figures/Tables 7

References 16

[1]	张聪伟, 刘金响, 吕小龙, 等. 白头翁汤灌肠辅助治疗溃疡性结肠炎的有效性及安全性的Meta分析[J]. 中国中西医结合外科杂志, 2023, 29(6):758-765.
	ZHANG C W, LIU J X, LV X L, et al. Baitouweng decoction enema for ulcerative colitis:Meta-analysis[J]. Chinese Journal of Surgery of Integrated Traditional and Western Medicine, 2023, 29(6):758-765. doi:10.3969/j.issn.1007-6948.2023.05.007.
[2]	ZHANG W, FU X, XIE J, et al. miR-26a attenuates colitis and colitis-associated cancer by targeting the multiple intestinal inflammatory pathways[J]. Mol Ther Nucleic Acids, 2021, 24:264-273. doi:10.1016/j.omtn.2021.02.029.
[3]	CHAURIO R A, ANADON C M, LEE COSTICH T, et al. TGF-β-mediated silencing of genomic organizer SATB1 promotes Tfh cell differentiation and formation of intra-tumoral tertiary lymphoid structures[J]. Immunity, 2022, 55(1):115-128.e9. doi:10.1016/j.immuni.2021.12.007.
[4]	MEANS A L, FREEMAN T J, ZHU J, et al. Epithelial Smad4 deletion up-regulates inflammation and promotes inflammation-associated cancer[J]. Cell Mol Gastroenterol Hepatol, 2018, 6(3):257-276. doi:10.1016/j.jcmgh.2018.05.006.
[5]	KHATTAB A E, DARWISH A M, OTHMAN S I, et al. Anti-inflammatory and immunomodulatory potency of selenium-enriched probiotic mutants in mice with induced ulcerative colitis[J]. Biol Trace Elem Res, 2023, 201(1):353-367. doi:10.1007/s12011-022-03154-1.
[6]	LU N H, ZHAO H Q, JING M, et al. The pharmacodynamic active components study of Tibetan medicine Gentianopsis paludosa on ulcerative colitis fibrosis[J]. Int Immunopharmacol, 2017, 46:163-169. doi:10.1016/j.intimp.2017.01.001.
[7]	高娜, 景明, 张艳霞, 等. 藏药湿生扁蕾对人结肠癌SW480细胞凋亡调控因子Bcl-2及Bax的mRNA表达的影响[J]. 中国民族医药杂志, 2014, 20(9):51-53.
	GAO N, JING M, ZHANG Y X, et al. The effect of Tibetan Medicine Gentianopsis Paludosa about colon cancer SW480 cell apoptosis regulatory factors mRNA expression of Bcl-2 and Bax[J]. Journal of Medicine & Pharmacy of Chinese Minorities, 2014, 20(9):51-53. doi:10.16041/j.cnki.cn15-1175.2014.09.033.
[8]	寇亮, 刘越敏, 柳娜, 等. 大孔吸附树脂分离纯化湿生扁蕾总𠮿酮[J]. 中药新药与临床药理, 2022, 33(4):522-530.
	KOU L, LIU Y M, LIU N, et al. Separation and purification of total xanthones from Gentianopsis paludosa using macroporous adsorption resin[J]. Tradit Chin Drug Res Clin Pharmacol, 2022, 33(4):522-530. doi:10.19378/j.issn.1003-9783.2022.04.014.
[9]	KLITGAARD M, KRISTENSEN M N, VENKATASUBRAMANIAN R, et al. Assessing acute colitis induced by dextran sulfate sodium in rats and its impact on gastrointestinal fluids[J]. Drug Deliv Transl Res, 2023, 13(5):1484-1499. doi:10.1007/s13346-023-01313-y.
[10]	卢年华, 芦彦兆, 赵慧巧, 等. 藏药湿生扁蕾与獐牙菜的相关本草考证[J]. 中国药房, 2016, 27(19):2647-2649.
	LU N H, LU Y Z, ZHAO H Q, et al. Related herbal textual research on Tibetan herb Shi Sheng Bian Lei and Zhang Ya Cai[J]. China Pharm, 2016, 27(19):2647-2649. doi:10.6039/j.issn.1001-0408.2016.19.18.
[11]	寇亮. 湿生扁蕾总𠮿酮抗UC肠纤维化机制研究及其结肠定位微丸的制备[D]. 兰州: 甘肃中医药大学, 2022.
	KOU L. Study on the mechanism of anti-UC intestinal fibrosis by total xanthones from Gentianopsis paludosa and preparation of colon-specific[D]. Lanzhou: Gansu University of Traditional Chinese Medicine, 2022. doi:10.27026/d.cnki.ggszc.2022.000003.
[12]	柳娜, 刘越敏, 薄双琴, 等. 湿生扁蕾总𠮿酮对NCM460细胞间质转化的影响[J]. 中国细胞生物学学报, 2022, 44(9):1735-1743.
	LIU N, LIU Y M, BO S Q, et al. Effect of xanthones from Gentianopsis paludosa on mesenchymal transformation of NCM460 cells[J]. Chinese Journal of Cell Biology, 2022, 44(9):1735-1743. doi:10.11844/cjcb.2022.09.0005.
[13]	TRONCONE E, MARAFINI I, STOLFI C, et al. Involvement of Smad7 in inflammatory diseases of the gut and colon cancer[J]. Int J Mol Sci, 2021, 22(8):3922. doi:10.3390/ijms22083922.
[14]	TAGLIARI E, CAMPOS L F, CASAGRANDE T, et al. Effects of oral probiotics administration on the expression of transforming growth factor β and the proinflammatory cytokines interleukin 6, interleukin 17, and tumor necrosis factor α in skin wounds in rats[J]. JPEN J Parenter Enteral Nutr, 2022, 46(3):721-729. doi:10.1002/jpen.2216.
[15]	JÓSA V, FÉDERER K, ZRUBKA Z, et al. Investigation of IL6 expression in patients with colorectal adenocarcinoma[J]. Orv Hetil, 2021, 162(37):1502-1507. doi:10.1556/650.2021.32206.
[16]	BAKSHI H A, QUINN G A, NASEF M M, et al. Crocin inhibits angiogenesis and metastasis in colon cancer via TNF-α/NF-kB/VEGF pathways[J]. Cells, 2022, 11(9):1502. doi:10.3390/cells11091502.

组别	结肠长度/cm	壁厚/mm	结肠湿质量指数/（mg/mm）	DAI评分/分
正常组	16.13±0.71	0.529±0.019	13.11±0.78	0
炎症期组	15.68±0.54	0.518±0.007	24.82±0.91^a	2.67±0.47^a
炎癌前期组	15.10±0.99^a	0.545±0.009^a	19.42±0.57^a	0.47±0.17^a
炎癌中期组	13.20±0.88^a	1.648±0.087^a	34.54±1.19^a	1.50±0.36^a
炎癌末期组	11.53±0.58^a	2.099±0.104^a	40.73±1.09^a	2.07±0.47^a
益生菌组	12.88±1.66^b	1.117±0.009^b	32.42±1.35^b	2.03±0.19^a
GPX组	14.52±0.74^bc	1.001±0.021^bc	30.47±1.35^bc	1.67±0.16^bc
联合组	14.78±0.66^bc	0.824±0.007^bcd	28.93±1.46^bcd	1.53±0.32^bc
沙利度胺组	12.36±0.94^bd	1.418±0.011^bcd	33.46±1.68^bd	1.90±0.32
F	30.253^＊＊	1 413.920^＊＊	487.379^＊＊	71.192^＊＊

组别	结肠长度/cm	壁厚/mm	结肠湿质量指数/（mg/mm）	DAI评分/分
正常组	16.13±0.71	0.529±0.019	13.11±0.78	0
炎症期组	15.68±0.54	0.518±0.007	24.82±0.91^a	2.67±0.47^a
炎癌前期组	15.10±0.99^a	0.545±0.009^a	19.42±0.57^a	0.47±0.17^a
炎癌中期组	13.20±0.88^a	1.648±0.087^a	34.54±1.19^a	1.50±0.36^a
炎癌末期组	11.53±0.58^a	2.099±0.104^a	40.73±1.09^a	2.07±0.47^a
益生菌组	12.88±1.66^b	1.117±0.009^b	32.42±1.35^b	2.03±0.19^a
GPX组	14.52±0.74^bc	1.001±0.021^bc	30.47±1.35^bc	1.67±0.16^bc
联合组	14.78±0.66^bc	0.824±0.007^bcd	28.93±1.46^bcd	1.53±0.32^bc
沙利度胺组	12.36±0.94^bd	1.418±0.011^bcd	33.46±1.68^bd	1.90±0.32
F	30.253^＊＊	1 413.920^＊＊	487.379^＊＊	71.192^＊＊

组别	结肠瘤体最大直径/mm	结肠瘤体数量/个
正常组	0	0
炎症期组	0	0
炎癌前期组	0	0
炎癌中期组	9.88±0.23^a	7.80±0.79^a
炎癌末期组	13.14±0.69^a	9.90±0.57^a
益生菌组	4.91±0.26^b	4.70±0.67^b
GPX组	2.11±0.10^bc	3.50±0.53^bc
联合组	1.90±0.13^bc	1.80±0.63^bcd
沙利度胺组	5.08±0.14^bd	7.30±0.95^bcd
F	3 054.074^＊＊	429.213^＊＊

组别	结肠瘤体最大直径/mm	结肠瘤体数量/个
正常组	0	0
炎症期组	0	0
炎癌前期组	0	0
炎癌中期组	9.88±0.23^a	7.80±0.79^a
炎癌末期组	13.14±0.69^a	9.90±0.57^a
益生菌组	4.91±0.26^b	4.70±0.67^b
GPX组	2.11±0.10^bc	3.50±0.53^bc
联合组	1.90±0.13^bc	1.80±0.63^bcd
沙利度胺组	5.08±0.14^bd	7.30±0.95^bcd
F	3 054.074^＊＊	429.213^＊＊

组别	TGF-β1	Smad4	Smad7
正常组	0.87±0.05	0.93±0.02	0.47±0.06
炎症期组	0.45±0.04^a	0.56±0.03^a	0.57±0.09^a
炎癌前期组	0.58±0.05^a	0.70±0.02^a	0.68±0.08^a
炎癌中期组	0.34±0.05^ab	0.50±0.02^ab	0.88±0.10^ab
炎癌末期组	0.25±0.04^abc	0.35±0.03^abc	1.05±0.07^abc
益生菌组	0.76±0.07^d	0.70±0.03^d	0.83±0.01^d
GPX组	0.79±0.06^d	0.94±0.03^de	0.60±0.01^de
联合组	0.99±0.05^def	1.07±0.12^def	0.58±0.03^de
沙利度胺组	0.69±0.05^def	0.65±0.02^def	0.77±0.02^def
F	228.750^＊＊	259.751^＊＊	84.431^＊＊