湿生扁蕾呫吨酮联合益生菌调控TGF-β1/Smads通路及炎性因子干预大鼠结肠炎癌转化

doi:10.11958/20230462

天津医药 ›› 2024, Vol. 52 ›› Issue (2): 136-141.doi: 10.11958/20230462

湿生扁蕾呫吨酮联合益生菌调控TGF-β1/Smads通路及炎性因子干预大鼠结肠炎癌转化

卢年华(), 靳展洪叶, 张茜, 张萌, 李俊珂, 赵慧巧^△(), 张永鹏

河北北方学院中医学院（邮编075000）

收稿日期:2023-03-31 修回日期:2023-06-28 出版日期:2024-02-15 发布日期:2024-01-26
通讯作者: ^△ E-mail：1325478922@qq.com
作者简介:卢年华（1989），男，讲师，主要从事结肠炎癌转化研究。E-mail：18215174868@163.com
基金资助:
河北省自然科学基金资助项目(H2020405027);河北省自然科学基金资助项目(H2020405010);河北省高等学校科学技术研究项目(QN2020118);河北省属高校基本科研业务项目(JYT2021036);河北省属高校基本科研业务项目(JYT2019009)

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

LU Nianhua(), JIN Zhanhongye, ZHANG Qian, ZHANG Meng, LI Junke, ZHAO Huiqiao^△(), ZHANG Yongpeng

College of Traditional Chinese Medicine, Hebei North University, Zhangjiakou 075000, China

Received:2023-03-31 Revised:2023-06-28 Published:2024-02-15 Online:2024-01-26
Contact: ^△ E-mail: 1325478922@qq.com

卢年华, 靳展洪叶, 张茜, 张萌, 李俊珂, 赵慧巧, 张永鹏. 湿生扁蕾呫吨酮联合益生菌调控TGF-β1/Smads通路及炎性因子干预大鼠结肠炎癌转化[J]. 天津医药, 2024, 52(2): 136-141.

LU Nianhua, JIN Zhanhongye, ZHANG Qian, ZHANG Meng, LI Junke, ZHAO Huiqiao, ZHANG Yongpeng. Gentianopsis paludosa xanthone combined with probiotics inhibits colon inflammation-tumor transformation in rats by regulating TGF-β1/Smads pathway and inflammatory factors[J]. Tianjin Medical Journal, 2024, 52(2): 136-141.

摘要/Abstract

摘要：

目的基于TGF-β1/Smads通路及炎性因子探讨湿生扁蕾呫吨酮（GPX）联合益生菌干预结肠炎癌转化的机制。方法将90只大鼠采用随机数字表法分为正常组、模型组[饮用右旋糖酐硫酸钠（DSS）3 d]和给药组。其中，模型组又分为炎症期组、炎癌前期组[注射二甲基肼（DMH）4周]、炎癌中期组（注射DMH 13周）、炎癌末期组（注射DMH 21周）；给药组是在炎癌末期组的基础上边建模边给药且根据所给药物再分组，即饮用DSS 3 d，在饮用DSS的第1天就开始灌胃各组药物，每日1次，持续8周，分别为GPX 69.3 mg/kg（GPX组）、益生菌400 mg/kg（益生菌组）、GPX 69.3 mg/kg+益生菌400 mg/kg（联合组）、沙利度胺13.5 mg/kg（沙利度胺组）。对各组疾病活动指数（DAI）、结肠长度和湿质量指数进行比较；观察大鼠结肠瘤体特征；HE染色观察结肠病理变化；Western blot和酶联免疫吸附试验分别检测转化生长因子（TGF）-β1，Smad4，Smad7和白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α表达。结果与炎癌末期组比较，各给药组结肠长度增加，结肠壁厚、湿质量指数与瘤体最大直径降低，TGF-β1和Smad4蛋白表达水平升高，Smad7、IL-6、TNF-α水平降低，GPX组与联合组DAI得分降低（P<0.05），GPX组、益生菌组和联合组的肠黏膜层结构和形态改善，结肠隐窝结构与杯状细胞数量增加；与益生菌组和GPX组比较，联合组结肠壁厚、结肠湿质量指数、瘤体数量降低，TGF-β1和Smad4蛋白表达水平升高，IL-6、TNF-α水平降低（P<0.05）。结论 GPX和益生菌联用可抑制结肠炎癌转化，其作用机制可能与调控TGF-β1/Smads通路和抑制IL-6、TNF-α表达有关。

关键词: 结直肠肿瘤, 结肠炎, 生物转化, 湿生扁蕾呫吨酮, 益生菌, TGF-β1/Smads通路

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

中图分类号:

R735.35

图/表 7

参考文献 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^＊＊

湿生扁蕾呫吨酮联合益生菌调控TGF-β1/Smads通路及炎性因子干预大鼠结肠炎癌转化

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

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 7

参考文献 16

相关文章 15

编辑推荐

Metrics

本文评价

组别	IL-6	TNF-α
正常组	86.52±4.32	25.62±1.76
炎症期组	178.94±6.82^a	49.95±3.11^a
炎癌前期组	139.46±7.35^ab	39.43±2.36^ab
炎癌中期组	195.31±9.8^ac	54.55±2.83^ac
炎癌末期组	201.67±10.87^ac	62.62±1.65^ac
益生菌组	165.67±7.29^d	45.05±2.42^d
GPX组	152.55±6.94^de	41.90±2.39^de
联合组	143.11±6.37^def	38.54±2.67^def
沙利度胺组	171.05±9.67^df	55.48±2.55^def
F	190.906^＊＊	204.694^＊＊

[1]	王新波, 罗冰清, 石玉宝, 张也, 席江伟. 结直肠癌组织LncRNA LINC00342和miR-203a-3p表达及与预后的关系[J]. 天津医药, 2024, 52(9): 971-976.
[2]	陈静, 魏运姣, 罗超, 黄利华, 陈橙, 段莎莎. 基于Nrf2/ARE抗氧化应激途径探究乌梅丸对溃疡性结肠炎小鼠的作用机制[J]. 天津医药, 2024, 52(3): 278-254.
[3]	张婕, 苗耐英, 李欢, 刘新建. 奥曲肽对新生儿坏死性小肠结肠炎患儿粪便性状及体液免疫的影响[J]. 天津医药, 2024, 52(12): 1331-1334.
[4]	孙岩岩, 张骊, 陈池义, 蒋文涛. 肝移植治疗不可切除性结直肠癌肝转移的研究进展[J]. 天津医药, 2024, 52(12): 1335-1340.
[5]	张萌, 崔瑞洁, 李彩芳, 高琦, 李斐. 盐酸小檗碱片联合复方谷氨酰胺对溃疡性结肠炎患者肠黏膜屏障功能的影响[J]. 天津医药, 2024, 52(1): 102-106.
[6]	史刚刚, 郑冰, 韩梅. 溃疡性结肠炎患者发生阑尾口周围炎的危险因素及预后分析[J]. 天津医药, 2023, 51(4): 404-408.
[7]	柴小兵, 张利, 褚菲菲, 吴慧丽. m6A识别蛋白HuR调控lncRNA TRG-AS1抑制结直肠癌生长的机制研究[J]. 天津医药, 2023, 51(2): 124-130.
[8]	肖秋萍, 赵畅, 刘端勇, 李姗姗, 施旻, 陈丽玲, 钟友宝. 补脾益肠丸重塑CD4⁺T细胞亚群稳态缓解溃疡性结肠炎的机制研究[J]. 天津医药, 2023, 51(12): 1332-1338.
[9]	杨超, 张志永. 2型糖尿病患者结直肠腺瘤发病的多因素风险预测模型构建[J]. 天津医药, 2023, 51(1): 95-99.
[10]	邹大军, 胡福德, 周启立, 徐晓青. miR-301a对小鼠坏死性小肠结肠炎的影响及作用机制[J]. 天津医药, 2022, 50(9): 938-942.
[11]	邹夏慧, 刘敦菊, 万德惠, 姜慧英. 南昌市西湖区结直肠癌筛查结果及进展期结直肠腺瘤列线图预测模型的构建#br#[J]. 天津医药, 2022, 50(5): 528-532.
[12]	杨天骄, 柯创宏, 曾词正, 林丽曼, 胡海玲, 莫笑欢 . 腹部超声对新生儿食物蛋白诱导的小肠结肠炎综合征与早期新生儿坏死小肠结肠炎的鉴别价值[J]. 天津医药, 2022, 50(2): 210-213.
[13]	葛军涛, 詹江华△. 胆道闭锁Kasai术后辅助治疗研究进展[J]. 天津医药, 2022, 50(1): 30-34.
[14]	李娟, 袁向飞 , 赵煜 . 小肠结肠炎综合征小鼠肠黏膜中NLRP3表达与炎症分子关系的研究[J]. 天津医药, 2021, 49(8): 829-832.
[15]	郑磊 , 路佳 . 单细胞测序技术及其在结直肠癌中的应用[J]. 天津医药, 2021, 49(8): 891-896.