中草药

党参含药血清通过调控miR-21/PTEN轴改善溃疡性结肠炎

PDF下载

中草药 | 药理与临床 2026,57(5): 1773-1786

收起

中草药 |药理与临床 2026 , 57 (5) : 1773 -1786

党参含药血清通过调控miR-21/PTEN轴改善溃疡性结肠炎

全屏

王春霞¹, 杨洁², 史奇¹, 李芳¹, 陈正君¹, 杨彦平¹, 冯永辉³, 杨扶德¹

作者信息

通讯作者:

杨扶德

作者简介:

王春霞: 王春霞,博士研究生,主要从事中药品质研究。E-mail:1463120326@qq.com

Codonopsis Radix containing serum improves ulcerative colitis by regulating miR-21/PTEN axis

WANG Chunxia, YANG Jie, SHI Qi, LI Fang, CHEN Zhengjun, YANG Yanping, FENG Yonghui, YANG Fude

Affiliations

doi: 10.7501/j.issn.0253-2670.2026.05.016

文章导航

摘要

收起

目的探讨党参含药血清（Codonopsis Radix containing serum，CRCS）对溃疡性结肠炎（ulcerative colitis，UC）体外模型炎症因子释放及上皮细胞过度凋亡的作用机制。方法借助生物信息学分析工具预测调控磷酸酶和张力蛋白同源物（phosphatase and tensin homolog，PTEN）上游miRNA，采用qRT-PCR和Western blotting检测5% 2,4,6-三硝基苯磺酸（2,4,6-trinitrobenzenesulfonic acid solution，TNBS）/乙醇复合溶液灌肠诱导的UC大鼠模型结肠组织中PTEN mRNA和蛋白表达水平，以及miR-21、miR-30a-3p、miR-198、miR-152-3p、miR-301a-3p、miR-320、miR-3691-5p表达变化。10 μg/mL脂多糖（lipopolysaccharide，LPS）诱导人正常结肠上皮NCM460细胞构建UC体外模型，给予1%、2%、4%、6%、8% CRCS干预24 h，利用CCK-8法检测细胞活力。分别转染miR-21模拟剂（miRNA-21 mimics、miR-21 mimics）与miR-21抑制剂（miRNA-21 inhibitor、miR-21 inhibitor）至模型细胞，给予6% CRCS完全培养基培养24 h，采用ELISA检测各组白细胞介素-6（interleukin-6，IL-6）、IL-17、IL-1β、IL-8、核因子-κB（nuclear factor-κB，NF-κB）和肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平；qRT-PCR检测各组miR-21表达；Western blotting检测PTEN、B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）和磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）通路相关蛋白的表达。通过生物信息学预测miR-21与PTEN结合位点，并采用双荧光素酶报告基因检测系统验证CRCS干预后miR-21对PTEN的靶向作用。结果生物信息学共筛选出7个可能靶向PTEN的miRNA，其中党参干预后miR-21表达水平显著降低（P＜0.01），而miR-30a-3p、miR-198、miR-152-3p、miR-301a-3p、miR-320及miR-3691-5p表达变化均无统计学意义。CCK-8结果表明，2%、4%、6%、8% CRCS均可显著提高细胞活力（P＜0.01）。与模型组比较，CRCS和miR-21 inhibitor可显著上调细胞内PTEN和Bcl-2蛋白表达水平（P＜0.01），并降低miR-21、Bax、cleaved Caspase-3、p-PI3K/PI3K、p-Akt/Akt表达水平（P＜0.01）；与CRCS组比较，miR-21 mimics可显著削弱CRCS对PTEN和Bcl-2表达的上调作用（P＜0.01），以及对miR-21、Bax、cleaved Caspase-3、p-PI3K/PI3K、p-Akt/Akt表达的下调作用（P＜0.05、0.01）。双荧光素酶报告实验结果显示，在转染wt-Luc-PTEN质粒细胞中，CRCS干预后可显著提高荧光素酶活性（P＜0.01），miR-21 mimics可显著降低荧光素酶活性（P＜0.05），联合处理时，miR-21 mimics可部分削弱CRCS对荧光素酶活性升高的作用（P＜0.01）；在转染mut-Luc-PTEN质粒细胞中，CRCS干预及miR-21 mimics均未引起荧光素酶活性显著变化。结论 CRCS可通过抑制促炎因子释放及上皮细胞过度凋亡缓解UC，其机制至少部分依赖于miR-21/PTEN/PI3K/Akt轴。

关键词

党参 / 溃疡性结肠炎 / 上皮细胞过度凋亡 / miR-21/PTEN轴 / PI3K/Akt通路 / 党参炔苷

Abstract

收起

Objective To investigate the mechanism of Dangshen (Codonopsis Radix) containing serum (CRCS) on inflammatory factors release and epithelial cell excessive apoptosis in vitro model of ulcerative colitis (UC). Methods Bioinformatics analysis tools were used to predict and regulate the upstream miRNAs of phosphatase and tensin homolog (PTEN). qRT-PCR and Western blotting were used to detect the mRNA and protein expression levels of PTEN and the expressions of miR-21, miR-30a-3p, miR-198, miR-152-3p, miR-301a-3p, miR-320 and miR-3691-5p in colon tissues of UC rats model induced by enema 5% 2,4,6-trinitrobenzenesulfonic acid solution (TNBS)/ethanol composite solution. Lipopolysaccharide (LPS, 10 μg/mL) was used to induce human normal colon epithelial NCM460 cells to construct an in vitro model of UC, 1%, 2%, 4%, 6% and 8% CRCS intervention was administered for 24 h. The cell viability was detected by CCK-8 method. Subsequently, miR-21 mimics and miR-21 inhibitor were transfected into model cells, respectively, which were cultured in 6 % CRCS complete medium for 24 h. The levels of inflammatory factors interleukin-6 (IL-6), IL-17, IL-1β, IL-8, nuclear factor-κB (NF-κB) and tumor necrosis factor-α (TNF-α) in each group were detected by ELISA. The expression of miR-21 in each group was detected by qRT-PCR. Western blotting was used to detect the expressions of PTEN, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved cystein-asparate protease-3 (cleaved Caspase-3) and phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway related proteins. The binding sites of miR-21 and PTEN were predicted by bioinformatics, and the targeting effect of miR-21 on PTEN was verified by dual luciferase reporter gene detection system. Results A total of seven miRNAs that may target PTEN were screened by bioinformatics analysis. Among them, the expression of miR-21 was decreased significantly after CRCS intervention (P < 0.01), while the expressions of miR-30a-3p, miR-198, miR-152-3p, miR-301a-3p, miR-320 and miR-3691-5p had no statistical significance. The results of CCK-8 showed that 2%, 4%, 6% and 8% CRCS could significantly improve cell viability (P < 0.01). Compared with model group, CRCS and miR-21 inhibitor increased the expression levels of PTEN and Bcl-2 proteins in cells (P < 0.01), and decreased the expression levels of miR-21, Bax, cleaved Caspase-3, p-PI3K/PI3K and p-Akt/Akt (P < 0.01). Compared with CRCS group, miR-21 mimics significantly attenuated the up-regulation of PTEN and Bcl-2 expressions by CRCS (P < 0.01) and the down-regulation of miR-21, Bax, cleaved Caspase-3, p-PI3K/PI3K, p-Akt/Akt expressions (P < 0.05, 0.01). The results of dual luciferase reporter assay showed that in the transfected wt-Luc-PTEN plasmocytes, CRCS intervention could significantly increase luciferase activity (P < 0.01), and miR-21 mimics could significantly reduce luciferase activity (P < 0.01). In the combined treatment, miR-21 mimics could partially weaken the effect of CRCS on luciferase activity (P < 0.01). In transfected mut-Luc-PTEN plasmocytes, CRCS intervention and miR-21 mimics did not cause significant changes in luciferase activity. Conclusion CRCS could alleviate UC by inhibiting pro-inflammatory factors release and excessive apoptosis of epithelial cells. The mechanism is at least partially dependent on miR-21/PTEN/PI3K/Akt axis.

Key words

Codonopsis Radix / ulcerative colitis / epithelial cell excessive apoptosis / miR-21/PTEN axis / PI3K/Akt pathway / lobetyolin

引用本文

王春霞, 杨洁, 史奇, 李芳, 陈正君, 杨彦平, 冯永辉, 杨扶德. 党参含药血清通过调控miR-21/PTEN轴改善溃疡性结肠炎. 中草药, 2026 , 57 (5) : 1773 -1786 . DOI: 10.7501/j.issn.0253-2670.2026.05.016

WANG Chunxia, YANG Jie, SHI Qi, LI Fang, CHEN Zhengjun, YANG Yanping, FENG Yonghui, YANG Fude. Codonopsis Radix containing serum improves ulcerative colitis by regulating miR-21/PTEN axis[J]. Chinese Traditional and Herbal Drugs, 2026 , 57 (5) : 1773 -1786 . DOI: 10.7501/j.issn.0253-2670.2026.05.016

基金

收起

科技部国家重点研发计划项目 (2018YFC1706305); 甘肃省重点研发计划-社会发展类 (23YFFA0069); 甘肃省中医药研究中心开放课题 (zyzx-2023-10); 西北营养与环境相关疾病中医药防控协同创新中心2025年度开放基金课题 (ZYXT-25-04); 兰州市科技计划项目 (2024-9-78); 甘肃省优秀研究生“创新之星”项目 (2026CXZX-924)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

Ungaro R, Mehandru S, Allen P B, et al. Ulcerative colitis[J]. Lancet, 2017, 389(10080): 1756-1770.
Bressler B, Marshall J K, Bernstein C N, et al. Clinical practice guidelines for the medical management of nonhospitalized ulcerative colitis: The Toronto consensus[J]. Gastroenterology, 2015, 148(5): 1035-1058.e3.
Carbonnel F, Colombel J F, Filippi J, et al. Methotrexate is not superior to placebo for inducing steroid-free remission, but induces steroid-free clinical remission in a larger proportion of patients with ulcerative colitis[J]. Gastroenterology, 2016, 150(2): 380-388.
Järnerot G, Hertervig E, Friis-Liby I, et al. Infliximab as rescue therapy in severe to moderately severe ulcerative colitis: A randomized, placebo-controlled study[J]. Gastroenterology, 2005, 128(7): 1805-1811.
Shen B. Pouchitis: What every gastroenterologist needs to know[J]. Clin Gastroenterol Hepatol, 2013, 11(12): 1538-1549.
Øresland T, Bemelman W A, Sampietro G M, et al. European evidence based consensus on surgery for ulcerative colitis[J]. J Crohns Colitis, 2015, 9(1): 4-25.
Beyer-Berjot L, Maggiori L, Birnbaum D, et al. A total laparoscopic approach reduces the infertility rate after ileal pouch-anal anastomosis: A 2-center study[J]. Ann Surg, 2013, 258(2): 275-282.
Kani H T, Shen B. Male issues of the ileal pouch[J]. Inflamm Bowel Dis, 2015, 21(3): 716-722.
Zenlea T, Yee E U, Rosenberg L, et al. Histology grade is independently associated with relapse risk in patients with ulcerative colitis in clinical remission: A prospective study[J]. Am J Gastroenterol, 2016, 111(5): 685-690.
彭卓嵛, 陶丽芬, 蓝斯莹, 等. 基于数据挖掘对溃疡性结肠炎中医用药规律的分析[J]. 世界科学技术—中医药现代化, 2020, 22(7): 2269-2275.
Le Berre C, Honap S, Peyrin-Biroulet L. Ulcerative colitis[J]. Lancet, 2023, 402(10401): 571-584.
Patankar J V, Becker C. Cell death in the gut epithelium and implications for chronic inflammation[J]. Nat Rev Gastroenterol Hepatol, 2020, 17(9): 543-556.
Günther C, Neumann H, Neurath M F, et al. Apoptosis, necrosis and necroptosis: Cell death regulation in the intestinal epithelium[J]. Gut, 2013, 62(7): 1062-1071.
Maloy K J, Powrie F. Intestinal homeostasis and its breakdown in inflammatory bowel disease[J]. Nature, 2011, 474(7351): 298-306.
Cantley L C. The phosphoinositide 3-kinase pathway[J]. Science, 2002, 296(5573): 1655-1657.
Li Q Q, Li J, Yin L X, et al. Sophoricoside improved Crohn’s disease-like colitis by inhibiting intestinal epithelial cell apoptosis through PI3K/AKT signaling[J]. Int Immunopharmacol, 2024, 131: 111886.
Li F, Yang Y P, Ge J L, et al. Multi-omics revealed the mechanisms of Codonopsis pilosula aqueous extract in improving UC through blocking abnormal activation of PI3K/Akt signaling pathway[J]. J Ethnopharmacol, 2024, 319(Pt 2): 117220.
Bartel D P. Metazoan microRNAs[J]. Cell, 2018, 173(1): 20-51.
Cao Y Y, Wang Z H, Yan Y Q, et al. Enterotoxigenic bacteroidesfragilis promotes intestinal inflammation and malignancy by inhibiting exosome-packaged miR-149-3p[J]. Gastroenterology, 2021, 161(5): 1552-1566.e12.
Vermeire S, Nitcheu J, Gineste P, et al. Obefazimod in patients with moderate-to-severely active ulcerative colitis: Efficacy and safety analysis from the 96-week open-label maintenance phase 2b study[J]. J Crohns Colitis, 2025, 19(5): jjaf074.
Yu T X, Kalakonda S, Liu X Z, et al. Long noncoding RNA uc.230/CUG-binding protein 1 axis sustains intestinal epithelial homeostasis and response to tissue injury[J]. JCI Insight, 2022, 7(19): e156612.
Chen Y, Xiao Y, Ge W, et al. miR-200b inhibits TGF-β1-induced epithelial-mesenchymal transition and promotes growth of intestinal epithelial cells[J]. Cell Death Dis, 2013, 4(3): e541.
Maehama T, Dixon J E. The tumor suppressor, PTEN/ MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate[J]. J Biol Chem, 1998, 273(22): 13375-13378.
Lee Y R, Chen M, Pandolfi P P. The functions and regulation of the PTEN tumour suppressor: New modes and prospects[J]. Nat Rev Mol Cell Biol, 2018, 19(9): 547-562.
李芳, 陈正君, 葛俊李, 等. 党参经PI3K/Akt干预溃疡性结肠炎黏膜细胞铁死亡-线粒体动力学失衡的机制研究[J]. 中草药, 2023, 54(12): 3865-3877.
胡志航, 赵建斌, 柴建新, 等. 党参的毒理学试验研究[J]. 中国卫生检验杂志, 2018, 28(19): 2325-2329.
王领弟, 孙孟瑶, 张芳, 等. 体外细胞实验中药干预方法研究进展[J]. 中华中医药杂志, 2018, 33(4): 1448-1451.
孟雪, 吴鸿. 中药含药血清制备的研究进展与思考[J]. 辽宁中医杂志, 2025, 52(6): 122-124.
陈莉娟, 李彦龙, 杨维建, 等. 基于NLRP3/Caspase-1通路探讨薏苡仁多糖对溃疡性结肠炎体外炎症模型的干预作用及机制[J]. 重庆医科大学学报, 2023, 48(11): 1323-1330.
姜莎莎, 臧思源, 柳越冬, 等. 青黛对溃疡性结肠炎体外炎症模型的抗炎作用及机制[J]. 中华中医药杂志, 2021, 36(2): 1101-1104.
张惠琴, 杨海燕, 张红霞, 等. 党参标准汤剂指纹图谱及质量标准研究[J]. 中国现代应用药学, 2022, 39(22): 2921-2926.
彭锐, 马鹏, 莫让瑜, 等. 川党参药材质量标准研究[J]. 世界科学技术—中医药现代化, 2014, 16(3): 578-581.
Fisher K, Lin J M. microRNA in inflammatory bowel disease: Translational research and clinical implication[J]. World J Gastroenterol, 2015, 21(43): 12274-12282.
Endicott S J, Miller R A. PTEN activates chaperone-mediated autophagy to regulate metabolism[J]. Autophagy, 2024, 20(1): 216-217.
Miller K A, Degan S, Wang Y Q, et al. PTEN-regulated PI3K-p110 and AKT isoform plasticity controls metastatic prostate cancer progression[J]. Oncogene, 2024, 43(1): 22-34.
Salmena L, Carracedo A, Pandolfi P P. Tenets of PTEN tumor suppression[J]. Cell, 2008, 133(3): 403-414.
Alimonti A, Carracedo A, Clohessy J G, et al. Subtle variations in Pten dose determine cancer susceptibility[J]. Nat Genet, 2010, 42(5): 454-458.
Takagi T, Naito Y, Mizushima K, et al. Increased expression of microRNA in the inflamed colonic mucosa of patients with active ulcerative colitis[J]. J Gastroenterol Hepatol, 2010, 25(Suppl 1): S129-S133.
Casado-Bedmar M, Roy M, Berthet L, et al. Fecal let-7b and miR-21 directly modulate the intestinal microbiota, driving chronic inflammation[J]. Gut Microbes, 2024, 16(1): 2394249.
Ferragut Cardoso A P, Banerjee M, Nail A N, et al. miRNA dysregulation is an emerging modulator of genomic instability[J]. Semin Cancer Biol, 2021, 76: 120-131.
Qin T, Sun M K, Huang Y Y, et al. miR-221 on protective oxidative induced by selenium modified Codonopsis pilosula polysaccharide[J]. Int J Biol Macromol, 2024, 279(Pt 3): 134815.
Chen Y, Yu C, Hu F L, et al. Astragalus polysaccharides combined with Codonopsis pilosula polysaccharides modulates the physiological characteristics of trophoblasts via miR-92a-1-5p/CCR7 axis[J]. Tissue Cell, 2022, 77: 101827.
Devkota S, Wang Y W, Musch M W, et al. Dietary-fat-induced taurocholic acid promotes pathobiont expansion and colitis in Il10^−/− mice[J]. Nature, 2012, 487(7405): 104-108.

2026年第57卷第5期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.7501/j.issn.0253-2670.2026.05.016

接收时间：2025-11-18
首发时间：2026-09-09

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2025-11-18

基金

作者信息

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zcy/CN/10.7501/j.issn.0253-2670.2026.05.016

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT