铁皮石斛多糖通过上皮间质转化和Notch信号通路体外抗肝纤维化作用机制

doi:10.12092/j.issn.1009-2501.2026.03.005

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (3): 337-343.doi: 10.12092/j.issn.1009-2501.2026.03.005

铁皮石斛多糖通过上皮间质转化和Notch信号通路体外抗肝纤维化作用机制

白雅洁¹(), 杨柳青², 范钦¹, 刘旭玮¹, 何雨宸³, 成家茂¹^,*(), 陈海燕⁴^,*()

1. 大理大学医学部基础医学院人体解剖学教研室，大理 671003，云南
2. 邢台医学院临床医学系医学影像教研室，邢台 054000，河北
3. 大理大学医学部临床医学院，大理 671003，云南
4. 大理大学医学部临床医学院超声教研室/大理大学第一附属医院超声科，大理 671013，云南

收稿日期:2026-02-03 修回日期:2026-02-14 出版日期:2026-03-26 发布日期:2026-04-03
通讯作者: 成家茂,陈海燕 E-mail:15617711278@163.com;276625041@qq.com;316573230@qq.com
作者简介:白雅洁，女，在读硕士研究生，研究方向：从事中医药抗肝纤维化作用机制研究。E-mail：15617711278@163.com
基金资助:
2021年云南省地方本科高校基础研究联合专项资金面上项目（202101BA070001～101）；2023年大理护理职业学院校级科研课题项目（Dhzyxj2023009）；2024年云南省教育厅科学研究基金项目（2024J1822）；2025年云南省地方高校联合专项-青年项目（202501BA070001-024）

The anti-hepatic fibrosis mechanism of Dendrobium officinale poly saccharides in vitro through epithelial-mesenchymal transition and Notch signaling pathway

Yajie BAI¹(), Liuqing YANG², Qin FAN¹, Xuwei LIU¹, Yuchen HE³, Jiamao CHENG¹^,*(), Haiyan CHEN⁴^,*()

1. Department of Human Anatomy, School of Basic Medicine, Medical College of Dali University, Dali 671003, Yunnan, China
2. Department of Medical Imaging, Clinical Medicine Division, Xingtai Medical College, Xingtai 054000, Hebei, China
3. School of Clinical Medicine, Medical College of Dali University, Dali 671003, Yunnan, China
4. Department of Ultrasound, School of Clinical Medicine, Medical College of Dali University/Ultrasound Department of the First Affiliated Hospital of Dali University, Dali 671013, Yunnan, China

Received:2026-02-03 Revised:2026-02-14 Online:2026-03-26 Published:2026-04-03
Contact: Jiamao CHENG,Haiyan CHEN E-mail:15617711278@163.com;276625041@qq.com;316573230@qq.com

摘要/Abstract

摘要：

目的: 探讨铁皮石斛多糖（DOP）通过上皮间质转化（EMT）和Notch 信号通路对TGF-β1 诱导肝星状细胞（HSC）系HSC-T6 细胞活化的影响及其抗肝纤维化（HF）机制。方法: 采用CCK-8 法检测DOP 对HSC-T6 细胞的给药浓度，并计算24 h半数抑制浓度（IC₅₀），确定安全给药剂量。将HSC-T6细胞随机分为正常对照组，TGF-β1活化组（10 ng/mL）以及低、中、高剂量DOP组（1.6、3.2、6.4 mg/mL）。TGF-β1活化组及各药物组均加入10 ng/mL 的TGF-β1，各药物组同时给予相应浓度药物，共处理24 h。采用CCK-8法检测DOP干预后各组HSC-T6细胞的活性，采用RT-qPCR和Western blot法检测各组纤维化相关指标（α-SMA、COL-I）、EMT相关指标（E-cadherin、Vimentin、ZEB1）以及Notch信号通路相关指标（Notch1、Jagged1、Hes1）的mRNA及蛋白表达水平。结果: DOP干预24 h后，与TGF-β1活化组相比，中、高剂量DOP 组的HSC-T6细胞活性受到显著抑制（P<0.01），且细胞中α-SMA、COL-I、Vimentin、ZEB1以及Notch1、Jagged1、Hes1的mRNA及蛋白表达水平均显著降低，E-cadherin表达显著升高（P<0.05或0.01），而低剂量组对HSC-T6细胞的影响较小。结论: DOP可通过直接抑制HSC的活化以及TGF-β1活化HSC中的EMT、Notch信号通路，或间接通过抑制Notch信号抑制EMT，发挥抗HF作用。

关键词: 铁皮石斛多糖, 肝纤维化, 肝星状细胞, 上皮-间质转化, Notch信号通路

Abstract:

AIM: To explore the effect of Dendrobium officinale polysaccharide (DOP) on the activation of rat hepatic stellate cells (HSC-T6) induced by TGF-β1, and its anti-hepatic fibrosis (HF) mechanism, through epithelial-mesenchymal transition (EMT) and Notch signaling pathway. METHODS: The administration concentration of DOP to HSC-T6 cells was detected, and its 24-hour half maximal inhibitory concentration (IC₅₀) was calculated to determine the safe dosage for administration by CCK-8 method. HSC-T6 cells were randomly divided into normal group, TGF-β1 group (10 ng/mL), and the low, medium, and high DOP groups (1.6, 3.2, 6.4 mg/mL, respectively). TGF-β1 at a concentration of 10 ng/mL was added into the TGF-β1 group and each drug group, and the corresponding concentrations of DOP were administered to different drug groups. After 24 hours culture, the activity of HSC-T6 cells in each group after was detected by CCK-8 method, and the mRNA and protein expression levels of fibrosis indicators (α-SMA, COL-I), EMT indicators (E-cadherin, Vimentin, ZEB1), and Notch signaling pathway indicators (Notch1, Jagged1, Hes1) were detected by RT-qPCR and Western blot methods. RESULTS: After 24 hours of DOP intervention, compared with the TGF-β1 group, the activity of HSC-T6 cells was significantly inhibited (P<0.01), and the expression levels of α-SMA, COL-I, Vimentin, ZEB1, Notch1, Jagged1, Hes1 mRNAs and proteins in the cells were significantly reduced, and the expression of E-cadherin mRNA and protein was significantly increased (P<0.05 or 0.01) in the medium and high DOP groups, while there is a smaller effect on HSC-T6 cells in the low DOP group. CONCLUSION: DOP can exert anti-HF effects by directly inhibiting the activation of HSCs, EMT and Notch signaling pathways in the TGF-β1 activated HSCs, or indirectly by inhibiting the Notch signaling pathway to suppress EMT.

Key words: Dendrobium officinale polysaccharides, hepatic fibrosis, hepatic stellate cell, epithelial-mesenchymal transition, Notch signaling pathway

中图分类号:

R965.2

白雅洁, 杨柳青, 范钦, 刘旭玮, 何雨宸, 成家茂, 陈海燕. 铁皮石斛多糖通过上皮间质转化和Notch信号通路体外抗肝纤维化作用机制[J]. 中国临床药理学与治疗学, 2026, 31(3): 337-343.

Yajie BAI, Liuqing YANG, Qin FAN, Xuwei LIU, Yuchen HE, Jiamao CHENG, Haiyan CHEN. The anti-hepatic fibrosis mechanism of Dendrobium officinale poly saccharides in vitro through epithelial-mesenchymal transition and Notch signaling pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(3): 337-343.

图/表 4

表 1

Table 1 Primer sequences and their product lengths

Primer name	Primer sequence 5'-3'	Product length (bp)
α-SMA	F:5′-ACCATCGGGAATGAACGCTT-3′ R:5′-CTGTCAGCAATGCCTGGGTA-3′	191
Col-I	F:5′-CCCAGCGGTGGTTATGACTT-3′ R: 5′-TCGATCCAGTACTCTCCGCT-3′	257
E-cadherin	F: 5′-CTGGGGTCATTTCCACTCGG-3′ R: 5′-GTGGCAATGGGTGAACCATCA-3′	112
Vimentin	F: 5′-CGCCAGATGCGTGAAATG-3′ F:5′-CCAGAGGAAGTGACTCCAGGTTA-3′	277
ZEB1	F:5′-CGCGGCGCAATAACGTTACAAAT-3′ R: 5′-GCTCATCATCTGGCACACCA-3′	141
Notch1^[9]	F: 5′-TGGATGAGGAAGACAAGCATTA-3′ R: 5′-GAAAAGCCACCGAGATAGTCAG-3′	136
Jagged1^[10]	F: 5′-GAGACCTCCTCGGGCTTTGA-3′ R: 5′-GCACACGCACTTGAATCCAT-3′	144
Hes1^[10]	F: 5′-CAAGCTGGAGAAGGCAGACAT-3′ R: 5′-CCTCGTTCATGCACTCGCTG-3′	143
β-actin	F: 5′-TGCTATGTTGCCCTAGACTTCG-3′ R: 5′-GTTGGCATAGAGGTCTTTACGG-3′	240

图 1

Fig.1 Cell viability assay of HSC-T6 ($\bar x $±s, n=3) Panel A shows the effect of TGF-β1 on the viability of HSC-T6 cells; panel B shows the effect of DOP on the viability of HSC-T6 cells; and panel C shows the effect of DOP on the viability of TGF-β1-activated HSC-T6 cells; NG: normal control group; AG: activated group; LDG: low-dose group; MDG: medium-dose group; HDG: high-dose group; bP<0.05, cP<0.01, compared with 0 ng/mL; fP<0.01, compared with normal control group; iP<0.01, compared with the model group; lP<0.01, compared with the low-dose group; oP<0.01, compared with the medium-dose group.

图 2

Fig.2 The mRNA and protein expression of HF and EMT related indicators in each group of rats ($\bar x $±s, n=3) NG: normal control group; AG: activated group; LDG: low-dose DOP group; MDG: medium-dose DOP group; HDG: high-dose DOP group; A-E: mRNA expression levels of α-SMA, COL-I, E-cadherin, Vimentin, and ZEB1. F-J: protein expression levels of α-SMA, COL-I, E-cadherin, Vimentin, and ZEB1; bP<0.05, cP<0.01, compared with normal control group; eP<0.05, fP<0.01, compared with the model group.

图 3

Fig.3 Expression of Notch signaling pathway-related factors in each group of rats ($\bar x $±s, n=3) NG: normal control group; AG: activated group; LDG: low-dose DOP group; MDG: medium-dose DOP group; HDG: high-dose DOP group; A-C: mRNA expression levels of Notch1, Jagged1 and Hes1; D-F: protein expression levels of Notch1, Jagged1 and Hes1; cP<0.01, compared with normal control group; eP<0.05, fP<0.01, compared with the model group.

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铁皮石斛多糖通过上皮间质转化和Notch信号通路体外抗肝纤维化作用机制

The anti-hepatic fibrosis mechanism of Dendrobium officinale poly saccharides in vitro through epithelial-mesenchymal transition and Notch signaling pathway

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