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

doi:10.12092/j.issn.1009-2501.2026.03.005

Abstract

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

CLC Number:

R965.2

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.

Figures/Tables 4

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

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.

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.

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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