Effects of cannabidiol on hepatic stellate cell activation and hepatic fibrosis induced by transforming growth factor beta1

doi:10.12307/2025.097

Abstract

Abstract: BACKGROUND: Cannabidiol has anti-inflammatory, antioxidant, and other pharmacological effects, and has no mental activity, so the research in liver disease is increasing day by day, but its effect on transforming growth factor-β1/Smad signal transduction pathway in hepatic stellate cells is not clear.
OBJECTIVE: To investigate the effect of cannabidiol on transforming growth factor-β1/Smad signal transduction pathway in hepatic stellate cells and its possible mechanism of anti-hepatic fibrosis.
METHODS: (1) In vitro experiment: Rat hepatic stellate cell line (HSC-T6) was selected and cultured in six groups. The control group was routinely cultured for 24 hours. The simple drug group was cultured with cannabidiol for 24 hours. The modeling group was cultured with transforming growth factor β1 for 24 hours. The modeling + low-dose drug group, the modeling + high-dose drug group, and the modeling + positive control group were cultured with transforming growth factor β1 for 24 hours, 1, 5 μmol/L cannabidiol and silymarin were cultured for 24 hours. After culture, the mRNA expression of α-smooth muscle actin and type I collagen, the levels of interleukin 1β and tumor necrosis factor α, and the protein expression of type I collagen and transforming growth factor β1/Smad signal transduction pathway were detected in each group. (2) In vivo experiments: C57BL/6J mice were randomly divided into five groups with eight mice in each group. Models were not established in the sham operation group. The liver fibrosis models were established by biliary ligation in the modeling group, the modeling+low-dose drug group, the modeling+high-dose drug group, and the modeling+positive control group. At 3 weeks after the modeling, 4, 8 mg/kg cannabidiol or silymarin were injected intraperitoneally, once a day, for 7 consecutive days. After administration, the liver function, liver pathological morphology, expression levels of α-smooth muscle actin, type I collagen, and transforming growth factor β1/Smad signal transduction pathway related protein were detected in each group.
RESULTS AND CONCLUSION: (1) In vitro experiment: Compared with the control group, mRNA expression of α-smooth muscle actin and type I collagen, interleukin 1β and tumor necrosis factor α, and protein expression of type I collagen, transforming growth factor β1 and p-Smad2/3 in HSC-T6 cells were increased (P < 0.05), while Smad7 protein expression was decreased (P < 0.05) in the modeling group. Two doses of cannabidiol could improve the above changes in HSC-T6 cells induced by transforming growth factor β1, and the improvement was more obvious in the modeling+high-dose drug group. (2) In vivo experiment: Compared with sham operation group, the activities of serum alanine aminotransferase and aspartate aminotransferase were increased (P < 0.05), inflammatory cell infiltration and collagen content in liver tissue were increased (P < 0.05), and the transforming growth factor β1/Smad signal transduction pathway was activated; α-smooth muscle actin and type I collagen expression levels were increased (P < 0.05) in the modeling group. Two doses of cannabidiol could reduce the changes of the above indexes in the modeling mice, and the effect was more obvious in the modeling+high-dose drug group. (3) It is indicated that cannabidiol inhibits hepatic fibrosis by suppressing the activation of transforming growth factor-β1/Smad signal transduction pathway in hepatic stellate cells.

Key words: cannabidiol, hepatic stellate cell, hepatic fibrosis, signal transduction pathway, transforming growth factor-β1/Smad, HSC-T6 cell

CLC Number:

Wang Lian, Xie Na, Zhao Peiling, Chen Hao, Li Duyou, Wang Yuping. Effects of cannabidiol on hepatic stellate cell activation and hepatic fibrosis induced by transforming growth factor beta1[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(23): 4965-4974.

Figures/Tables 4

2.1.3 各组HSC-T6细胞中α-平滑肌肌动蛋白、Ⅰ型胶原mRNA表达与对照组比较，单纯药物组HSC-T6细胞中纤维化因子α-平滑肌肌动蛋白、Ⅰ型胶原mRNA表达无明显变化(P > 0.05)，造模组HSC-T6细胞中Ⅰ型胶原与α-平滑肌肌动蛋白mRNA表达显著升高(P < 0.001)，说明转化生长因子β1能诱导HSC-T6细胞发生纤维化；与造模组比较，造模+低剂量药物组、造模+高剂量药物组、造模+阳性对照组HSC-T6细胞中α-平滑肌肌动蛋白、Ⅰ型胶原mRNA表达均降低(P < 0.001)；与造模+高剂量药物组比较，造模+低剂量药物组HSC-T6细胞中α-平滑肌肌动蛋白、Ⅰ型胶原mRNA表达降低(P < 0.05)，造模+阳性对照组HSC-T6细胞中α-平滑肌肌动蛋白、Ⅰ型胶原mRNA表达无明显变化(P > 0.05)，见图3。结果表明，大麻二酚对转化生长因子β1诱导的HSC-T6 细胞活化和纤维化基因的表达有抑制作用。 2.1.4 各组HSC-T6细胞中炎症因子表达的影响与对照组比较，单纯药物组HSC-T6细胞中白细胞介素1β和肿瘤坏死因子α水平无明显变化(P > 0.05)，造模组HSC-T6细胞中白细胞介素1β和肿瘤坏死因子α水平显著升高(P < 0.001)；与造模组比较，造模+低剂量药物组、造模+高剂量药物组、造模+阳性对照组HSC-T6细胞中白细胞介素1β和肿瘤坏死因子α水平降低(P < 0.05或P < 0.01或P < 0.001)；与造模+高剂量药物组比较，造模+低剂量药物组HSC-T6细胞中白细胞介素1β和肿瘤坏死因子α水平升高(P < 0.05)，造模+阳性对照组HSC-T6细胞中白细胞介素1β和肿瘤坏死因子α水平无明显变化(P > 0.05)，见图4。结果表明，大麻二酚对活化HSC-T6细胞的炎症反应具有抑制作用。 2.1.5 各组HSC-T6细胞中Ⅰ型胶原及转化生长因子β1/Smad信号转导通路相关蛋白的表达与对照组相比，单纯药物组HSC-T6 细胞中Ⅰ型胶原及转化生长因子β1/Smad信号转导通路相关蛋白表达水平无明显变化(P > 0.05)，造模组HSC-T6 细胞中Ⅰ型胶原、转化生长因子β1、p-Smad2/3 蛋白表达显著升高(P < 0.01或P < 0.001)，Smad7蛋白表达显著减少(P < 0.001)；与造模组比较，造模+低剂量药物组、造模+高剂量药物组、造模+阳性对照组HSC-T6细胞中Ⅰ型胶原、转化生长因子β1、p-Smad2/3 蛋白表达显著降低(P < 0.05或P < 0.01或P < 0.001)，Smad7蛋白表达升高(P < 0.05或P < 0.01或P < 0.001)；与造模+高剂量药物组比较，造模+低剂量药物组除Smad7外其余蛋白表达有明显变化(P < 0.05)，造模+阳性对照组除了Ⅰ型胶原与Smad7外其余蛋白表达有明显变化(P < 0.05)，见图5。这些结果表明，大麻二酚能抑制活化HSC-T6细胞中转化生长因子β1/Smad信号转导通路。"

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