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Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss. Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathway

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Abstract

Objective

To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis (L.) Juss. (EDC) in human hepatic stellate cells (HSCs) in vitro and in a carbon tetrachloride (CCl4)-induced hepatic fibrosis mouse model in vivo.

Methods

For in vitro study, HSCs were pre-treated with platelet-derived growth factor (10 ng/mL) for 2 h to ensure activation and treated with EDC for 24 h and 48 h, respectively. The effect of EDC on HSCs was assessed using cell counting kit-8 assay, EdU staining, transmission electron microscopy, immunofluorescence staining, and Western blot, respectively. For in vivo experiments, mice were intraperitoneally injected with CCl4 (2 ° L/g, adjusted to a 25% concentration in olive oil), 3 times per week for 6 weeks, to develop a hepatic fibrosis model. Forty 8-week-old male C57BL/6 mice were divided into 4 groups using a random number table (n=10), including control, model, positive control and EDC treatment groups. Mice in the EDC and colchicine groups were intragastrically administered EDC (0.5 g/kg) or colchicine (0.2 mg/kg) once per day for 6 weeks. Mice in the control and model groups received an equal volume of saline. Biochemical assays and histological examinations were used to assess liver damage. Protein expression levels of α -smooth muscle actin (α -SMA) and microtubule-associated protein light chain 3B (LC3B) were measured by Western blot.

Results

EDC reduced pathological damage associated with liver fibrosis, downregulated the expression of α -SMA and upregulated the expression of LC3B (P<0.05), both in HSCs and the CCl4-induced liver fibrosis mouse model. The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhanced α -SMA protein expression levels (P<0.01). The results also found that the levels of phosphoinositide (PI3K), p-PI3K, AKT, p-AKT, mammalian target of rapamycin (mTOR), p-mTOR, and p-p70S6K all decreased after EDC treatment (P<0.05).

Conclusions

EDC has anti-hepatic fibrosis activity by inducing autophagy and might be a potential drug to be further developed for human liver fibrosis therapy.

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Authors and Affiliations

  1. College of Integrated Traditional Chinese and Western Medicine, Jining Medical University, Jining, Shandong Province, 272000, China

    Yuan Liu, Yan-meng Bi & Bing Sun

  2. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China

    Yuan Liu, Ting Pan, Ting Zeng, Chan Mo, Lei Gao & Zhi-ping Lyu

  3. Department of Traditional Chinese Medicine, Affiliated Hospital of Jining Medical University, Jining, Shandong Province, 272000, China

    Yan-meng Bi

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  1. Yuan Liu
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  4. Ting Zeng
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  6. Bing Sun
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Correspondence to Zhi-ping Lyu.

Additional information

Supported by the National Natural Science Foundation of China (No. 81673774) and the Administration of Traditional Chinese Medicine of Shandong Province (2019-0447)

Conflict of Interest

The authors declare that there is no conflict of interest.

Author Contributions

Lyu ZP, Gao L and Sun B participated in conception and design of the research. Liu Y and Bi YM carried out the experiments. Liu Y and Pan T drafted the manuscript. Zeng T participated in revision of the manuscript. Mo C participated in statistical analysis. All authors approved the final manuscript for publication.

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Liu, Y., Bi, Ym., Pan, T. et al. Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss. Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathway. Chin. J. Integr. Med. 28, 60–68 (2022). https://doi.org/10.1007/s11655-021-3298-5

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  • DOI: https://doi.org/10.1007/s11655-021-3298-5

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