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Rapamycin Alleviates Hypertriglyceridemia-Related Acute Pancreatitis via Restoring Autophagy Flux and Inhibiting Endoplasmic Reticulum Stress

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Abstract

Hypertriglyceridemia (HTG) can aggravate acute pancreatitis (AP), but its pathogenesis remains unclear. As autophagic activity is closely related to lipid metabolism and AP, we investigated the autophagic response in models of AP aggravated by HTG and explored whether rapamycin has a protective effect against HTG-related pancreatitis. HTG-associated AP models were established in vivo in rats and in vitro. The degree of inflammation, pancreatic injury, the expression of endoplasmic reticulum (ER) stress, and autophagy markers (P62, LC3) were compared. Autophagic flux were assessed using immunostaining, electron microscopy, and immunoblotting. Compared with the normal diet group, the high-fat diet (HFD) AP group exhibited more severe pancreatic injury, apoptosis, and blocked autophagic flux. In addition, the three branches (PERK–eIF2α, ATF-6–GRP78, and IRE1–sXBP1) of the unfolded protein response and mTORC1/S6K1 pathway were activated in HFD AP models. Moreover, the same phenomena were confirmed in vitro in palmitic acid–stimulated pancreatic acinar cells. Preincubation with the mTOR inhibitor rapamycin restored the autophagic flux and markedly reduced the adverse effects of HTG. In conclusion, the autophagic flux is impaired in HFD-induced AP models and is strongly associated with ER stress. Rapamycin could prevent the aggravation of HTG-associated AP via inhibiting mTORC1/S6K1 pathway.

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Abbreviations

AP:

acute pancreatitis

ND:

normal diet

HFD:

high-fat diet

PA:

palmitic acid

ERS:

endoplasmic reticulum stress

HTG:

hypertriglyceridemia

UPR:

unfolded protein response

PACs:

pancreatic acinar cells

TG:

triglycerides

TC:

total cholesterol

CQ:

chloroquine

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Funding

This work was supported by grants from the Natural Science Foundation of China (No. 81970555),the Natural Science Foundation for Young Scholars of China (No. 8160030431), the Medical-engineering Cross Project of Shanghai JiaoTong University (YG2015MS29 and YG2014ZD10), and the Clinical Cultivation Research Project of Shanghai Shenkang Hospital Development Center (SHDC12017X09).

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Authors and Affiliations

  1. Shanghai Key Laboratory of Pancreatic Disease, Shanghai JiaoTong University School of Medicine, Shanghai, 201600, China

    Qixiang Mei, Yue Zeng, Chunlan Huang, Junyuan Zheng, Yuecheng Guo, Junjie Fan, Xinyuan Fu, Xingpeng Wang & Yingying Lu

  2. Department of Gastroenterology, Shanghai General Hospital, Shanghai, 201600, China

    Xingpeng Wang & Yingying Lu

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  1. Qixiang Mei
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Contributions

Y.Z designed the research plan, and Q.X.M performed the research. C.L.H, J.Y.Z, X.Y.F, and Y.C.G assisted in the animal experiments. J.J.F and J.Y.Z interpreted the results. Q.X.M and Y.Y.L generated the draft of the manuscript. X.P.W and Y.Y.L edited and revised the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Xingpeng Wang or Yingying Lu.

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Electronic Supplementary Material

ESM 1.

Supplement 1 (a) Flow chart of pancreatitis modeling (b-c) Plasma levels of TG and TC in the normal diet and HFD groups. (d-f) Serum levels of inflammatory cytokines (IL-1β, TNFα and IL-6) in rats 3, 6, 9, and 12 h after the initial caerulein injection. The data are provided as the mean ± SEM (n = 6 per group).*p < 0.05 versus nor-AP group, #p < 0.05 versus normal-diet AP group. (PNG 2189 kb)

High Resolution (TIF 714 kb)

ESM 2.

Supplement 2 (a-b) Representative anti-LC3 immunostaining in pancreatic tissue obtained from rats 9 h after the AP induction. Scale bar = 50 μm. (c-d) Expression of P62, LC3 in the pancreatic tissues by Western Blot at 9 h after ANP induction with or without chloroquine(CQ) in both normal diet and HFD groups. The data are provided as the mean ± SEM (n = 6 per group).*p < 0.05 versus nor-AP or nor-CQ group, #p < 0.05 versus normal-diet AP group. (PNG 1961 kb)

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Mei, Q., Zeng, Y., Huang, C. et al. Rapamycin Alleviates Hypertriglyceridemia-Related Acute Pancreatitis via Restoring Autophagy Flux and Inhibiting Endoplasmic Reticulum Stress. Inflammation 43, 1510–1523 (2020). https://doi.org/10.1007/s10753-020-01228-7

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