Acupuncture inhibits mammalian target of rapamycin,promotes autophagy and attenuates neurological deficits in a rat model of hemorrhagic stroke

Abstract

Background:

Intracerebral hemorrhage (ICH) accounts for approximately 15% of all stroke cases. Previous studies suggested that acupuncture may improve ICH-induced neurological deficits. Therefore, we investigated the effects of acupuncture on neurological deficits in an animal model of ICH.

Methods:

Adult male Sprague–Dawley rats were injected with autologous blood (50 μL) into the right caudate nucleus. Additional rats underwent sham surgery as controls. ICH rats either received acupuncture (GV20 through GB7 on the side of the lesion) or sham acupuncture (1 cm to the right side of the traditional acupuncture point locations). Some ICH rats received acupuncture plus rapamycin injection into the right lateral ventricle. Neurological deficits in the various groups were assessed based on composite neurological score. The perihemorrhagic penumbra was analyzed by histopathology following hematoxylin–eosin staining. Levels of autophagy-related proteins light chain (LC)3 and p62 as well as of mammalian target of rapamycin (mTOR)-related proteins, and phosphorylated (p)-mTOR and p-S6K1 (ribosomal protein S6 kinase beta-1), were assessed by Western blotting.

Results:

Acupuncture significantly improved composite neurological scores 7 days after ICH (17.7 ± 1.49 vs 14.8 ± 1.32, p < 0.01). Acupuncture augmented autophagosome and autolysosome accumulation based on transmission electron microscopy. Acupuncture significantly increased expression of LC3 (p < 0.01) but decreased expression of p62 (p < 0.01). Acupuncture also reduced levels of p-mTOR and p-S6K1 (both p < 0.01).

Conclusion:

Acupuncture improved neurological deficits in a rat model of ICH, possibly by inhibiting the mTOR pathway and activating autophagy.

Keywords

acupuncture basic sciences complementary medicine neurology neurobiology stroke

Get full access to this article

View all access options for this article.

References

Kim

Yoon

BW.

Epidemiology, risk factors, and clinical features of intracerebral hemorrhage: an update. J Stroke 2017; 19(1): 3–10.

Van Asch

Luitse

Rinkel

, et al. Incidence, case fatality, and functional outcome of intracerebral haemorrhage over time, according to age, sex, and ethnic origin: a systematic review and meta-analysis. Lancet Neurol 2010; 9(2): 167–176.

Keep

Hua

Intracerebral haemorrhage: mechanisms of injury and therapeutic targets. Lancet Neurol 2012; 11(8): 720–731.

Kim

Bae

HJ.

Spontaneous intracerebral hemorrhage: management. J Stroke 2017; 19: 28–39.

Keep

Hoff

JT.

Mechanisms of brain injury after intracerebral haemorrhage. Lancet Neurol 2006; 5(1): 53–63.

Puyal

Ginet

Clarke

PG.

Multiple interacting cell death mechanisms in the mediation of excitotoxicity and ischemic brain damage: a challenge for neuroprotection. Prog Neurobiol 2013; 105: 24–48.

Klionsky

Abdelmohsen

Abe

, et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 2016; 12(1): 1–222.

Mehrpour

Esclatine

Beau

, et al. Autophagy in health and disease. 1. Regulation and significance of autophagy: an overview. Am J Physiol Cell Physiol 2010; 298: C776–C785.

Mizushima

Levine

Autophagy in mammalian development and differentiation. Nat Cell Biol 2010; 12(9): 823–830.

10.

Zhang

Zhong

, et al. miRNA-144 induces microglial autophagy and inflammation following intracerebral hemorrhage. Immunol Lett 2017; 182: 18–23.

11.

Duan

Wang

Feng

, et al. Roles of autophagy and endoplasmic reticulum stress in intracerebral hemorrhage-induced secondary brain injury in rats. CNS Neurosci Ther 2017; 23(7): 554–566.

12.

Liu

Xue

Zhang

, et al. Neuronal-targeted TFEB rescues dysfunction of the autophagy-lysosomal pathway and alleviates ischemic injury in permanent cerebral ischemia. Autophagy 2018; 10: 1–17.

13.

Zhang

Kai

, et al. Rapamycin prevents cerebral stroke by modulating apoptosis and autophagy in penumbra in rats. Ann Clin Transl Neurol 2017; 5: 138–146.

14.

Wan

Hua

, et al. Autophagy after experimental intracerebral hemorrhage. J Cereb Blood Flow Metab 2008; 28: 897–905.

15.

Harder

Bunkenborg

Andersen

JS.

Inducing autophagy: a comparative phosphoproteomic study of the cellular response to ammonia and rapamycin. Autophagy 2014; 10(2): 339–355.

16.

Uchida

Hotta

Acupuncture affects regional blood flow in various organs. Evid Based Complement Alternat Med 2008; 5(2): 145–151.

17.

Zou

Chen

Sun

, et al. Acupuncture inhibits Notch1 and Hes1 protein expression in the basal ganglia of rats with cerebral hemorrhage. Neural Regen Res 2015; 10(3): 457–462.

18.

Zhang

Dai

, et al. Baihui (DU20)-penetrating-Qubin (GB7) acupuncture inhibits apoptosis in the perihemorrhagic penumbra. Neural Regen Res 2018; 13: 1602–1608.

19.

Liu

Shang

Yang

, et al. Electroacupuncture protects against ischemic stroke by reducing autophagosome formation and inhibiting autophagy through the mTORC1-ULK1 complex-Beclin1 pathway. Int J Mol Med 2016; 37(2): 309–318.

20.

Shu

You

, et al. Electroacupuncture ameliorates cerebral ischemia-reperfusion injury by regulation of autophagy and apoptosis. Evid Based Complement Alternat Med 2016; 2016: 7297425.

21.

Ting

Jianbin

Luqi

Protective effect of electroacupuncture on neurons autophagy in perfusion period of cerebral ischemia. Neurosci Lett 2017; 20(661): 41–45.

22.

Zou

, et al. Electroacupuncture pretreatment induces tolerance against cerebral ischemia/reperfusion injury through inhibition of the autophagy pathway. Mol Med Rep 2015; 11(6): 4438–4446.

23.

Tian

Sun

, et al. Acupuncture promotes mTOR-independent autophagic clearance of aggregation-prone proteins in mouse brain. Sci Rep 2016; 21(6): 19714.

24.

Guo

Zhu

Tian

, et al. Electroacupuncture improves memory and protects neurons by regulation of the autophagy pathway in a rat model of Alzheimer’s disease. Acupunct Med 2016; 34(6): 449–456.

25.

Liu

Sun

Zou

, et al. Scalp acupuncture attenuates neurological deficits in a rat model of hemorrhagic stroke. Complement Ther Med 2017; 32: 85–90.

26.

George

Charles

. The rat brain in stereotaxic coordinates. 6th ed. Burlington, MA: Academic Press, pp. 101–102.

27.

Ashwal

Tone

Tian

, et al. Core and penumbral nitric oxide synthase activity during cerebral ischemia and reperfusion. Stroke 1998; 29(5): 1037–1046.

28.

Liu

, et al. MicroRNA-27b inhibition promotes Nrf2/ARE pathway activation and alleviates intracerebral hemorrhage-induced brain injury. Oncotarget 2017; 8: 70669–70684.

29.

Wang

Fei

, et al. Bone marrow mesenchymal stem cells ameliorate neurological deficits and blood-brain barrier dysfunction after intracerebral hemorrhage in spontaneously hypertensive rats. Int J Clin Exp Pathol 2015; 8(5): 4715–4724.

30.

Liu

Dai

Zou

, et al. Acupuncture through Baihui (DU20) to Qubin (GB7) mitigates neurological impairment after intracerebral hemorrhage. Neural Regen Res 2018; 13: 1425–1432.

31.

Niu

Dai

Zou

, et al. Autophagy, endoplasmic reticulum stress and the unfolded protein response in intracerebral hemorrhage. Transl Neurosci 2017; 8: 37–48.

32.

Feng

Zhang

Zhu

, et al. Ubiquitin ligase SYVN1/HRD1 facilitates degradation of the SERPINA1 Z variant/α-1-antitrypsin Z variant via SQSTM1/p62-dependent selective autophagy. Autophagy 2017; 13(4): 686–702.

33.

Thorburn

Autophagy and its effects: making sense of double-edged swords. PLOS Biol 2014; 12(10): e1001967.

34.

Sun

Wang

, et al. bFGF plays a neuroprotective role by suppressing excessive autophagy and apoptosis after transient global cerebral ischemia in rats. Cell Death Dis 2018; 9: 172.