Skip to main content

Ginsenoside Rd Is Efficacious Against Acute Ischemic Stroke by Suppressing Microglial Proteasome-Mediated Inflammation

Abstract

A great deal of attention has been paid to neuroprotective therapies for cerebral ischemic stroke. Our two recent clinical trials showed that ginsenoside Rd (Rd), a kind of monomeric compound extracted from Chinese herbs, Panax ginseng and Panax notoginseng, was safe and efficacious for the treatment of ischemic stroke. In this study, we conducted a pooled analysis of the data from 199 patients with acute ischemic stroke in the first trial and 390 in the second to reanalyze the efficacy and safety of Rd. Moreover, animal stroke models were carried out to explore the possible molecular mechanisms underlying Rd neuroprotection. The pooled analysis showed that compared with placebo group, Rd could improve patients’ disability as assessed by modified Rankin Scale (mRS) score on day 90 post-stroke and reduce neurologic deficits on day 15 or day 90 post-stroke as assessed by NIH Stroke Scale (NIHSS) and Barthel Index (BI) scores. For neuroprotective mechanisms, administration of Rd 4 h after stroke could inhibit ischemia-induced microglial activation, decrease the expression levels of various proinflammatory cytokines, and suppress nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor, alpha (IκBα) phosphorylation and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) nuclear translocation. An in vitro proteasome activity assay revealed a significant inhibitory effect of Rd on proteasome activity in microglia. Interestingly, Rd was showed to have less side effects than glucocorticoid. Therefore, our study demonstrated that Rd could safely improve the outcome of patients with ischemic stroke, and this therapeutic effect may result from its capability of suppressing microglial proteasome activity and sequential inflammation.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. 1.

    Feigin VL, Lawes CM, Bennett DA, Barker-Collo SL, Parag V (2009) Worldwide stroke incidence and early case fatality reported in 56 population-based studies: a systematic review. Lancet Neurol 8(4):355–369. doi:10.1016/S1474-4422(09)70025-0

    Article  PubMed  Google Scholar 

  2. 2.

    Norrving B, Kissela B (2013) The global burden of stroke and need for a continuum of care. Neurology 80(3 Suppl 2):S5–S12. doi:10.1212/WNL.0b013e3182762397

    Article  PubMed  Google Scholar 

  3. 3.

    Sahota P, Savitz SI (2011) Investigational therapies for ischemic stroke: neuroprotection and neurorecovery. Neurotherapeutics 8(3):434–451. doi:10.1007/s13311-011-0040-6

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Donnan GA, Davis SM, Parsons MW, Ma H, Dewey HM, Howells DW (2011) How to make better use of thrombolytic therapy in acute ischemic stroke. Nat Rev Neurol 7(7):400–409. doi:10.1038/nrneurol.2011.89

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Wardlaw JM, Murray V, Berge E, del Zoppo G, Sandercock P, Lindley RL, Cohen G (2012) Recombinant tissue plasminogen activator for acute ischaemic stroke: an updated systematic review and meta-analysis. Lancet 379(9834):2364–2372. doi:10.1016/S0140-6736(12)60738-7

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Stroke Unit Trialists C (2007) Organised inpatient (stroke unit) care for stroke. Cochrane Database Syst Rev 4, CD000197. doi:10.1002/14651858.CD000197.pub2

    Google Scholar 

  7. 7.

    Sutherland BA, Minnerup J, Balami JS, Arba F, Buchan AM, Kleinschnitz C (2012) Neuroprotection for ischaemic stroke: translation from the bench to the bedside. Int J Stroke 7(5):407–418. doi:10.1111/j.1747-4949.2012.00770.x

    Article  PubMed  Google Scholar 

  8. 8.

    Wahlgren NG, Ahmed N (2004) Neuroprotection in cerebral ischaemia: facts and fancies—the need for new approaches. Cerebrovasc Dis 17(Suppl 1):153–166. doi:10.1159/000074808

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Edaravone Acute Infarction Study G (2003) Effect of a novel free radical scavenger, edaravone (MCI-186), on acute brain infarction. Randomized, placebo-controlled, double-blind study at multicenters. Cerebrovasc Dis 15(3):222–229

    Article  Google Scholar 

  10. 10.

    Hill MD, Martin RH, Mikulis D, Wong JH, Silver FL, Terbrugge KG, Milot G, Clark WM et al (2012) Safety and efficacy of NA-1 in patients with iatrogenic stroke after endovascular aneurysm repair (ENACT): a phase 2, randomised, double-blind, placebo-controlled trial. Lancet Neurol 11(11):942–950. doi:10.1016/S1474-4422(12)70225-9

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Liu X, Wang L, Wen A, Yang J, Yan Y, Song Y, Ren H, Wu Y et al (2012) Ginsenoside-Rd improves outcome of acute ischaemic stroke—a randomized, double-blind, placebo-controlled, multicenter trial. Eur J Neurol Offic J Eur Fed Neurol Soc 19(6):855–863. doi:10.1111/j.1468-1331.2011.03634.x

    CAS  Google Scholar 

  12. 12.

    Liu X, Xia J, Wang L, Song Y, Yang J, Yan Y, Ren H, Zhao G (2009) Efficacy and safety of ginsenoside-Rd for acute ischaemic stroke: a randomized, double-blind, placebo-controlled, phase II multicenter trial. Eur J Neurol Offic J Eur Fed Neurol Soc 16(5):569–575. doi:10.1111/j.1468-1331.2009.02534.x

    CAS  Google Scholar 

  13. 13.

    Li L, Liu J, Yan X, Qin K, Shi M, Lin T, Zhu Y, Kang T et al (2011) Protective effects of ginsenoside Rd against okadaic acid-induced neurotoxicity in vivo and in vitro. J Ethnopharmacol 138(1):135–141. doi:10.1016/j.jep.2011.08.068

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Ye R, Han J, Kong X, Zhao L, Cao R, Rao Z, Zhao G (2008) Protective effects of ginsenoside Rd on PC12 cells against hydrogen peroxide. Biol Pharm Bull 31(10):1923–1927

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Ye R, Kong X, Yang Q, Zhang Y, Han J, Li P, Xiong L, Zhao G (2011) Ginsenoside rd in experimental stroke: superior neuroprotective efficacy with a wide therapeutic window. Neurotherapeutics 8(3):515–525. doi:10.1007/s13311-011-0051-3

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Ye R, Kong X, Yang Q, Zhang Y, Han J, Zhao G (2011) Ginsenoside Rd attenuates redox imbalance and improves stroke outcome after focal cerebral ischemia in aged mice. Neuropharmacology 61(4):815–824. doi:10.1016/j.neuropharm.2011.05.029

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Ye R, Li N, Han J, Kong X, Cao R, Rao Z, Zhao G (2009) Neuroprotective effects of ginsenoside Rd against oxygen-glucose deprivation in cultured hippocampal neurons. Neurosci Res 64(3):306–310. doi:10.1016/j.neures.2009.03.016

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Ye R, Yang Q, Kong X, Han J, Zhang X, Zhang Y, Li P, Liu J et al (2011) Ginsenoside Rd attenuates early oxidative damage and sequential inflammatory response after transient focal ischemia in rats. Neurochem Int 58(3):391–398. doi:10.1016/j.neuint.2010.12.015

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Ye R, Zhang X, Kong X, Han J, Yang Q, Zhang Y, Chen Y, Li P et al (2011) Ginsenoside Rd attenuates mitochondrial dysfunction and sequential apoptosis after transient focal ischemia. Neuroscience 178:169–180. doi:10.1016/j.neuroscience.2011.01.007

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Zhang C, Du F, Shi M, Ye R, Cheng H, Han J, Ma L, Cao R et al (2012) Ginsenoside Rd protects neurons against glutamate-induced excitotoxicity by inhibiting ca(2+) influx. Cell Mol Neurobiol 32(1):121–128. doi:10.1007/s10571-011-9742-x

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Chamorro A, Hallenbeck J (2006) The harms and benefits of inflammatory and immune responses in vascular disease. Stroke 37(2):291–293. doi:10.1161/01.STR.0000200561.69611.f8

    Article  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Samson Y, Lapergue B, Hosseini H (2005) Inflammation and ischaemic stroke: current status and future perspectives. Rev Neurol (Paris) 161(12 Pt 1):1177–1182

    CAS  Article  Google Scholar 

  23. 23.

    Trendelenburg G (2008) Acute neurodegeneration and the inflammasome: central processor for danger signals and the inflammatory response? J Cereb Blood Flow Metab 28(5):867–881. doi:10.1038/sj.jcbfm.9600609

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Iadecola C, Alexander M (2001) Cerebral ischemia and inflammation. Curr Opin Neurol 14(1):89–94

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Zhang YX, Wang L, Xiao EL, Li SJ, Chen JJ, Gao B, Min GN, Wang ZP et al (2013) Ginsenoside-Rd exhibits anti-inflammatory activities through elevation of antioxidant enzyme activities and inhibition of JNK and ERK activation in vivo. Int Immunopharmacol 17(4):1094–1100

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Zhang Y, Zhou L, Zhang X, Bai J, Shi M, Zhao G (2012) Ginsenoside-Rd attenuates TRPM7 and ASIC1a but promotes ASIC2a expression in rats after focal cerebral ischemia. Neurological Sci Offic J Ital Neurol Soc Ital Soc Clin Neurophysiol 33(5):1125–1131. doi:10.1007/s10072-011-0916-6

    CAS  Google Scholar 

  27. 27.

    Yenari MA, Giffard RG (2001) Ischemic vulnerability of primary murine microglial cultures. Neurosci Lett 298(1):5–8

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Yenari MA, Kauppinen TM, Swanson RA (2010) Microglial activation in stroke: therapeutic targets. Neurotherapeutics 7(4):378–391. doi:10.1016/j.nurt.2010.07.005

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Dixit V, Mak TW (2002) NF-kappaB signaling. Many roads lead to madrid. Cell 111(5):615–619

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Chen J, Chen ZJ (2013) Regulation of NF-kappaB by ubiquitination. Curr Opin Immunol 25(1):4–12. doi:10.1016/j.coi.2012.12.005

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Hayden MS, Ghosh S (2008) Shared principles in NF-kappaB signaling. Cell 132(3):344–362. doi:10.1016/j.cell.2008.01.020

    CAS  Article  PubMed  Google Scholar 

  32. 32.

    Quinn TJ, Dawson J, Walters MR, Lees KR (2009) Reliability of the modified Rankin Scale: a systematic review. Stroke 40(10):3393–3395. doi:10.1161/STROKEAHA.109.557256

    Article  PubMed  Google Scholar 

  33. 33.

    Lees KR, Bath PM, Schellinger PD, Kerr DM, Fulton R, Hacke W, Matchar D, Sehra R et al (2012) Contemporary outcome measures in acute stroke research: choice of primary outcome measure. Stroke 43(4):1163–1170. doi:10.1161/STROKEAHA.111.641423

    Article  PubMed  Google Scholar 

  34. 34.

    Saver JL (2011) Optimal end points for acute stroke therapy trials: best ways to measure treatment effects of drugs and devices. Stroke 42(8):2356–2362. doi:10.1161/STROKEAHA.111.619122

    Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Chen YF (2012) Traditional Chinese herbal medicine and cerebral ischemia. Front Biosci (Elite Ed) 4:809–817

    Article  Google Scholar 

  36. 36.

    Kaste M (2012) Is the door open again for neuroprotection trials in stroke? Lancet Neurol 11(11):930–931. doi:10.1016/S1474-4422(12)70229-6

    Article  PubMed  Google Scholar 

  37. 37.

    Kaste M (2012) Is there a future for neuroprotective agents in acute ischaemic stroke? Eur J Neurol Offic J Eur Fed Neurol Soc 19(6):797–798. doi:10.1111/j.1468-1331.2011.03633.x

    CAS  Google Scholar 

  38. 38.

    Muir KW, Tyrrell P, Sattar N, Warburton E (2007) Inflammation and ischaemic stroke. Curr Opin Neurol 20(3):334–342. doi:10.1097/WCO.0b013e32813ba151

    CAS  Article  PubMed  Google Scholar 

  39. 39.

    Wang L, Zhang Y, Wang Z, Li S, Min G, Wang L, Chen J, Cheng J et al (2012) Inhibitory effect of ginsenoside-Rd on carrageenan-induced inflammation in rats. Can J Physiol Pharmacol 90(2):229–236. doi:10.1139/y11-127

    CAS  Article  PubMed  Google Scholar 

  40. 40.

    Yang XL, Guo TK, Wang YH, Huang YH, Liu X, Wang XX, Li W, Zhao X et al (2012) Ginsenoside Rd attenuates the inflammatory response via modulating p38 and JNK signaling pathways in rats with TNBS-induced relapsing colitis. Int Immunopharmacol 12(2):408–414. doi:10.1016/j.intimp.2011.12.014

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 31170801 to S.M., No. 81372115 to Z.G.Y., No. 81171236 to L.X.D., and No. 81371365 to Z.G.).

Conflict of Interest

The authors declare no conflict of interests.

Author information

Affiliations

Authors

Corresponding authors

Correspondence to Xuedong Liu, Gang Zhao or Ming Shi.

Additional information

Guangyun Zhang, Feng Xia, and Yunxia Zhang contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Figure 1
figure7

Schematic diagrams for experimental procedures of MCAO in rats (A) and cultured microglia subjected to OGD or LPS injury (B). (GIF 21 kb)

Supplementary Figure 2
figure8

Disposition of patients in the pooled study. (GIF 26 kb)

Supplementary Figure 3
figure9

Rd inhibits p65 nuclear translocation. Immunofluorescence staining of NF-κB p65 and Hoechst 33342 (Ho) in rat cortical penumbra regions in Sham (A-C), MCAO (D-F), and MCAO+Rd (G-I) groups. Arrows and arrowheads show cytosolic and nuclear location of p65 immunoreactivities, respectively. Scale bar: 10 μm. (GIF 412 kb)

ESM 1

(DOCX 18 kb)

High resolution image (TIFF 253 kb)

High resolution image (TIFF 529 kb)

High resolution image (TIFF 4820 kb)

Supplemental Table 1

Baseline characteristics of patients with ischemic stroke included in the full analysis set (FAS) (DOCX 14 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, G., Xia, F., Zhang, Y. et al. Ginsenoside Rd Is Efficacious Against Acute Ischemic Stroke by Suppressing Microglial Proteasome-Mediated Inflammation. Mol Neurobiol 53, 2529–2540 (2016). https://doi.org/10.1007/s12035-015-9261-8

Download citation

Keywords

  • Ginsenoside Rd
  • Ischemic stroke
  • Pooled analysis
  • Microglia
  • Proteasome
  • Inflammation