Tong Luo Jiu Nao injection, a traditional Chinese medicinal preparation, inhibits MIP-1β expression in brain microvascular endothelial cells injured by oxygen-glucose deprivation

J Ethnopharmacol. 2012 May 7;141(1):151-7. doi: 10.1016/j.jep.2012.02.013. Epub 2012 Feb 17.

Abstract

Ethnopharmacological relevance: Tong Luo Jiu Nao injection (TLJN), a modern Chinese formula based on Traditional Chinese Medicine theory, has been used to treat ischemic stroke and vascular dementia. TLJN belongs to the ethnopharmacological family of medicines.

Aim of the study: To investigate the protective effect of TLJN on oxygen-glucose deprivation (OGD) induced-injury of brain microvascular endothelial cells (BMECs).

Materials and methods: The model of OGD was established in the primarily cultured BMECs. TLJN was added to the OGD-injured BMECs for 6h. A series of assays were used to detect the effects of TLJN on: (i) MIP-1β content in BMECs conditioned media (CM) by ELISA; (ii) MIP-1β protein expression in BMECs by western blotting and immunocytochemistry; (iii) the expression of CCR5, receptor of MIP-1β, in BMECs by western blotting; (iv) the proliferative activity of microglial cells via the Cell Counting Kit-8 (CCK-8).

Results: Our results showed that the OGD-injured BMECs presented with large amounts of secretion and expression of MIP-1β and up-regulation of CCR5. Also, the CM of OGD-injured BMECs remarkably increased the proliferative activity of microglial cells. The TLJN-treated BMECs exhibited a reduction of MIP-1β content in BMECs-CM and a down-regulation of MIP-1β and CCR5 expression. In addition, an inhibitory effect of CM of OGD-injured plus TLJN injection-treated BMECs on microglial proliferation was also found.

Conclusion: TLJN reduced the expression of MIP-1β and CCR5 in OGD-injured BMECs, and the CM of OGD-injured plus TLJN injection-treated BMECs inhibited the proliferative activity of microglial cells, suggesting the therapeutic potential of TLJN on ischemic cerebral vascular disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Brain / blood supply*
  • Cell Hypoxia
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Chemokine CCL4 / metabolism*
  • Culture Media, Conditioned / metabolism
  • Down-Regulation
  • Drugs, Chinese Herbal / administration & dosage
  • Drugs, Chinese Herbal / pharmacology*
  • Endothelial Cells / drug effects*
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology
  • Enzyme-Linked Immunosorbent Assay
  • Glucose / deficiency*
  • Immunohistochemistry
  • Injections
  • Microglia / drug effects
  • Microglia / pathology
  • Microvessels / drug effects*
  • Microvessels / metabolism
  • Microvessels / pathology
  • Neuroprotective Agents / administration & dosage
  • Neuroprotective Agents / pharmacology*
  • Phytotherapy
  • Plants, Medicinal
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, CCR5 / metabolism
  • Time Factors

Substances

  • Chemokine CCL4
  • Culture Media, Conditioned
  • Drugs, Chinese Herbal
  • Neuroprotective Agents
  • Receptors, CCR5
  • tongluo jiunao
  • Glucose