Screening of Anti-Hypoxia/Reoxygenation Agents by an in vitro Model. Part 1: Natural Inhibitors for Protein Tyrosine Kinase Activated by Hypoxia/Reoxygenation in Cultured Human Umbilical Vein Endothelial Cells

Abstract

Protein tyrosine kinase (PTK) signaling pathways play important roles in ischemia/reperfusion (I/R) or hypoxia/reoxygenation (H/R) injuries. Inhibition of PTK activation can protect against I/R- or H/R-induced damages. As one part of our work for seeking bioactive compounds from natural sources against I/R or H/R, in the present study we examined the effects of 54 compounds purified from various traditional Chinese herbs on H/R-induced PTK activation by means of an in vitro H/R model in cultured human umbilical vein endothelial cells (HUVEC). The results demonstrated that an increase in PTK activation was induced after 2 h of reoxygenation. Compounds 2 (macrostemososide A), 3 (laxogenin-3-O-β-D-xylopyranosyl-(1 š 4)-α-L-arabinopyranosyl-(1 š 6)-β-D-glucopyranoside), 4 (chinenoside II), 7 (ginsenoside-Rd), 52 (icariin), 53 (icariside), and 54 (icaritin) showed relatively obvious inhibition on this H/R-induced PTK activation. Compounds 5 (β-sitosterol) and 6 (daucosterine), especially 5, completely blocked such an increased activation of PTK induced by H/R. On the contrary, compound 29 (isocumarine) significantly promoted PTK activation further. Moreover, the effects of these compounds on PTK activation were dose-dependent.

Hypoxia/reoxygenation - ischemia/reperfusion - protein tyrosine kinase - tyrosine phosphorylated proteins - bioactive compounds

References

• 1 McMcord  J M.. .  N. Engl. J. Med.. 1985;;  312 159-163
• 2 Gallagher  K P,, Buda  A J,, Pace  D,, Gerren  R A,, Shlafer  M.. .  Circulation. 1986;;  73 1065-1076
• 3 Uraizer  A,, Reimer  K A,, Murry  C E,, Jennings  R B.. .  Circulation. 1987;;  75 1237-1248
• 4 Richard  V J,, Murry  C E,, Jennings  R B,, Reimer  K A.. .  Circulation. 1988;;  78 473-480
• 5 Nejima  J,, Knight  D R,, Fallon  J T,, Uemura  N,, Manders  W T,, Canfield  D R,, Cohen  M V,, Vatner  S F.. .  Circulation. 1989;;  79 143-153
• 6 Patel  B,, Jeroudi  M O,, O'Neill  P G,, Roberts  R,, Bolli  R.. .  Circulation. 1988;;  78 373
• 7 Shirato  C,, Miura  T,, Downey  J M.. .  FASEB J.. 1988;;  2 A918
• 8 Caldwell-Kenkel  J C,, Currin  R T,, Coote  A,, Thurman  R G,, Lemasters  J J.. .  Transpl. Int.. 1995;;  8 77-85
• 9 Terada  L S.. .  Am. J. Physiol.. 1996;;  270 H945-H950
• 10 Chen  Y,, Yu  A,, Saari  J T,, Kang  Y T.. .  Proc. Soc. Exp. Biol. Med.. 1997;;  216 112-116
• 11 Campos-Gonzalez  R,, Kindy  M S.. .  J. Neurochem.. 1992;;  59 1955-1958
• 12 Hayashi  A,, Imai  K,, Kim  H C,, de Juan  E . .  Invest. Ophthalmol. Vis. Sci.. 1997;;  38 372-380
• 13 Hayashi  A,, Koroma  B M,, Imai  K,, de Juan  E . .  Invest. Ophthalmol. Vis. Sci.. 1996;;  37 2146-2156
• 14 Korematsu  K,, Goto  S,, Nagashiro  S,, Inoue  N,, Oyama  T,, Yamada  K,, Ushio  Y.. .  GLIA.. 1995;;  13 147-153
• 15 Korematsu  K,, Goto  S,, Nagashi  S,, Ushio  Y.. .  J. Cereb. Blood Flow Metab.. 1994;;  14 825-830
• 16 Kindy  M S.. .  J. Cereb. Blood Flow Metab.. 1993;;  13 372-377
• 17 Phillis  J W,, Song  D,, O'Regan  M H.. .  Neuroscience Lett.. 1996;;  207 151-154
• 18 Hayashi  A,, Weinberger  A W,, Kim  H C,, de Juan  E . .  Invest. Ophthalmol. Vis. Sci.. 1997;;  38 1193-1202
• 19 Hagar  H,, Ueda  N,, Shan  S V.. .  Kidney Int.. 1997;;  51 1747-1753
• 20 Crawford  L E,, Milliken  E E,, Irani  K,, Zweier  J L,, Becker  L C,, Johanson  T M,, Eissa  N T,, Crystal  R G,, Finkel  T,, Goldschmidt-Clermont  P T.. .  J. Biol. Chem.. 1996;;  271 2683-2687