Expression of signaling molecules associated with apoptosis in human ischemic stroke tissue.
There is growing evidence that, because of the highly significant differences in gene activation/protein expression between animal models of stroke and stroke patients, the current treatment strategies based on animal stroke models have been unsuccessful. Therefore, it is imperative that the pathobiology of human stroke be studied. As a first step here, Western blotting and immunohistochemistry were employed to examine expression and tissue localization of key apoptotic proteins in infarct and peri-infarcted (penumbra) from grey and white matter in human postmortem tissue of 18 patients who died between 2 and 37 d after stroke caused by large vessel disease. The contralateral hemisphere was used as a control. JNK1, JNK2, and p53 were upregulated in the majority of samples, whereas Bcl-2, caspase-3, active caspase-3, phosphorylated p53 (p-p53), phosphorylated JNK1 (p-JNK1), and phosphorylated JNK2 (p-JNK2) were upregulated in approximately half of the samples. JNK1 expression was positively correlated with JNK2 expression in grey and white matter infarct and penumbra, whereas active caspase-3 levels were positively correlated with p-JNK2 levels in grey and white matter infarct. Using indirect immunoperoxidase staining of paraffin-embedded sections, active caspase-3 was found in infarcted neurons that co-localized with TUNEL-positive cells. p-JNK localization in the nuclei of TUNELpositive cells with the morphological appearance of neurons from infarct and penumbra was also demonstrated. The use of Kaplan Meier survival data demonstrated that the presence of Bcl-2 in penumbra of grey matter correlated significantly with shorter survival (p = 0.006). In conclusion, the present study has identified significantly altered expression of apoptotic proteins in human stroke tissue and shown that the presence of Bcl-2 in penumbra of grey matter has prognostic value. It is tempting to suggest that further studies of apoptotic proteins in human stroke may lead to identification of novel targets for drug discovery.
Mitsios
Nicholas
N
Department of Biological Sciences, Manchester Metropolitan University, Chester St, Manchester M1 5GD, UK.
Gaffney
John
J
Krupinski
Jerzy
J
Mathias
Richard
R
Wang
Qiuyu
Q
Hayward
Stuart
S
Rubio
Francisco
F
Kumar
Patricia
P
Kumar
Shant
S
Slevin
Mark
M
D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis; D001921:Brain / Q000473:pathology; D002545:Brain Ischemia / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D008875:Middle Aged; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D015398:Signal Transduction; D020521:Stroke / Q000473:pathology*; D015533:Transcriptional Activation
D019253:Proto-Oncogene Proteins c-bcl-2; D053148:Caspase 3