Sex differences and dynamic changes of oxidative stress markers in stroke patients

Sex differences and dynamic changes of oxidative stress markers in stroke patients

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Katarina Nováková100%100%-
Bc. Karolína Majerová100%100%-
ISBN: 978-80-972360-1-4

Sex differences and dynamic changes of oxidative stress markers in stroke patients

Petra Pazderová1 , Ingrid Žitňanová2
1 Faculty of Mathematics, Physics and Informatics, Comenius University, Bratislava
2 Faculty of Medicine, Comenius University, Bratislava
pazderova.petra@gmail.com

Stroke is the third leading cause of death and the first leading cause of disability in industrially advanced countries [1]. The most frequent stroke type is the ischemic stroke which occurs after a blood clot blocks a brain blood vessel. In the ischemic stroke free radicals can a play crucial role because they can damage important molecules especially lipids. Significant nitrogen and oxygen radicals increase occurs in both, the ischaemic and reperfusion phases of ischemic stroke. If the antioxidant protection of the patient is not strong enough, the patients´ life is seriously endangered. There have not been developed any specific therapies against the acute ischemic stroke (AIS) so far.

In our combined cohort and case control study we have focussed on the evaluation of dynamic changes of oxidative stress markers (protein carbonyls, lipoperoxides) and oxidative stress protector (total antioxidant capacity (TAC)), in blood at three time intervals: 24h after AIS (group A, n = 81), 7-day (group B, n = 57) and 3-month (group C, n = 16) follow-ups. We have also compared plasma samples (group A) obtained from patients with control samples (n = 77), who did not experience any stroke attacks. Comparisons were done in the whole groups as well as between males (n = 45) and females (n = 36). Age influence was also taken into account.

Total antioxidant capacity was measured spectrophotometrically (UV-1800 Shimazu spectrophotometer) by the TEAC (Trolox Equivalent Antioxidant Capacity) assay [2]. Protein carbonyls in plasma were determined by the commercial OxiSelectTM protein carbonyl ELISA kit (Cell Biolabs, USA) according to manufacturer´s instructions. Lipoperoxides in plasma were determined spectrophotometrically (Epoch Microplate spectrophotometer (SN: 130-148) according to El-Saadani assay [3]. Statistical analyses were performed in statistical program StatsDirect 2.7.2.

 We found a significant increase of TAC in AIS patients in (group C) compared to groups B (p < 0.05) and A (p < 0.05). Sex differences in TAC, protein carbonyls and lipoperoxides were not detected, but AIS patients (group A) had marginally significantly higher TAC (p = 0.05) and significantly higher concentrations of lipoperoxides compared to controls (p < 0.05). Females after stroke were significantly older than males (p < 0.001) but their antioxidative status was similar to males after stroke at younger age. In logistic regression model we took age, gender, lipoperoxides, protein carbonyls and TAC into account. Although the strength of the diagnostic test was poor, with sensitivity 71 % and specificity 64 % using cut-off point 0.5, higher oxidative stress and was confirmed.

 Our study confirms the national statistics that the males after stroke are significantly younger than females. Based on our results there are no significant gender differences in protein carbonyls, lipoperoxides and TAC in patients after stroke in spite of age differences. Higher lipid oxidation recorded in AIS patients is the evidence of elevated oxidative stress during the stroke attack. Increased antioxidant capacity in AIS patients 3 months after the attack may be the consequence of better lifestyle (diet rich in antioxidants) or a pharmacological treatment.

Poďakovanie: 

This study was financially supported by the grant agency of the Ministry of health MZ 2012/10-UKBA-10.

Zdroje: 

[1] Premnath, D. et al. Journal of Pharmacy Research, 2012, 5(1), 551-556.
[2] Re, R. et al. Free Radical Biology and Medicine, 1999, 26, 1231-1237.
[3] El-Saadani, M. et al. The Journal of Lipid Research, 1989, 30, 627-630.

Diskusia

Dobrý deň,
Ja mám skôr takú laickú technickú otázku. Zaujímalo by ma ako sa pacienti po NCMP zaraďujú do takejto štúdie, t.j. akým spôsobom sa od nich získava informovaný súhlas a všetky náležitostí pri takýchto akútnych stavoch. A podľa čoho ste vyberali pacientov, aké ste mali inklúzne/exklúzne kritéria.
Ďakujem

Dobrý deň, ďakujem pekne za otázky, informovaný súhlas sa získava od pacientov, keď sa určí typ NCMP – hemorágia alebo ischémia. Ak je schopný pacient komunikovať, získa sa informovaný súhlas od neho, ak nie je schopný komunikovať, získava sa od jeho rodinných príslušníkov. Krv sa im odoberá na klinické vyšetrenie, takže pár ml krvi navyše pacienta nijako nezaťaží.
Pacienti sa zaraďovali do štúdie po absolvovaní klinického vyšetrenia a CT mozgu a po stanovení subtypu mozgovej príhody: hemoragickej alebo ischemickej. Do štúdie boli zaraďovaní aj pacienti s diabetes mellitus, hypertenziou, chronickou renálnou insuficienciou a ischemickou chorobou srdca. Vplyv oxidačného stresu počas náhlej cievnej mozgovej príhody sa u nich plánuje v najbližšej dobe vyhodnotiť. Ako kontroly boli použití dobrovoľníci bez akýchkoľvek akútnych alebo chronických ochorení, ktorí zatiaľ nezažili žiaden ischemický atak.
V našej štúdii sme spolupracovali s I. neurologickou klinikou v Bratislave, odkiaľ sme dostali vzorky krvnej plazmy pacientov a kontrol. Vzhľadom k tomu, že štúdia trvala dva roky a ešte počas merania sa menil počet ľudí zaradených do štúdie, niektoré parametre sme stanovili s nižším/vyšším počtom vzoriek. Pri modeloch logistickej regresie sme potom pracovali len s tými jedincami, ktorým boli zaznamenané všetky sledované parametre. Pri analýzach dynamických zmien markerov oxidačného stresu sme boli obmedzení tým, že mnohí pacienti sa po NCMP na druhý/tretí odber nedostavili, viacerí pacienti NCMP neprežili, dôsledkom čoho bol pokles počtu vzoriek už po týždni, ako aj po troch mesiacoch po NCMP.