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Вивчення впливу гіпоксії та оксиду азоту на кальцієві струми сенсорних нейронів мишей: Автореф. дис… канд. мед. наук / Н.А. Соловйова, НАН України.

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IAOe?IIAEUeIA AEAAeAI?ss IAOE

?INOEOOO O?C?IEIA?? ?i.I.I. AIAIIIEUeOess

___________________________________________________________

Nieiaeiaa Iaoae?y Aiaoie??aia

OAeE 612.81:616.8

AEA*AIIss AIEEAO A?IIEN?? OA IENEAeO ACIOO IA EAEUeOe??A? NO?OIE
NINI?IEO IAE?II?A IEOAE

14.03.03 – ii?iaeueia o?c?ieia?y

Aaoi?aoa?ao aeena?oaoe?? ia caeiaoooy iaoeiaiai nooiaiy eaiaeeaeaoa
iaaee/ieo iaoe

Ee?a- 1999

Aeena?oaoe??th y ?oeiien

?iaioa aeeiiaia o a?aeae?e? caaaeueii? o?c?ieia?? ia?aiai? nenoaie

?inoeoooo o?c?ieia?? ?i. I.I. Aiaiiieueoey

Iaoeiaee ea??aiee: aeaaeai?e IAI Oe?a?ie

Einothe Ieaoii A?eai?iae/

Io?oe?ei? iiiiaioe: aeieoi? iaaee/ieo iaoe, caa?aeoth/ee a?aeae?eii
aenia?eiaioaeueii? ea?ae?ieia?? ?inoeoooo o?c?ieia?? ?i. I. I.
Aiaiiieueoey, aeaaeai?e Iieaaiei Ieaen?e Ieaen?eiae/

aeieoi? a?ieia?/ieo iaoe, canooiiee aee?aeoi?a ?inoeoooo a?io?i?? ?i.
I.I. Iaeeaae?ia Einoe??i Naa??e Ieaen?eiae/

I?ia?aeia onoaiiaa: eaoaae?a ii?iaeueii? o?c?ieia?? Iaoe?iiaeueiiai
iaaee/iiai oi?aa?neoaoo ?i. I.I. Aiaiiieueoey

Caoeno a?aeaoaeaoueny “ 22 ” /a?aiy 1999 ?. i “ 14-00 ” aiaeei? ia
can?aeaii? niaoe?ae?ciaaii? a/aii? ?aaee Ae-96.198.01 i?e ?inoeooo?
o?c?ieia?? ?i. I.I. Aiaiiieueoey IAI Oe?a?ie ca aae?anith:: 252024, i.
Ee?a, aoe. Aiaiiieueoey, 4.

C aeena?oaoe??th iiaeia iciaeiieoenue o a?ae?ioaoe? ?inoeoooeo
o?c?ieia?? ?i. I.I. Aiaiiieueoey, i. Ee?a, aoe. Aiaiiieueoey, 4

Aaoi?aoa?ao ?ic?neaiee “ 21 ” o?aaiy 1999 ?.

A/aiee nae?aoa? niaoe?ae?ciaaii? a/aii? ?aaee

Aeieoi? a?ieia?/ieo iaoe
C.I. Ni?ie?ia-Ia??ia1

CAAAEUeIA OA?AEOA?ENOEEA ?IAIOE

Aeooaeuei?noue i?iaeaie. Aeniea /ooeea?noue ee?oei oeaio?aeueii?
ia?aiai? nenoaie (OeIN) aei a?iien??/?oai?? caaaeueiia?aeiia. ?iiae?
iaa?oue ei?ioei/ani? ci?ie ni?aa?aeiioaiiy i?ae iinoa/aiiyi oa aeo?aoaie
eenith i?eaiaeyoue aei iaiai?ioieo ii?ooaiue ooieoe?e iae?ii?a, ye?
no?aaeaeathoue a iaai?e i??? /a?ac aieea iaaeeeoeo ieneaeo acioo (NO) oa
nii??aeiaieo niieoe, ui ooai?ththoueny, ia ?iii? eaiaee ieaciaoe/ii?
iaia?aie oa aioo??oiueiee?oeiio neaiae?caoe?th. Oae? ii?ooaiiy ooieoe?e
iae?ii?a coiiaeai? oe?eith ieceith yaeu ? i?iaeoeoe?y ieneaeo acioo (NO)
? iaeiei c eiiiiiaio?a o ?o iine?aeiaiino?. ? caaaeueii i?eeiyoei, ui
caaeaaeue iae?ii?a i?e ?oai?? oa a?iien?? a a?eueoino? aeiaaee?a
iia’ycaia c aeoeaaoe??th NMDA ?aoeaioi??a, aeoeaaoe??th eaeueoe??aeo
eaiae?a oa coiiaeaiei oeei ia?aiaaaioaaeaiiyi oeeoiieacie ee?oei ?iiaie
eaeueoe?th. Ianooiia aeoeaaoe?y eaeueoe?e-caeaaeieo neioac ieneaeo acioo
aaaea aei i?aeaeuaiiy aioo??oiueiee?oeiii? eiioeaio?aoe?? NO. Oiio
aaaaea?oueny, ui iaae?aoi?ii oaeiai aiiene/iiai, NMDA –
iiina?aaeeiaaiiai iioeiaeaeaiiy iae?ii?a ? ieneae acioo aai ?iaeoeo eiai
aca?iiae?? c noia?ieneaeai?iiii, ia?ienei?o?eo (ONOO-).

A cia/ii iaioiio nooiai? aea/aii aieea ianoa/? eenith ia iae?iie
ia?eoa?e/ii? nenoaie, cie?aia, ia naini?i? iae?iie aai iae?iie
caaeiuei-ei??ioeaaeo aaiae??a (CEA), ui caaacia/othoue ia?aaea/o
naini?ii? ?ioi?iaoe?? a?ae ?aoeaioi??a aei no?oeoo? OeIN. Oae?
aenia?eiaioaeuei? aeai? ia/eneaii? oa, a e?auiio aeiaaeeo, aeathoue
oaiiiaiieia?/iee iien yaeua aac aeaoae?caoe?? eiai ee?oeiieo iaoai?ci?a.

Iaoa ?iaioe iieyaaea o aea/aii? iiaeeeaino? o/ano? ieneaeo acioo oa
iaeiiai c eiai iaoaaie?o?a – ia?ienei?o?eoo o iaoai?ciao a?iiene/iiai
iioeiaeaiiy iae?ii?a caaeiuei-ei??ioeaaeo aaiae??a ieoae.

Aieiai? caaea/? aeine?aeaeaiiy:

1. Aea/eoe aieea cieaeaiiy ieneaaiaoe?? oa aeieaaee ieenethaaeueiiai
oinoi?eeoaaiiy ia iecueii- (IIE) oa aenieiii?iaia? (AIE) eaeueoe??a?
eaiaee a iae?iiao caaeiuei-ei??ioeaaeo aaiae??a ieoae.

2. Aea/eoe aieea aooaioe/iiai ieneaeo acioo, eiai aeiii??a oa
ia?ienei?o?eoo iecueei- oa aenieiii?iaia? eaeueoe??a? eaiaee a iae?iiao
CEA ieoae.

Iaoeiaa iiaecia iaea?aeaieo ?acoeueoao?a. Aia?oa aea/aia ae?y
aooioe/iiai ieneaeo acioo oa eiai iio?aeiiai – ia?ienei?o?eoo – ia
iecueeiii?iaia? oa aenieiii?iaia? eaeueoe??a? eaiaee a iae?iiao CEA
ieoae. Aea/aii aieea ??cieo aeiii??a ieneaeo acioo ia IIE oa AIE a
iae?iiao CEA ieoae. Anoaiiaeaii, ui oe? aeaa oeie eaiae?a a a?eueoino?
iae?ii?a na?aaeiueiai ae?aiao?o CEA ieoae iathoue aenieo ?acenoaioi?noue
aei a?iien?? oa, aieaeii/an, an? aiie ? aeniei/ooeeaeie aei aooaioe/iiai
NO. I?iaeaiiino?iaaii, ui ii?yae c iae?iiaie, eaeueoe??a? no?oie yeeo ?
?acenoaioieie a?iien??, CEA ieoae i?noyoue oaeiae iacia/io e?euee?noue
iae?ii?a, eaeueoe??a? eaiaee yeeo /ooeea? aei cieaeaiiy nooiaith
ieneaaiaoe?? ciai?oiueiai ?ic/eio. A oaeeo iae?iiao ianoa/a eenith
i?eaiaeeoue aei iai?ioiiai ?ia?aoaaiiy iecueei- oa aenieiii?iaiaeo
eaeueoe??aeo no?oi?a. Aia?oa iieacaii, ui ia?ienei?o?eo iaiai?ioii
aeieo? iaeaeaa oeie aeuacaaaeaieo no?oi?a.

Oai?aoe/ia oa i?aeoe/ia cia/aiiy iaea?aeaieo ?acoeueoao?a. I?iaaaeai?
aeine?aeaeaiiy nooo?ai aeiiiaiththoue no/ani? oyaeaiiy i?i iaoai?cie
o/ano? NO oa iaeiiai c eiai iio?aeieo – ia?ienei?o?eoo – a ?aaoeyoe??
eaeueoe??aiaii aiiainoaco a Iiaeaeueo? aeine?aeaeaiiy /ooeeaino? IIE oa
AIE aei oa?iaeieia?/ieo ?a/iaei a oiiaao cieaeaii? ieneaaiaoe?? iiaeooue
i?eaanoe aei aeyaeaiiy e?ea?nueeeo cania?a, ui aieeaathoue ia aeaeeaaiiy
iae?ii?a i?e ?o ?oai?/iiio iioeiaeaeaii?.

Iniaenoee aianie caeiaoaa/a. I?eaiooaaiiy oa?eaioaoeaii ?cieueiaaieo
iae?ii?a CEA ieoae oa an? aeaeoi?o?c?ieia?/i? aenia?eiaioe ii
aeine?aeaeaiith aieeao ieneaeo acioo oa a?iien?? ia eaeueoe??a? no?oie
aoee aeeiiai? aaoi?ii iniaenoi. Caeiaoaa/ a?aa aeoeaio o/anoue o
iaaiai?aii? ?acoeueoao?a oa oi?ioethaaii? aeniiae?a.

Ai?iaaoe?y ?acoeueoao?a aeena?oaoe??. Iniiai? iieiaeaiiy ?iaioe
aeeeaaeaeenue oa iaaiai?thaaeenue ia nai?ia?ao ?inoeoooo o?c?ieia?? ?i.
I.I, Aiaiiieueoey IAI Oe?a?ie (1996-!998 ??.), XV c‘?cae? Oe?a?inueeiai
o?c?ieia?/iiai oiaa?enoaa (Aeiiaoeuee, 1998), eiioa?aioe??
o?c?ieia?/iiai oiaa?enoaa Aaeeeia?eoai?? (E?aa?ioeue, 1998).

Ioae?eaoe??. Ca ?acoeueoaie ?iaioe iioae?eiaaii o?e noaoo? oa oace
aeaio aeiiia?aeae.

No?oeoo?a oa ianya aeena?oaoe??. Aeena?oaoe?y neeaaea?oueny c? anooio,
iaeyaeo e?oa?aoo?e, iaoa??ae?a oa iaoiae?a aeine?aeaeaiiy, ?acoeueoao?a
aeine?aeaeaiue, iaaiai?aiiy ?acoeueoao?a, aeniiae?a oa anooio
aeei?enoaieo aeaea?ae c? 162 iaeiaioaaiue. ?iaioa aeeeaaeaia ia 105
noi??ieao oa ?ethno?iaaia 11 ?enoieaie.

INIIAIEE CI?NO AeENA?OAOe??

Iaoa??aee oa iaoiaee aeine?aeaeaiue.

A aenia?eiaioao aeei?enoiaoaaeenue iae?i?ei? ieo? a?eii 4 oeaei?, ui
oo?eioaaeenue ia noaiaea?oi?e eaai?aoi?i?e ae??o?.

Iaoiae oa?iaioaoeaiiai aeae?eaiiy iae?ii?a CEA. I?ney ao??iiai ia?eico
oa aeaeai?oaoe?? oaa?ei aeae?eyee a?oaei? oa e?eaeia?
caaeiuei-ei??ioeaa? aaiae?? a /aoeo Iao?? c ioieiaeaeaiei aei +4N
iiaeeo?eiaaiei ?ic/eiii Oe?iaea oaeiai neeaaeo (a iI): NaCl – 140, KCl –
4, MgCl2 – 2, CaCl2 – 2, HEPES – 10, aetheica – 10 (?I = 7.4). I?ney
oeueiai aaiae?? ia?aiineee o oeaeii c 2 ie i?ae?a??oiai aei 36N ?ic/eio
oaeiai ae neeaaeo ?c aeiaeaaaiiyi 1 ie/ia i?ioaace (oeiXIV, Sigma, NOA)
oa 1 ia/ie eieaaaiace (Wortington, NOA) oa aeo?eioaaee a oa?iinoao? i?e
aeacai?e oaiia?aoo?? ia i?ioyc? 11 oaeeei, i?ney /iai 10 oaeeei
a?aeieaaee a aacoa?iaioiiio ?ic/ei?. Aeey io?eiaiiy ee?oeiii? noniaic??
aaiae?? aeenioe?thaaee ca aeiiiiiaith iieaaeaieo Ianoa??anueeeo i?iaoie.
Io?eiaio noniaic?th ia?aiineee a aenia?eiaioaeueio eaia?o oa aeo?eioaaee
a oa?iinoao? i?e 36N ia i?ioyc? 1 aiaeeie aeey i?ee??ieaiiy ee?oei aei
aeia eaia?e.

Aeaeo?io?c?ieia?/i? aei??thaaiiy.

?a?no?aoe?th eaeueoe??aeo no?oi?a iaia?aie ?cieueaaieo iae?ii?a
i?iaiaeeee iaoiaeii o?enaoe?? iioaioe?aeo (“iao/-eeaii”) a eiio?ao?aoe??
“oe?e? ee?oeia”, aeei?enoiaoth/e neeyi? i?e?ii?iaoee, ui aeaioiaeyeenue
o aea? noaae?? ca iienaiith iaoiaeeeith (Neher & Sakmann, 1976; Neher et
al., 1978; Hamill et al., 1981). CsCl – 130, TEACl – 20, MgCl2 – 5, ATP
– 5, EGTA – 2, BAPTA – 2, HEPES – 10 (?I – 7.3). Aeey aeeeth/aiiy
iao???aeo no?oi?a ?iie iao??th cai?uoaaee ?iiaie oie?ia oei??aeo. I?ney
caiiaiaiiy i?e?ii?iaoee aioo??oiueiee?oeiiei ?ic/eiii ?? ii?? neeaaeaa
2-3 IIi. Aeey naia?aoe?? IIN oa AIN aoee aeei?enoai? a?aeiia?aei?
aenia?eiaioaeuei? i?ioieiee. IIN, ui oaeaeei ?iaeoeao?oueny, io?eioaaee
oeyoii aeaiiey?ecaoe?? iaia?aiee ee?oeie aei –35 iA a?ae i?aeo?eioth/iai
iioaioe?aeo –80 iA, a AIN – oeyoii aeaiiey?ecaoe?? aei 0 iA a?ae
i?aeo?eioth/iai iioaioe?aeo –50 iA. *a?ac eaia?o iino?eii i?ioiaeea
ciai?oiueiee?oeiiee ?ic/eic? oaeaee?noth 0.5 – 1 ie/oa oa c
oaiia?aoo?ith 36N. Iine?aeiaiee ii?? eiiiainoaaany aia?aoii ia 50-70%.
Aeo?aeiee neaiae i?aeneethaa/a o?eueo?oaaee ia /anoio? c??co 2 eAoe ca
aeiiiiiaith 4-iiethniiai o?eueo?a Aanaey. Ca?? aeaieo oa ?o aiae?c
aeeiioaaeenue ca aeiiiiiaith iaeao?a i?ia?ai ?Nlamp, aa?n?y 6.0.3 (Axon
Instruments Inc., NOA), Origin, aa?n?y 4.0 (MicroCal software, NOA).

Iaoiaee aei??thaaiiy iai?oae eenith a ?ic/eiao. Aeey aei??thaaiiy ?I2 a
aenia?eiaioaeuei?e eaia?? oa aooa?ieo ?ic/eiao aeei?enoiaoaaeenue
noaiaea?oi? iiey?ia?ao?/i? iaoiaee. Aei ?aai/i? eaia?e aoa i?eeaooiaaiee
a?aee?eoee noa?e/iee ieaoeiiaee aeaeo?iae ae?aiao?ii 100 iei oa
oei?n??aiee ?iaeeoa?aioiee aeaeo?iae. sse iiey?ia?ao, aeei?enoiaoaaany
i?eeaae LP-7A (*aoineiaae?y). Oaeiae aei??thaaiiy ?I2 a ?ic/eiao
aeeiioaaeinue ca aeiiiiiaith aaciaiai aiae?caoi?a AAN-1 (aeai?y) c
aeei?enoaiiyi cae?eoiai i?eaoeiiaiai aeaeo?iaea oeia Eea?ea.

Iaoiaee aei??thaaiiy eiioeaio?aoe?? ieneaeo acioo a ?ic/eiao. Aei??
eiioeaio?aoe?? NI a aooa?ieo ?ic/eiao i?iaiaeeany ca aeiiiiiaith
ISO-NO-iao?a o??ie World Precision Instruments Inc. (NOA) c iaia?aiiei
NO-naini?ii ae?aiao?ii 2 ii. I?eioeei aei??o eiioeaio?aoe?? ieneaeo
acioo ciniiaaiee ia aiia?iiao?e/iiio iaoiae?: NO oeyoii aeeooc??
i?ioiaeeoue /a?ac naeaeoeaio iaia?aio aeaeo?iaea-naini?a, aea
ieeneth?oueny ia ?iai/iio aeaeo?iae?, ui ? i?eaiaeeoue aei iiyae
aeaeo?e/iiai no?oio, aiie?ooaea yeiai i?iii?oe?iiaeueia eiioeaio?aoe??
NO a ?ic/ei?.

Iaoiae i?eaiooaaiiy aooaioe/iiai ieneaeo acioo. A aenia?eiaioao aoa
aeei?enoaiee aiaeiee ?ic/ei aaciiiae?aiiai NO (oaeee NO ?iaiothoue
aooaioe/iei ia a?aei?io a?ae oiai, ui iaea?aeo?oueny c ?a/iaei –
aeiii??a). Aeey i?eaiooaaiiy aiaeiiai ?ic/eio NO /a?ac aa?iaoe/iee
oeaeii (20 ie) i?ioyaii 30 oa i?iaeoaaee 99% NO. Iio?i oeae oeaeii
caiiaithaaee 20 ie aea?ii?ciaaii? oa aeaieneaaiiaaii? aiaee, i?ney /iai
5 ie aaciiiae?aiiai NO ca aeiiiiiaith oi?eoey ?i’?eooaaee a oeae ia’?i
aiaee, io?eioth/e oaeei /eiii ?ic/ei NO, eiioeaio?aoe?y yeiai noaiiaeea
aeecueei 3 iI.

Neioac ia?ienei?o?eo?a. Aeey i?eaiooaaiiy aaciaiai ?ic/eio
ia?ienei?o?eoo aeei?enoiaoaaeenue ianooii? ?ic/eie: 1) ?ic/ei A, 0.6 I
NaNO3; 2) ?ic/ei A, 0.72 I I2I2 + 0.68 I HCl; 3) ?ic/ei N, 1.25 I NaOH.
I?ney ci?ooaaiiy i?aenoieiaee ?ic/ei ia?iaeyany ii?ioeii ieeno iaai?y oa
o?eueo?oaaany. Eiioeaio?aoe?y ia?ienei?o?eoo a na?aaeiueiio
aei??aithaaeany 140 20 iI. Aaciaee ?ic/ei ?icaiaeeany aaciina?aaeiuei
ia?aae aeiaeaaaiiyi a ?iai/o eaia?o.

?acoeueoaoe aeine?aeaeaiue.

?iciiae?e iae?ii?a caaeiuei-ei??ioeaaeo aaiae??a ca ?ici??aie.

Ia ?en.1 cia?aaeaia a?noia?aia ?iciiae?eo ca ?ici??aie iae?ii?a
oa?iaioaoeaii ?cieueiaaieo iae?ii?a CEA. Ai?ieneiaoe?y aeaii? a?noia?aie
ooieoe??th Aaona (ioieoe?ia e?i?y ia ?enoieo) aeicaieeea aeyaeoe o?e
a?oie ee?oei c na?aaei?ie ?ici??aie 29, 40 oa 51 iei, ui aeia?a
ocaiaeaeo?oueny c aeaieie ?ioeo aaoi??a (Scroggs & Fox, 1992). Aeey
aenia?eiaio?a a?aeae?aeeny ee?oeie, ?ici??e yeeo aoee aeecuee? aei
aeuacaaaeaii? na?aaeiuei? aaee/eie – 40 iei. ?o oiiaii iaceaaee
na?aaei?ie. Cia/aiiy iioaioe?aeo niieith oaeeo iae?ii?a ciaoiaeeeenue o
iaaeao a?ae –75 iA aei –50 iA. A ee?oeiao oe??? a?oie ?a?no?oaaeenue ye
iecueeiii?iaia? (IIN) oae ? aenieiii?iaia? (AIN) eaeueoe??a? no?oie
(INa). Ia ?en. 2 a?aeia?aaeaia caeaaei?noue iai?oae eenith a
ciai?oiueiio ?ic/ei? a?ae o?eaaeino? eiai i?iaeoaaiiy aaciiiae?aiei
acioii. Aeaeii, ui i?ney 3-4 oaeeei ?ioaineaiiai i?iaeoaaiiy ??aaiue ?I2
cieaeo?oueny c? 145-147 ii ?o.no. aei i?i?iaeueieo cia/aiue ii?yaeeo 5 –
7 ii ?o.no. oa i?aeo?eio?oueny ia oaeiio ??ai? ia i?ioyc? anueiai /ano
i?iioneaiiy aaco. Ia?ooc?y aenia?eiaioaeueii? eaia?e ?ic/eiii c ?I2 5-7
ii ?o.no. aeicaiey? iaoe o ?? aeia cia/aiiy ?I2 ??ai? 10 – 12 ii ?o.no.,
a i?e ?I2 a ia?ooc?ei?e iinoaeei? 10 ii ?o.no., ?I2 a ?iai/?e eaia??
aei??aithaaei 20 – 22 ii ?o.no. A?aeiii, ui e?eoe/ia aaee/eia aeey
iiceiaiai ?I2 aei??aith? 19 – 23 ii ?o.no. (Doppenberg et al. 1998),
oiio naia oae? oa iaio? aaee/eie e aeei?enoiaoaaeenue iaie o a?eueoino?
aenia?eiaio?a i?e iiaeaethaaii? a?iiene/ieo oiia. O 80% aeine?aeaeaieo
iae?ii?a cieaeaiiy nooiaith ieneaaiaoeee ciai?oiueiai ?ic/eio c? 145 aei
20 ii ?o.no. ia aeeeeeaei i?yeeo ci?i eaeueoe??aeo no?oi?a, a cieaeaiiy
ua ia 10 ii ?o.no. i?eaiaeeei aei iacia/iiai ciaioaiiy aiie?ooaee ye
IIN, oae ? AIN, yea, iaeiae, ia aoei noaoenoe/ii a??ia?aeiei (?en. 4 A,
A). ?ieoaaoe?y ?ioeo 20% iae?ii?a a ?ic/ei? c ?I2 20 ii ?o.no. aeeeeeaea
iai?ioia cieaeaiiy iaeneiaeueii? aiie?ooaee eaeueoe??aiai no?oio /a?ac
IIE a na?aaeiueiio c 3.1 0.2 aei 2.3 0.2 iA (n=6, ?

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