본태성 고혈압 흰쥐의 앞정강근에서 허혈양상화 후 산화질소합성효소와 Akt의 발현
Expressions of Nitric Oxide Synthases and Akt in the Tibialis Anterior Muscle of the Spontaneously Hypertensive Rat after Ischemic Preconditioning
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초록
산화질소는 허혈-재관류 기간에 생성되어 전사인자들을 활성화시킨다고 알려져 있다. 산화질소를 생성하 는데 필요한 산화질소합성효소(nitric oxide synthase, NOS)는 nNOS와 iNOS, eNOS 등 3종류가 있다. Akt는 세포 생존에 중요한 인자로 알려져 있으며, 허혈양상화는 NOS와 Akt의 발현에 영향을 주어 허혈손상을 감소시킨다고 알려져 있다. 또한 NOS의 변화는 고혈압의 발병과 관련이 있으며, 또한 고혈압은 뼈대근육의 피로저항과 수축력 의 감소 같은 변화를 유도한다고 알려져 있다. 본 실험에서 다리에서 실시한 허혈양상화가 본태성 고혈압 흰쥐의 뼈대근육에서 발현되는 NOS 이형과 Akt의 발현에 어떤 영향을 주는지 알아보고자 하였다. 9 주령의 정상 혈압 흰쥐와 본태성 고혈압 흰쥐를 사용하여 대조군과 허혈양상화 3회군, 10회군으로 나누었다. 허혈양상화는 왼온엉덩동맥을 5분 허혈과 5분 재관류를 3회 또는 10회 반복하여 실시하였고, 재관류 시간 0시간 과 0.5시간, 1시간, 3시간, 24시간에 실험동물을 희생시켜 앞정강근을 적출하였다. NOS 이형과 Akt의 발현 변화를 관찰하기 위해, 면역조직화학염색과 Western blot assay를 시행하였고, 다음과 같은 결과를 얻었다. 본태성 고혈압 흰쥐에서 허혈양상화 처치후 nNOS의 발현은 감소, iNOS의 발현은 증가, eNOS의 발현과 Akt 발 현은 감소하였다. 또한 10회의 허혈양상화는 iNOS의 발현을 증가시키고, nNOS와 eNOS, Akt의 발현을 감소시켰 위의 결과를 종합하면 본태성 고혈압 흰쥐에서 반복적인 허혈양상화는 앞정강근에서 iNOS의 발현을 증가시키 며, eNOS와 Akt의 발현을 감소시키는 것을 알 수 있었다. 따라서 고혈압 상태에서 실시하는 반복적인 허혈양상화 는 앞정강근의 허혈 손상을 증가시킬 것으로 생각된다.
Abstract
Nitric oxide (NO) generated during ischemic-reperfusion phase is known to activate various transcriptional factors. NO is generated by 3 nitric oxide synthase (NOS) isoforms, nNOS, iNOS, eNOS. Akt plays a important role of the cell survival. Ischemic preconditioning (IP) reduces ischemic injury by changing of the expression of the NOSs and Akt. The alterations of the NOSs is related to the development of hypertension, and hypertension has been known to induce the skeletal muscle changes including decreased fatigue resistance and contractile force. The purpose of this study was to observe the effect of IP on the expressions of the NOSs and Akt in the skeletal muscle of the spontaneously hypertensive rat (SHR). Nine weeks old male normotensive rat (Wistar-Kyoto rat, WKY) and SHR were divided into control and IP groups. The IP group was further divided into 3 or 10 cycles of IP. For IP, left common iliac artery was occluded 3 or 10 cycles of 5 min of ischemia/5 min of reperfusion using rodent vascular clamp. The animals were sacrificed at 0, 0.5, 1, 3 and 24 hours after reperfusion and the tibialis anterior muscles were removed. The expressions of nNOS, iNOS, eNOS, and Akt were examined with immunohistochemical methods and Western blot analysis. In the SHR, the expression of nNOS was decreased, iNOS was increased and the expressions of eNOS and AKT were decreased, compare to WKY. The expression of iNOS was increased, and nNOS, eNOS and Akt were decreased in SHR by 10IP. In summary, IP increased the expression of iNOS, and decreased the expression of eNOS and Akt in the SHR. It is suggested that hypertension may aggravate injury of the tibialis anterior muscle induced by repetitive ischemic conditioning.
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