Hideaki Higashino, Atsuko Niwa, Masaki Tabuchi, Kana Ooshima, and Hiroshi Sakaue Department of Pharmacology, Kinki University School of Medicine, Osaka-Sayama, 589-8511, Japan.Long-term Voluntary Exercise Decreased the Incidence of Apoplexy and Elongated the Lifespan through Activation of eNOS and Inhibition of Inflammatory Signaling Pathway in Stroke-Prone SHR

Intervention Therapy for Hypertension Guidance for Improvement of Life Style a. Food (low salt, low calorie, much fiber) b. Physical exercise c. Save the body weight d. To avoid much stress e. Enjoy the daily lifeDrug therapy a. Early treatment b. Select the appropriate drugs for prevention of AS

Objective:

Clinical evidences show that exercise exerts atheroprotective or beneficial effects on cardiovascular events. Precisely causative mechanisms, however, are still unknown. Therefore, the hypothesis that endurance voluntary exercise decreases the inflammatory signaling through eNOS induction and ROS inhibition was assessed in SHRSP.

Methods

Animals:Male SHRSP aged 6-week-old at pre-hypertensive stage Groups: 1. Voluntary wheel-running (WR): 2 to 3 km running/day 2. sedentary control (SED): in the cage without runningDuration:8 weeks

Analyses: Thoracic Aortae: NOS expression, eNOS activity, oxidative stress Akt, eNOS, phosphorylated ones by western blotting NADPH oxidase mRNA by RT-PCR. Activities of eNOS by using [3H]l-arginine Blood: Superoxide (O2-) production by flow cytometer using DHEPlasma: sICAM-1, MCP-1, 8-iso-PGF2by ELISA

Observation of the occurrence of apoplexy: keeping them until the death

140.0160.0180.0200.0220.0240.0260.0280.0Age (weeks)Systolic blood pressure 678910111213141516(mmHg)** P

050100150200050100Age (Days)(%)SEDENTARY (n=15)EXERCISE (n=13) EX vs. SED; P=0.016Stroke morbidityStart of exercise Incidents of Stroke determined by stroke scores in SED and WR SHRSP

0.51100200Age (days)Periods of Life-Span in Sedentary and Exercised SHRSPSurvivingp

SEDENTARY EXERCISE

00.10.20.30.4SEDWR051015202530SEDWR(mm)(%)Thickness of media Collagen area of vessel wall **p

Expression of angiotensin (AT)1 receptors and AT2 receptors in the aortas of SED and WR SHRSPSEDENTARY AT1REXERCISE AT1RSEDENTARY AT2REXERCISE AT2R

00.511.52SEDWRAT1R / AT2Rn=66*p

Nox1Relative intensity (arb. units)mRNA/ 18S rRNASEDWR 44mRNAproteinSEDWR58P

00.20.40.60.811.21.41.61.8Relative intensity (arb. units)p-Akt/AktSEDWRp

ROS production05001000150020002500SEDWR55

P

0246810121416[3H]l-citrulline (nmol/mg/min)0100200300400500600cGMP (fmol/mg protein)NOS activitySEDWRcGMPComparison of NOS activities and cGMP production in the AortaeSEDWRp

00.20.40.60.811.21.4Nitrotyrosine (arb. units)020406080100120140Dihydroethidium (MFI)Nitrotyrosine in AortaeSEDWRMFI in the BloodSEDWRNitrotyrosine contents in Aortae and MFI by DHE in the Bloodp

00.20.40.60.811.21.41.6SEDWR00.20.40.60.811.21.41.61.8p-Akt/Aktp

0123456SEDWRTGF-(ng/ml)n=64(Mean SEM)Changes of serum TGF- levels after exercise

0100200300400500600SEDWRhigh sensitive CRP(mg/ml)p

(ng/ml)(ng/ml)0246810SEDWRMCP-187p

Physical Exercise PI3KAkteNOSNOcGMPPrevention of Cardio-vasculitisAng II, TNF- PLC, PLDNAD(P)HoxidaseROSMAP kinasesONOO-MCP-1ICAM-1MAT1 Rs/AT2 RsVascular dilatationHypertensionApoplexyShear stressFibrosis

Conclusions: Data showed that exercise could protect oxidative stress-induced cell injury or inflammation by an interaction with signaling molecules such as ASK1 /JNK/ p38MAPK through NO production and inhibition of superoxide production.

Then, voluntary exercise significantly attenuated the changes of vascular remodeling, delayed stroke events and elongated the lifespan in exercised rats.