bnormalities of the function of the autonomic nervous system are frequent and often serious complications in patients with diabetic mellitus (DM), 1 and may be exacerbated by cardiovascular dysfunction. Power spectral analysis of heart rate (HR) variability (HRV) has been shown to be a quantitative and non-invasive method for assessing the effects of the sympathetic and parasympathetic components of autonomic nervous activity of the cardiovascular system. 2 Power spectral analysis of HRV is also a powerful tool for determinating the autonomic nerve tone in patients with diabetic neuropathy, and fast Fourier transform is statistically more robust. 3 However, time domain measurement of the RR intervals is a possible weakness and bottleneck, 4 so we decided to measure frequency domain (power spectral analysis) alternatively with the time domain of RR intervals in order to investigate autonomic nervous function in rats. A telemetric monitoring system for ECG has been developed for small animals and, moreover, sophisticated software permits analysis of HRV for long-term ECG recording even for species with a rapid heart beat. Therefore, analysis of HRV in rats can provide specific information concerning the autonomic nervous activity underlying control of the cardiovascular system in diabetic conditions without any effects of anesthetic agents or changes in environment.
Methods
Animals and HousingThe study animals were 6 spontaneously diabetic male
Circulation Journal Vol.66, March 2002Kob rats, 15-19 months old and weighing 300-320 g, and 9 male Wistar rats of the same age. All rats were purchased from Seac Yoshitomi Limited Ltd (Tokyo, Japan) and were cared for humanely in accordance with the Principles of Laboratory Animal Care formulated by the National Society for Medical Research and the Guide for the Care and Use of Laboratory Animals published by the National Institute of Health (NIH publication No. 85-23, revised 1996). The rats were housed in individual cages with wood shavings in a light-proof room (TB181 or TB182; National, Osaka, Japan) under a light -dark cycle (LD 12:12; lights on at 08.00 h) that was maintained under constant temperature (24°C). The animals had free access to water and a standard diet. The diabetic state of the Kob rats was confirmed by the cardinal symptoms of loss of body weight, polydipsia, polyuria, polyphagia, cataract, increased fasting blood sugar (FBS) and ruffled hair.
Surgical ProcedureAnimals were anesthetized with 50 mg/kg of intraperitoneal pentobarbital and a telemetric ECG radio-transmitter (25×15×9 mm in size and 7 g in weight; TA11CTA-F40, Data Sciences International, St. Paul, MN, USA), was placed in a dorsal subcutaneous pouch. The paired wire electrodes for the pericardial bipolar leads (apex-base leads) were placed under the skin of the dorsal and ventral thorax and the skin was sutured. ECG records were initiated 3 days after surgery to avoid any possible effects of residual anesthetic or the surgery.
ECG Recordings and Data AnalysesEach rat cage was placed on top...