| Summary: | 碩士 === 國立陽明大學 === 傳統醫藥學研究所 === 97 === Local somatothermal stimulation (LSTS) was demonstrated to increase the expression of myocardial heat shock protein 70 (Hsp70) and protected rat hearts against ischemia-reperfusion injury. However, the exact mechanisms of how LSTS works remains unknown. The aim of this study was to test the hypothesis that LSTS at PC 6 induced free radicals formation in the muscle beneath acupoint and subsequently induced Hsp70 expression in the heart.
LSTS was applied 0.5 cm above and onto acupoint Fujie (SP 14) and Neiguan (PC 6), respectively. Under adequate anesthesia, male Sprague–Dawley rat, were treated with LSTS for 1 dose. Following time intervals of 0 min (no treatment),5-, 15-, 30- and 60- min, respectively, reactive oxygen species (ROS) were measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA) fluorometric assay. Nitrate and nitrite (nitric oxide (NO) end products), malondialdehyde (MDA) , cyclic guanosine monophosphate (cGMP) and free radical scavengers were measured by commercial available kits. Western blot analysis for Hsp70 and Nuclear factor erythroid 2–related factor 2 (Nrf2) in the muscle and the heart were also analyzed; the parameters such as serum creatine kinase-MB isoenzyme (CK-MB) and cardiac tropnin I (cTnI) were measured to evaluate the myocardial injury.
The results showed that the generation of ROS, NO end products (nitrate and nitrite) and MDA in 5 min-, the consumption of superoxide dismutase (SOD) in 5 min-, and the consumption of catalase (CAT) and glutathione (GSH) in 15 min- after the application of LSTS suggested that there were free radicals formation in the muscle beneath the acupoint area. The LSTS-induced Hsp70 expression in the muscle were partially blocked by L-NAME treatment. Besides, the formation of ROS, MDA, cGMP and Nrf2 in 5 min- , the increase of serum CK-MB and cTnI in 15 min- and subsequently the generation of SOD in 30 min- suggested that there were free radicals formation in the heart after LSTS treatment on left PC 6. Finally, There were also ROS, NO end products (nitrate and nitrite) and Nrf2 expression in 5 min- after LSTS were noticed in the muscle beneath acupoint PC 6.
We concluded that LSTS induced free radicals formation in the muscle beneath the acupoint PC 6 and, the corresponding organ–heart, which might explain the possible mechanisms of LSTS-induced myocardial protection in rats.
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