The neuroprotective effects of Tao-Ren-Cheng-Qi Tang against embolic stroke in rats

Background: Combinations of the traditional Chinese and Western medicines have been used to treat numerous diseases throughout the world, and there is a growing body of evidence showing that some of the herbs used in traditional Chinese medicine elicit significant pharmacological effects. The aim of this study was to demonstrate the neuroprotective effects of Tao-Ren-Cheng-Qi Tang (TRCQT) in combination with aspirin following middle cerebral artery occlusion (MCAO)-induced embolic stroke in rats. Methods: A blood clot was embolized into the middle cerebral artery of rats to induce focal ischemic brain injury. After 24h of MCAO occlusion, the rats were arbitrarily separated into five groups and subjected to different oral treatment processes with TRCQT and aspirin for 30days before being evaluated in terms of their neurological behavior using a four-point system. The rats were sacrificed at 30days after drug treatment and the infarct volumes were measured using a 2,3,5-triphenyltetrazolium chloride staining method. Tumor necrosis factor-α (TNF-α), c-Jun N-terminal kinases (JNK), activated caspase-3 and Bax were detected by western blot analysis. The apoptotic cells were identified by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. ROS generation was also measured by electron spin resonance spectrometry. Results: Rats treated with TRCQT alone or in combination with aspirin showed a significantly reduced infarct volume (P<0.001) and improved neurological outcome compared with those treated with distilled water. Rats treated with TRCQT alone (P=0.021) or in combination with aspirin (P=0.02) also showed significantly reduced MCAO-induced expression levels of TNF-α and pJNK (P<0.001) in their ischemic regions. Rats treated with TRCQT alone or in combination with aspirin showed decreased apoptosis by a reduction in the number of TUNEL positive cells, which inhibited the expression of activated caspase-3 (P=0.038) and Bax (P=0.004; P=0.003). TRCQT also led to a significant concentration-dependent reduction in the formation of hydroxyl radicals (P<0.001). Conclusions: TRCQT reduced brain infarct volume and improved neurological outcomes by reducing apoptosis, attenuating the expression of TNF-α and p-JNK, and reducing the formation of hydroxyl radicals in MCAO-induced embolic stroke of rats.