ABSTRACT
OBJECTIVE To explore the potential mechanism of the effect of Xuebijing injection （XBJ） on neurological function and survival of rats after cardiac arrest （CA）/cardiopulmonary resuscitation （CPR） based on the S-nitrosoglutathione reductase （GSNOR）/S-nitrosoglutathione （GSNO） pathway. METHODS The CA/CPR rat model was established by ventricular fibrillation. Using a sham operation group as control， high-throughput sequencing was employed to analyze and mine the differentially expressed genes （DEGs）. Enzyme-linked immunosorbent assay was used to determine the contents of GSNOR and GSNO in the hippocampus； the active components of XBJ were screened and subjected to molecular docking analysis with GSNOR. The rats successfully modeled using the same method were divided into model group （n＝30）， inhibitor （GSNOR inhibitor） group （n＝30）， XBJ group （n＝30） and XBJ+inhibitor group （n＝30）， and a sham operation group （n＝30） was set up. Neurological function was evaluated and survival status was recorded at 3 hours， 24 hours and 3 days after the first 89） drug intervention. The contents of GSNOR and GSNO in the hippocampus of rats were determined in each group at the 0191） above time points， and the relationship of the contents of GSNOR and GSNO with modified neurologic severity scale （mNSS） score was analyzed. RESULTS GSNOR coding gene was differentially expressed between the model group and the sham operation group. Compared with the sham operation group， GSNOR content increased significantly in the hippocampus of rats in model group， while GSNO content decreased significantly （P＜0.05）. The active components of XBJ， such as 4- methylenemiltirone and salviolone， could be bound to GSNOR protein， with the binding energy lower than －6 kcal/mol， mainly connected by hydrogen bonds. Animal experiments revealed that mNSS score and GSNOR levels in the hippocampus of rats in the model group were significantly higher than those in the sham operation group （P＜0.05）， while GSNO levels and survival rate were significantly lower than those in the sham operation group （P＜0.05）. The above indexes of rats were improved significantly in administration groups， the mNSS score in the XBJ group was significantly lower than that in the inhibitor group， the content changes of GSNOR and GSNO in the inhibitor group were more obvious than those in the XBJ group， and the various indicators in the XBJ+inhibitor group were significantly better than the XBJ group and the inhibitor group （P＜0.05）. GSNOR content was positively correlated with the mNSS score， and GSNO content was negatively correlated with the mNSS score （P＜0.05）. CONCLUSIONS XBJ can improve the neurological function of rats and enhance their survival rates after CA/CPR， the mechanism of which may be associated with the down-regulation of GSNOR and the up-regulation of GSNO.
ABSTRACT
Objective To explore the neuroprotection of Shuxuetongmai capsule pretreatment, and the effect on the expression of p38 mitogen-activated protein kinase (p38MAPK) in rats with middle cerebral artery occlusion. Methods Ninety-six male SD rats were divided randomly into sham-operated group,ischemia/reperfusion group (I/R),ischemia preconditioning group (IP),and Shuxuetongmai group(n=24). Each group was further randomly divided into 4 subgroups by 3 h, 6 h, 24 h and 72 h after reperfusion, 6 rats in each subgroup. Sham-operated group was only performed artery separation . The middle cerebral artery occlusion (MCAO) model was set up in I/R rats by Longa method. The IP rats were performed for three minutes on the bilateral carotid artery ligation, and formed MCAO model 24 hours later. The rats in the Shuxuetongmai group were pretreated with Shuxuetongmai capsules for 14 days on gavage before the establishment of MCAO model. The neurological deficits were graded in rats by Zea Longa method. Western Blot was used to determine the protein expression of p38MAPK and P-p38MAPK. Tunel method was applied to detect the apoptosis of neurons and the relationship between expression of p38MAPK, P-p38MAPK and apoptosis of neuron. Results No neurological dysfunction appeared in the sham-operated group at each time points, but not for the other groups, which reached the peak at 24 h. Compared with the I/R group, IP group and Shuxuetongmai group presented the mild neurologic function deficiency at different time points in rats (P0.05). The obvious variation of the value of P-p38MAPK/p38MAPK wasn't detected in sham-operated group at different time points, while obviously presented in I/R group, and the ratios of P-p38MAPK/p38MAPK were increased gradually followed with reperfusion, approaching to the highest level at 24 h. Compared with the I/R group, the P-p38MAPK/p38MAPK declined from 3 h and to the lowest level at 24 h of reperfusion, in both IP and Shuxuetongmai groups(P0. 05). Conclusion Shuxuetongmai capsule pretreatment can induce brain ischemic tolerance, attenuate the apoptosis of neurons in cerebral ischemia reperfusion, and improve neurologic function. The mechanism may be related to the inhibition of p38MAPK phosphorylation.
ABSTRACT
This study was aimed to explore the function of c-Jun N-terminal kinase (JNK) signaling pathway in the induction of brain ischemic tolerance, and observe the function of Shu-Xue Tong-Mai (SXTM) capsule pretreatment. Ischemic preconditioning was performed for 3 min on rats to induce cerebral ischemic tolerance. Rat model of cere-bral ischemia reperfusion (the ischemia pretreatment group, I/R group) was established 24 h later. Western blot was used to detect the protein expression of JNK and phosphorylation of c-Jun N-terminal kinase (P-JNK), comparing to the expression with the sham operation group, I/R group and SXTM capsule group. Tunel method was applied to de-tect the apoptosis of neurons. Relationship between expression of JNK, P-JNK and apoptosis of neurons was also studied. The results showed that compared with the model group, expressions of P-JNK in ischemia preconditioning group and SXTM group were declined significantly (P < 0.05); and the apoptosis of neurons quantity was also de-clined (P< 0.05). It was concluded that ischemia preconditioning can decrease the apoptosis of neurons in cerebral ischemia reperfusion, and improve neurologic function. Its mechanism related to the inhibition of JNK signaling path-way. SXTM capsule pretreatment can protect the cerebral by inhibiting the JNK signaling pathway.