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1.
China Pharmacy ; (12): 653-658, 2024.
Article in Chinese | WPRIM | ID: wpr-1013097

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.

2.
Article in Chinese | WPRIM | ID: wpr-404233

ABSTRACT

[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

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