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目的探讨磷酸鸟苷环化水解酶1(GCH1)基因对房颤犬心房自主神经重构的作用。方法健康成年杂种犬24只随机分为右心房快速起搏组(A-TP组)、对照组、GCH1过表达组和GCH1沉默组,每组6只。A-TP组以400次/min持续右心房起搏4周。GCH1过表达组在右上脂肪垫(RSFP)内注射含GCH1过表达的慢病毒。GCH1沉默组在RSFP内注射含GCH1沉默的慢病毒。感染2周后处死动物,剪取RSFP周围小于0.5 cm的心房组织。分别采用qRT-PCR法检测mRNA的表达,Western blotting法检测蛋白的表达,免疫组化法检测神经密度以及Elisa法检测四氢生物喋呤(BH4)含量。结果 4周后,A-TP组较对照组GCH1表达明显下降(P<0.05)。与对照组相比,GCH1过表达组心房有效不应期明显延长,RSFP中PGP9.5表达下降明显,BH4含量增加。与对照组相比,GCH1沉默组RSFP中PGP9.5表达明显上升,BH4含量减少。结论右房快速起搏导致GCH1基因表达下调,GCH1可能通过增加BH4含量抑制房颤自主神经重构。
Objective To investigate the effect of guanosine monophosphate hydrolase 1 (GCH1) gene on the remodeling of atrial autonomic nerve in atrial fibrillation dogs. Methods Twenty-four healthy adult dogs were randomly divided into A-TP group, control group, GCH1 overexpression group and GCH1 silencing group, with 6 rats in each group. The A-TP group continued right atrial pacing for 4 weeks at 400 beats / min. The GCH1 overexpression group was injected with a lentivirus containing GCH1 overexpression in the upper right fat pad (RSFP). The GCH1 silencing group was injected with a lentivirus containing GCH1 silencing in the RSFP. The animals were sacrificed 2 weeks after infection and the atrial tissue smaller than 0.5 cm around the RSFP was cut off. The expression of mRNA was detected by qRT-PCR, the protein expression by Western blotting, the nerve density by immunohistochemistry and the content of tetrahydrobiopterin (BH4) by Elisa method. Results After 4 weeks, the expression of GCH1 in A-TP group was significantly lower than that in control group (P <0.05). Compared with the control group, GCH1 overexpression group atrial effective refractory period was significantly prolonged, PFP9.5 expression decreased significantly in RSFP, BH4 content increased. Compared with the control group, the PGP9.5 expression in RSFP of GCH1 silencing group increased significantly and the content of BH4 decreased. Conclusions Right atrial pacing leads to down-regulation of GCH1 gene expression. GCH1 may inhibit autonomic remodeling of atrial fibrillation by increasing BH4 content.