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目的用硝普钠(NO供体)干预鸡胚,通过检测二氧化硫/天冬氨酸氨基转移酶(SO2/GOT)体系的变化,探讨内源性NO对早产仔鸡动脉导管中SO2/GOT体系的作用。方法随机数字表法将恒温(37.8℃)、恒湿(55%)孵化胚龄为13 d的清洁级新鲜无破损白色来亨鸡种蛋60枚随机分成4组(每组15枚):空白对照组和3个硝普钠处理组(小剂量组,中剂量组,大剂量组)。空白对照组单纯用空针穿刺种蛋尿囊腔,小、中、大剂量组待胚龄满15 d时分别用1μg/g、2.5μg/g、5μg/g的硝普钠注射种蛋尿囊腔,注射后蜡封种蛋气室端穿刺孔后继续孵化。待胚龄满18 d时将种蛋破壳取出仔鸡断头法处死后,立即解剖,仔鸡心脏采血1 ml后立即取仔鸡动脉导管组织。用去蛋白法测定仔鸡血浆SO2含量,用实时荧光定量PCR法测定动脉导管组织中GOT mRNA的表达,用Westernblot法测定仔鸡动脉导管组织中GOT蛋白的表达情况。结果血浆SO2含量对照组,小、中、大剂量组分别为(33.68±6.57)mol/L、(40.02±6.09)mol/L、(48.71±7.12)mol/L、(59.63±8.07)mol/L;动脉导管组织GOT mRNA表达量的2-△△Ct值对照组,小、中、大剂量组分别为1.02±0.11、1.39±0.13、4.72±0.20、7.63±0.13。早产仔鸡血浆SO2含量、动脉导管GOT蛋白和GOT mRNA表达量均随硝普钠注射剂量的递增而递升,差异均有统计学意义(P均<0.05)。结论 GOT蛋白和GOT mRNA在仔鸡动脉导管组织中均有表达,硝普钠对早产仔鸡动脉导管中SO2/GOT体系在一定剂量范围内有浓度依赖性的促进作用。
Objective To investigate the effect of endogenous nitric oxide (NO) on SO2 / GOT system in arterial duct of premature chickens by intervention of sodium nitroprusside (NO donor) effect. Methods A random number table was used to divide 60 fresh clean white Leghorn chickens of embryonic age 13 days (37.8 ℃) and constant humidity (55%) into 4 groups (15 in each group): blank control Group and three sodium nitroprusside groups (low dose group, middle dose group, high dose group). The blank control group was punctured with empty needles only. The small, medium and large dose groups were treated with sodium nitroprusside (1μg / g, 2.5μg / g, 5μg / g) After the injection, the wax seal eggs continue to hatch after puncture. To be embryo age at 18 days when the eggs will be broken out of hatchlings chicken decapitation after sacrifice, immediately dissected, 1ml of blood collected immediately after the heart of chickens arteries of the ductus arteriosus. The content of SO2 in plasma of chicken was determined by deproteinization method. The expression of GOT mRNA in ductus arteriosus was determined by real-time fluorescence quantitative PCR. The GOT protein expression in ductus arteriosus was determined by Western blot. Results Plasma levels of SO2 were (33.68 ± 6.57) mol / L, (40.02 ± 6.09) mol / L, 48.71 ± 7.12 mol / L, 59.63 ± 8.07 mol / L; the expression of GOT mRNA in the ductus arteriosus was 1.02 ± 0.11, 1.39 ± 0.13, 4.72 ± 0.20 and 7.63 ± 0.13 in the control group, small, medium and large dose groups respectively. Plasma SO2 content, GOT protein and GOT mRNA expression in preterm litter chickens increased with increasing doses of sodium nitroprusside, the differences were statistically significant (P all <0.05). Conclusion Both GOT protein and GOT mRNA are expressed in ductus arteriosus of chickens. Sodium nitroprusside promotes the concentration-dependent manner of SO2 / GOT in arterial catheter of premature chickens in a dose range.