L-精氨酸对宫内发育迟缓胎鼠胰岛素样生长因子及其结合蛋白表达的影响

Effects of L-arginine on the expression of insulin-like growth factors and insulin-like growth factor binding protein 3 in rats with intrauterine growth retardation

摘要:

目的：宫内发育迟缓(IUGR)儿常有脑发育的异常,L精氨酸具有舒张血管、增加胎盘血流的作用,可用于改善胎盘缺氧状态,促进胎儿生长发育。用被动吸烟法制作孕鼠IUGR模型,孕8～20d给予不同剂量L精氨酸,了解其对宫内发育迟缓胎鼠脑内胰岛素样生长因子及其结合蛋白表达的影响,并探讨L精氨酸的作用机制。方法：孕鼠随机分为4组:对照组、模型组、L精氨酸小剂量和大剂量防治组,每组9只。孕21d剖宫取胎,应用酶联免疫吸附法检测各组胎鼠脑组织胰岛素样生长因子Ⅰ(IGFⅠ)、胰岛素样生长因子Ⅱ(IGFⅡ)、胰岛素样生长因子结合蛋白(IGFBP3)含量,应用荧光定量RTPCR法检测各组胎鼠脑组织IGFⅠmRNA表达。结果：与对照组相比较,模型组胎鼠脑组织中IGFⅠ(0.789±0.062ng/mgvs0.947±0.042ng/mg)、IGFⅡ(0.270±0.020ng/mgvs0.374±0.015ng/mg)含量均比对照组明显降低,IGFBP3(0.253±0.011ng/mgvs0.089±0.015ng/mg)含量比对照组明显升高,IGFⅠmRNA表达量(13.12±1.39)×104cps/μgRNAvs(21.28±3.54)×104cps/μgRNA比对照组明显降低,差异均有显著性(P<0.01)。与模型组相比较,小剂量和大剂量L精氨酸防治组IGFⅠ含量明显增高,分别为0.937±0.067ng/mg和0.858±0.077ng/mg,IGFⅡ含量明显增高,分别为0.318±0.018ng/mg和0.354±0.021ng/mg,IGFBP3含量明显降低,分别为0.132±0.006ng/mg和0.146±0.009ng/mg差异有显著性(P<0.01或<0.05)。同时小剂量和大剂量L精氨酸防治组IGFⅠmRNA表达量也明显增高,分别为(19.24±2.48)×104cps/μgRNA和(17.35±2.30)×104cps/μgRNAvs(13.12±1.39)×104cps/μgRNA,差异均有显著性(P<0.01)。结论：L精氨酸可增加被动吸烟致宫内发育迟缓胎鼠脑内IGFⅠ、IGFⅡ含量和IGFⅠmRNA的表达,降低IGFBP3含量。L精氨酸防治IUGR的机制与其对胰岛素样生长因子及其结合蛋白表达的影响有关。

Abstract:

OBJECTIVE: Intrauterine growth retardation (IUGR) may contribute to the disorder of development of fetal brains. L-arginine has been known to be effective in blood vessel distension and improving the blood circulation of placentas. Recent studies have shown that L-arginine can ameliorate the placental hypoxia and improve the development of fetus. This study aimed to explore the effects of L-arginine on the expression of insulin-like growth factor (IGF)-I, IGF-II,IGF binding protein-3(IGFBP3)and IGF-I mRNA in brains of IUGR rats and the possible mechanisms of L-arginine. METHODS: Thirty-six pregnant rats were randomly assigned into four groups: Control, Model, Low dose L-arginine (100 mg/kg) and High-dose L-arginine (200 mg/kg L-arginine) groups (n=9 each). IUGR was induced by passive smoking in rats from the last three groups. L-arginine was administered for the last two groups between days 8 and 20 of gestation. On day 21 of gestation, the pup rats were delivered by cesarean section. The levels of IGF-I, IGF-II and IGFBP3 in the brains of pup rats were measured by enzyme-linked immunoadsordent assay (ELISA) and the expression of IGF-I mRNA was detected by fluorescence quantitative PCR (FQ-PCR). RESULTS: The levels of IGF-I, IGF-II and IGF-I mRNA expression in the Model group were significantly lower than in the Control group, with the IGF-I levels of 0.789± 0.062 ng/mg vs 0.947±0.042 ng/mg, the IGF-II levels of 0.270±0.020 ng/mg vs 0.374±0.015 ng/mg and the IGF-I mRNA expression of (13.12±1.39)×104 cps/μg RNA vs (21.28±3.54) ×104 cps/μg RNA(P<0.01). In contrast, the IGFBP3 levels in the Model group were significantly higher than in the Control group (0.253±0.011 ng/mg vs 0.089± 0.015 ng/mg; P<0.01). Low or high dose L-arginine treatment increased significantly the IGF-I levels from 0.789± 0.062 ng/mg (Model group) to 0.937±0.067 ng/mg (low dose group) or 0.858±0.077 ng/mg (high dose group), the IGF-II levels from 0.270±0.020 ng/mg (Model group) to 0.318±0.018 ng/mg (low dose group) or 0.354±0.021 ng/mg (high dose group) and the IGF-I mRNA expression from (13.12±1.39) ×104 cps/μg RNA (Model group) to (19.24±2.48)×104 cps/μg RNA (low dose group) or (17.35±2.30)×104 cps/μg RNA (high dose group) (P< 0.01). The IGFBP3 levels were significantly reduced after low or high dose L-arginine treatment (0.132±0.006 ng/mg or 0.146±0.009 ng/mg) compared with those of the Model group (0.253±0.011 ng/mg) ( P<0.01). CONCLUSIONS: L-arginine can increase the levels of IGF-I and IGF-II and the IGF-I mRNA expression, and decrease the IGFBP3 level in the brain of rats with IUGR induced by passive smoking, thereby offering protective effects against IUGR.