芒柄花素对妊娠期糖尿病大鼠氧化应激损伤的影响

doi:10.11958/20221450

摘要/Abstract

摘要：

目的探究芒柄花素调节核因子E2相关因子2（Nrf2）/血红素单加氧酶-1（HO-1）/磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1（NQO1）信号通路对妊娠糖尿病（GDM）大鼠氧化应激损伤的影响。方法取SD孕鼠于其腹腔内注射链脲佐菌素诱导建立GDM模型，随机分为模型组、芒柄花素低剂量（50 mg/kg）组、芒柄花素高剂量（100 mg/kg）组、ML385（Nrf2抑制剂，20 mg/kg）组、芒柄花素高剂量+ML385组，每组9只，另取9只正常妊娠大鼠设为对照组。以芒柄花素和ML385分组处理后，检测各组大鼠体质量、血清空腹血糖（FSG）、总胆固醇（TC）、三酰甘油（TG）水平，胚胎存活率、胎鼠体质量、胎盘整体形态及超微结构，血清及胎盘组织丙二醛（MDA）、谷胱甘肽（GSH）、超氧化物歧化酶（SOD）和总抗氧化能力（TAC），胎盘组织Nrf2/HO-1/NQO1通路蛋白表达水平。结果与对照组比较，模型组大鼠胎盘超微结构发生损伤，胚胎存活率下降，血清及胎盘组织GSH、SOD、TAC水平降低，胎盘组织Nrf2、HO-1、NQO1蛋白表达降低，大鼠体质量、FSG、TG、TC、胎鼠体质量升高，血清及胎盘组织MDA水平升高（P<0.05）。与模型组比较，芒柄花素低剂量组、芒柄花素高剂量组大鼠胎盘超微结构损伤均减轻，胚胎存活率升高，血清及胎盘组织GSH、SOD、TAC水平升高，胎盘组织Nrf2、HO-1、NQO1蛋白表达均升高，大鼠体质量、FSG、TG、TC、胎鼠体质量降低，血清及胎盘组织MDA水平均降低（P<0.05），且高剂量芒柄花素作用更强。芒柄花素可逆转ML385对GDM大鼠的氧化应激损伤，改善大鼠胎盘损伤及不良妊娠结局。结论芒柄花素可通过激活Nrf2/HO-1/NQO1通路，从而减轻GDM大鼠氧化应激损伤，进而改善大鼠胎盘损伤及不良妊娠的结局。

关键词: 糖尿病，妊娠, 氧化性应激, 芒柄花素, 核因子E2相关因子2, 血红素单加氧酶-1, 磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1

Abstract:

Objective To explore the influence of formononetin on oxidative stress injury in gestational diabetes mellitus (GDM) rats by regulating nuclear factor erythroid-2-related factor 2 (Nrf2/hemeoxygenase-1 (HO-1)/NADPH quinone oxidoreductase-1 (NQO1) signaling pathway. Methods SD pregnant rats were injected intraperitoneally with streptozotocin to induce GDM model. Model rats were randomly grouped into the model group, the low-dose formononetin (50 mg/kg) group, the high-dose formononetin (100 mg/kg) group, the ML385 (Nrf2 inhibitor, 20 mg/kg) group, and the high-dose formononetin (100 mg/kg) + ML385 (20 mg/kg) group, 9 rats per group. Another 9 normal pregnant rats were regarded as the control group. After treatment with formononetin and ML385, body weight, serum fasting blood glucose (FSG), total cholesterol (TC), triglyceride (TG) levels, embryo survival rate, fetal body weight, the overall morphology and ultrastructure of placenta, levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and total antioxidant capacity (TAC) in serum and placental tissue were detected in rats of each group. The protein expression of Nrf2/HO-1/NQO1 pathway in placental tissue of rats was also detected in each group. Results Compared with the control group, the placental ultrastructure was damaged in the model group, and the embryonic survival rate, serum and placental tissue GSH, SOD, TAC levels, placental tissue Nrf2, HO-1, NQO1 protein expressions were significantly decreased (P<0.05). Body weight, FSG, TG, TC, fetal body weight, serum and placental tissue MDA levels were significantly increased in the model group (P<0.05). Compared with the model group, symptoms of placental ultrastructural damage were alleviated in the low-dose formononetin group and the high-dose formononetin group, and the embryonic survival rate, serum and placental tissue GSH, SOD, TAC levels, placental tissue Nrf2, HO-1, NQO1 protein expressions were all increased (P<0.05). The body weight, FSG, TG, TC, fetal body weight, serum and placental tissue MDA levels were all decreased (P<0.05), and the effect of high dose of formononetin was stronger. Changes of all indexes in the ML385 group were contrary to those in the formononetin treatment group, and formononetin can reverse the oxidative stress damage of ML385 on GDM rats, improve the placental damage and adverse pregnancy outcome of rats. Conclusion Formononetin can reduce oxidative stress injury in GDM rats by activating Nrf2/HO-1/NQO1 pathway, thereby improving placental damage and adverse pregnancy outcomes in rats.

Key words: diabetes, gestational, oxidative stress, anthocyanin, nuclear factor E2 related factor 2, heme monooxygenase-1, phosphatamide adenine dinucleotide quinone oxidoreductase-1

中图分类号:

R285.5

图/表 6

参考文献 20

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组别	母体体质量/g	FSG/ （mmol/L）	TG/ （mmol/L）	TC/ （mmol/L）
对照组	306.85±6.20	3.54±0.38	0.75±0.08	2.70±0.24
模型组	356.34±7.14^a	17.42±3.13^a	1.48±0.14^a	5.15±0.56^a
芒柄花素低剂量组	332.68±5.25^b	10.83±2.34^b	1.13±0.10^b	3.89±0.40^b
芒柄花素高剂量组	310.02±4.39^bc	4.02±0.54^bc	0.81±0.09^bc	2.78±0.29^bc
ML385组	372.69±7.81^b	26.79±3.48^b	1.95±0.19^b	6.26±0.48^b
芒柄花素高剂量+ ML385组	351.73±6.92^d	15.21±2.95^d	1.43±0.12^d	5.08±0.41^d
F	154.591^＊＊	115.158^＊＊	117.919^＊＊	108.771^＊＊

组别	母体体质量/g	FSG/ （mmol/L）	TG/ （mmol/L）	TC/ （mmol/L）
对照组	306.85±6.20	3.54±0.38	0.75±0.08	2.70±0.24
模型组	356.34±7.14^a	17.42±3.13^a	1.48±0.14^a	5.15±0.56^a
芒柄花素低剂量组	332.68±5.25^b	10.83±2.34^b	1.13±0.10^b	3.89±0.40^b
芒柄花素高剂量组	310.02±4.39^bc	4.02±0.54^bc	0.81±0.09^bc	2.78±0.29^bc
ML385组	372.69±7.81^b	26.79±3.48^b	1.95±0.19^b	6.26±0.48^b
芒柄花素高剂量+ ML385组	351.73±6.92^d	15.21±2.95^d	1.43±0.12^d	5.08±0.41^d
F	154.591^＊＊	115.158^＊＊	117.919^＊＊	108.771^＊＊

组别	胚胎存活率/%	胎鼠体质量/g
对照组	98.87±1.12	4.14±0.18
模型组	73.12±4.80^a	5.81±0.33^a
芒柄花素低剂量组	83.84±2.15^b	5.04±0.23^b
芒柄花素高剂量组	95.03±1.96^bc	4.25±0.14^bc
ML385组	64.01±2.07^b	6.53±0.29^b
芒柄花素高剂量+ML385组	75.74±3.39^d	5.74±0.21^d
F	200.071^＊＊	141.104^＊＊

组别	胚胎存活率/%	胎鼠体质量/g
对照组	98.87±1.12	4.14±0.18
模型组	73.12±4.80^a	5.81±0.33^a
芒柄花素低剂量组	83.84±2.15^b	5.04±0.23^b
芒柄花素高剂量组	95.03±1.96^bc	4.25±0.14^bc
ML385组	64.01±2.07^b	6.53±0.29^b
芒柄花素高剂量+ML385组	75.74±3.39^d	5.74±0.21^d
F	200.071^＊＊	141.104^＊＊

组别	MDA/ （nmol/L）	GSH/ （U/L）	SOD/ （U/L）	TAC/ （μmol/L）
对照组	2.81±0.32	64.62±5.31	91.12±7.54	15.21±1.43
模型组	4.93±0.45^a	40.85±3.08^a	60.78±4.65^a	8.16±0.64^a
芒柄花素低剂量组	3.87±0.38^b	49.97±3.53^b	73.89±5.31^b	11.08±0.91^b
芒柄花素高剂量组	2.95±0.31^bc	60.11±4.12^bc	88.67±6.24^bc	14.74±1.30^bc
ML385组	6.01±0.48^b	31.53±1.85^b	48.93±3.86^b	4.85±0.46^b
芒柄花素高剂量+ ML385组	4.82±0.43^d	42.04±2.98^d	63.51±4.38^d	8.74±0.65^d
F	87.584^＊＊	106.674^＊＊	82.515^＊＊	156.001^＊＊