葛根素对妊娠高血压综合征大鼠的作用以及肾损伤的影响
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陕西省卫生健康科研基金项目(2022D067);陕西省重点研发计划(2022SF-408)


Effects of puerarin on hypertensive syndrome of pregnancy and renal injury in rats
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    摘要:

    目的 探究葛根素(PUE)在妊娠高血压综合征(PIH)模型大鼠中的作用以及对PIH所引发的肾损伤的影响。方法 采用皮下注射剂量为125 mg/kg的NG-硝基-L-精氨酸甲酯构建PIH大鼠模型,将成功妊娠的SD大鼠分为对照组、模型组、50 mg/kg葛根素组(50 PUE组)和100 mg/kg葛根素组(100 PUE组),其中模型组、50 PUE组和100 PUE组为PIH大鼠,每组10只。妊娠第16天,50 PUE组和100 PUE组大鼠每天分别给予剂量为50 mg/kg和100 mg/kg的葛根素灌胃处理,每天1次,连续处理5天;对照组和模型组大鼠同期均给予等量生理盐水灌胃。检测各组大鼠在妊娠第1、12、15、18和第21天的尾动脉血压;妊娠第20天,收集各组大鼠24 h内的尿液,采用Bradford法检测24 h尿蛋白水平。妊娠第21天,分离和收集各组大鼠血清和肾组织,采用自动生化分析仪检测血肌酐(SCr)和血清尿素氮(BUN)水平;采用ELISA法检测血清中IL-1β、IL-6和TNF-α的水平;采用试剂盒检测大鼠肾组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH)含量。采用HE染色和PAS染色检测大鼠肾组织病理改变;采用TUNEL染色检测大鼠肾组织细胞凋亡水平;采用免疫组织化学染色法和Western blot检测大鼠肾组织中Bcl-2、Bax和cleaved caspase-3蛋白表达量。结果 妊娠第15天,与对照组比较,模型组、50 PUE组和100 PUE组大鼠尾动脉血压均升高(P<0.05);妊娠第18天和妊娠第21天,与模型组比较,50 PUE组和100 PUE组大鼠尾动脉血压均降低(P<0.05)。妊娠第21天,与对照组比较,模型组大鼠肾组织出现严重胞质空泡化,肾小管水肿和扩张,糖原沉积;24 h尿蛋白以及血清中IL-1β、IL-6、TNF-α、SCr和BUN水平均升高(P<0.05);肾组织中SOD和GSH含量以及Bcl-2蛋白相对表达量均降低(P<0.05),MDA含量以及Bax和cleaved caspase-3蛋白相对表达量均升高(P<0.05),TUNEL阳性细胞率升高(P<0.05)。与模型组比较,50 PUE组和100 PUE组大鼠肾组织损伤程度均降低,24 h尿蛋白以及血清中IL-1β、IL-6、TNF-α、SCr和BUN水平均降低(P<0.05);肾组织中SOD和GSH含量以及Bcl-2蛋白相对表达量均升高(P<0.05),MDA含量以及Bax和cleaved caspase-3蛋白相对表达量均降低(P<0.05),TUNEL阳性细胞率降低(P<0.05)。结论 葛根素可改善PIH大鼠高血压症状,抑制肾脏炎症和氧化应激反应,抑制细胞凋亡,减轻PIH所引发的肾组织损伤

    Abstract:

    Objective To investigate the effects of puerarin (PUE) on the rats with pregnancy-induced hypertension syndrome (PIH), and the effects of PUE on renal injury induced by PIH. Methods The model of PIH rats were established by 〖HJ48x〗subcutaneous injection of NG-nitro-L-arginine methyl ester at the dose of 125 mg/kg. SD rats with successful pregnancy were divided into control group, model group, 50 mg/kg PUE group (50 PUE group) and 100 mg/kg PUE group (100 PUE group). The model group, 50 PUE group and 100 PUE group were PIH rats with 10 rats in each group. On the 16th day of pregnancy, rats in 50 PUE group and 100 PUE group were given PUE at the dose of 50 mg/kg and 100 mg/kg once a day, for 5 consecutive days. Rats in control group and model group were given the same amount of normal saline. The blood pressure of caudal artery was detected on the 1st, 12th, 15th, 18th and 21st days of pregnancy. On the 20th day of pregnancy, urine samples were collected within 24h of each group, and 24h urinary protein levels were detected by Bradford method. On the 21st day of pregnancy, serum and renal tissues of rats in each group were separated and collected. Serum creatinine (SCr) and serum urea nitrogen (BUN) levels were detected by automatic biochemical analyzer. The levels of IL-1β, IL-6 and TNF-α in serum were detected by ELISA. The contents of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH) in rat renal tissue were detected by a kit. The pathological changes of renal tissue were detected by HE and PAS staining. The apoptosis level of rat renal tissue was detected by TUNEL staining. The protein expression levels of Bcl-2, Bax and cleaved caspase-3 in rat renal tissues were detected by immunohistochemical staining and Western blot. Results On the 15th day of pregnancy, compared with the control group, the tail artery blood pressure in model group, 50 PUE group and 100 PUE group were increased (P<0.05). On the 18th and 21st days of pregnancy, compared with the model group, the tail artery blood pressure in 50 PUE group and 100 PUE group were decreased (P<0.05). On the 21st day of pregnancy, compared with the control group, severe cytoplasmic vacuolation, edema and dilation of renal tubules and glycogen deposition occurred in the model group. The level of 24h urinary protein and the levels of IL-1β, IL-6, TNF-α, SCr and BUN in serum were increased (P<0.05). The contents of SOD and GSH and the relative expression of Bcl-2 protein in renal tissues were decreased (P<0.05), MDA content and the relative expression of Bax and cleaved caspase-3 protein were increased (P<0.05), and the rate of TUNEL positive cells was increased (P<0.05). Compared with model group, the renal tissue injury degree of rats in 50 PUE group and 100 PUE group were decreased, and the levels of 24h urinary protein and the levels of IL-1β, IL-6, TNF-α, SCr and BUN in serum were decreased (P<0.05). The contents of SOD and GSH and the relative expression of Bcl-2 protein in renal tissues were increased (P<0.05), MDA content and the relative expression of Bax and cleaved caspase-3 protein were decreased (P<0.05), and the rate of TUNEL positive cells was decreased (P<0.05).Conclusion PUE can improve the symptoms of hypertension in PIH rats, inhibit renal inflammation and oxidative stress, inhibit cell apoptosis, and alleviate the renal tissue damage caused by PIH

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  • 在线发布日期: 2025-05-23
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