左归丸对成骨细胞氧化应激损伤的保护作用

Zuogui Pill has protective effect against oxidative stress injury in osteoblasts

背景:最新研究发现氧化应激在骨质疏松症中发挥着重要作用,衰老、雌激素缺乏、核因子E2相关因子2(Nrf2)、血红素加氧酶1与活性氧的产生和骨质疏松发病有着重要的关系。目的:分析左归丸对成骨细胞氧化应激损伤的保护作用机制。方法:实验方案经哈尔滨医科大学附属第二医院伦理委员会批准。分离培养SD乳鼠颅骨成骨细胞,实验分为空白组、模型组(300μmol/L H2O2)、左归丸组(300μmol/L H2O2+10%左归丸含药血清)。用H2O2建立成骨细胞氧化应激损伤模型;在成骨细胞氧化应激损伤同时,用左归丸含药血清培养成骨细胞,空白组不做任何处理。测定各组成骨细胞中丙二醛和超氧化物歧化酶含量;Western blot检测成骨细胞中核因子E2相关因子2和血红素加氧酶1蛋白表达。结果与结论:①与空白组比较,模型组中丙二醛含量明显升高,超氧化物歧化酶的含量明显降低;与模型组比较,左归丸组中丙二醛含量明显降低,超氧化物歧化酶的含量明显升高;②与空白组比较,模型组成骨细胞中血红素加氧酶1和核因子E2相关因子2表达明显增加;左归丸组成骨细胞中血红素加氧酶1和核因子E2相关因子2表达明显高于模型组;③结果说明,左归丸对成骨细胞氧化损伤具有保护作用。

BACKGROUND:Recent studies have found that oxidative stress plays an important role in osteoporosis.Osteoporosis is closely related to aging,estrogen deficiency,nuclear factor E2 related factor 2,heme oxygenase 1 and reactive oxygen species production.OBJECTIVE:To explore the protective mechanism of Zuogui Pill on oxidative stress injury of osteoblasts.METHODS:The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Harbin Medical University.Osteoblasts were isolated and cultured from the skull of neonatal Sprague-Dawley rats,and were divided into three groups:control group,model group and Zuogui Pill group.The oxidative stress injury model of osteoblasts was established with 300μmol/L H2O2 in the model and Zuogui Pill groups.Osteoblasts in the Zuogui Pill group were cultured with Zuogui Pill containing serum,and the control group did not undertake any treatment.Levels of malondialdehyde and superoxide dismutase in osteoblasts were determined.The expression of nuclear factor E2 related factor 2 and heme oxygenase 1 protein in osteoblasts was detected by western blot.RESULTS AND CONCLUSION:Compared with the control group,the content of malondialdehyde increased significantly,and the level of superoxide dismutase decreased significantly in the model group.Compared with the model group,the content of malondialdehyde decreased significantly and the level of superoxide dismutase increased significantly in the Zuogui Pill group.Western blot results showed that nuclear factor E2 related factor 2 and heme oxygenase 1 expression in osteoblasts of the model group increased significantly compared with the control group,and the expression of nuclear factor E2 related factor 2 and heme oxygenase 1 in osteoblasts of Zuogui Pill group was significantly higher than that of the model group.To conclude,Zuogui Pill can protect osteoblasts from oxidative stress injury.