Cui Wei-Wei, Peng Yun-Ru, Ding Yong-Fang
Jiangsu Province Hospital of Integrated of Chinese and Western Medicine Nanjing 210028,China Jiangsu Province Academy of Traditional Chinese Medicine Nanjing 210028,China.
Zhongguo Zhong Yao Za Zhi. 2019 Jul;44(14):2960-2965. doi: 10.19540/j.cnki.cjcmm.20190426.401.
The study aimed to investigate the mechanism of hepatoprotective effect of C-21 steroidal glucosides from Cynanchum auriculatum( Baishouwu) on oxidative stress in mice with liver injury. Mice were randomly divided into normal group,model group,positive control group,Baishouwu high group and Baishouwu low group. The liver injury model was induced by intraperitoneal injection of CCl4 peanut oil solution. All mice were sacrificed to collect blood and liver specimens. The activities of serum levels of ALT and AST were detected. The content of MDA and the activity of SOD in liver homogenate were examined by colorimetry method. Tissues were stained with hematoxylin-eosin for histological examination. The hepatic protein expressions of NF-κB p65,p-IκBα,i NOS and COX-2 were detected by Western blot. The mRNA expressions of TNF-α and IL-6 were determined by RT-PCR. It was found that treatment with C-21 steroidal glucosides from Baishouwu successfully attenuated liver injury induced by CCl4,as shown by decreased levels of serum biochemical indicators( AST,ALT)( P<0. 01). Administration of total C-21 steroidal glucosides enhanced the activity of SOD( P<0. 01) and decreased the content of MDA( P<0. 01) in liver homogenate. Microscopic features suggested that treatment with C-21 steroidal glucosides from Baishouwu was effective in inhibiting CCl4-induced hepatocyte edema and degeneration. Further studies showed that NF-κB p65 overexpression induced by CCl4 was decreased by C-21 steroidal glucosides,leading to the markedly down-regulated protein expression levels of p-IκBα,i NOS and COX-2,as well as the depression of TNF-α and IL-6 mRNA expressions. In conclusion,total C-21 steroidal glucosides from Baishouwu exhibited potent effect on oxidative stress pathway in mice with liver injury induced by CCl4,with enhanced activity of SOD,decreased content of MDA,and down-regulated levels of NF-κB p65,p-IκBα,i NOS and COX-2.
本研究旨在探讨白首乌C-21甾体苷对肝损伤小鼠氧化应激的肝保护作用机制。将小鼠随机分为正常组、模型组、阳性对照组、白首乌高剂量组和白首乌低剂量组。通过腹腔注射四氯化碳花生油溶液诱导肝损伤模型。处死所有小鼠以采集血液和肝脏标本。检测血清中谷丙转氨酶（ALT）和谷草转氨酶（AST）的活性。采用比色法检测肝匀浆中丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性。用苏木精-伊红染色进行组织学检查。通过蛋白质免疫印迹法检测肝组织中核因子-κB p65（NF-κB p65）、磷酸化IκBα（p-IκBα）、诱导型一氧化氮合酶（i NOS）和环氧化酶-2（COX-2）的蛋白表达。通过逆转录-聚合酶链反应（RT-PCR）测定肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的mRNA表达。结果发现，白首乌C-21甾体苷治疗成功减轻了四氯化碳诱导的肝损伤，血清生化指标（AST、ALT）水平降低（P<0.01）。给予总C-21甾体苷可增强肝匀浆中SOD活性（P<0.01）并降低MDA含量（P<0.01）。显微镜检查结果表明，白首乌C-21甾体苷治疗可有效抑制四氯化碳诱导的肝细胞水肿和变性。进一步研究表明，C-21甾体苷可降低四氯化碳诱导的NF-κB p65过表达，导致p-IκBα、i NOS和COX-2蛋白表达水平显著下调，以及TNF-α和IL-6 mRNA表达降低。总之，白首乌总C-21甾体苷对四氯化碳诱导的肝损伤小鼠的氧化应激途径具有显著作用，可增强SOD活性，降低MDA含量，并下调NF-κB p65、p-IκBα、i NOS和COX-2水平。
