淫羊藿苷调控急性淋巴细胞白血病PI3K/Akt信号传导通路的实验研究  

Experimental Study on the Regulation of Icariin on PI3K/Akt Signal Transduction Pathway in Acute Lymphocytic Leukemia

作　　者：王黎 柯鸿 Wang Li;Ke Hong(The people＇s Hospital Affiliated to Hubei Medical College, Hubei ,Shiyan 442000, Chin)

机构地区：[1]湖北医药学院附属人民医院,湖北十堰442000

出　　处：《中国中医急症》2018年第6期1000-1003,1011,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：湖北省十堰市科技局科学技术研究与开发项目(17Y35)

摘　　要：目的观察淫羊藿苷对近交系DBA小鼠急性淋巴细胞白血病磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号传导通路的影响并分析其机制。方法近交系DBA/2小鼠60只,采用皮下注射急性淋巴细胞白血病细胞系L1210细胞以建立L1210白血病模型,然后随机均分为模型对照组、阳性对照组和淫羊藿苷2.5 mL/kg组、淫羊藿苷5 mL/kg组和淫羊藿苷10 mL/kg组(n=12只),1周后淫羊藿苷2.5 mL/kg组、5 mL/kg组和10 mL/kg组灌胃给予对应剂量的淫羊藿苷,模型对照组灌胃给予等体积0.9%氯化钠注射液对照,阳性对照组灌PI3K/Akt抑制剂LY294002对照。治疗3周后采用人工法计数尾静脉血细胞分类及血象;取小鼠肝、脾组织,采用Westem blot检测B细胞淋巴瘤/白血病-2(Bcl-2)及磷酸化蛋白激酶(p-Akt)蛋白表达,实时定量PCR(RT-PCR)检测组织同源性磷酸酶-张力蛋白(PTEN)m RNA和Akt m RNA的表达。结果与模型对照组比较,淫羊藿苷2.5 mL/kg组、5 mL/kg组和10 mL/kg组尾静脉血细胞淋巴细胞、白细胞和L1210细胞总数均明显降低,而中性粒细胞升高(P<0.05);小鼠肝、脾组织Bcl-2、p-Akt蛋白和Akt m RNA蛋白表达水平显著降低,PTEN蛋白和PTEN m RNA则高表达(P<0.05);且上述指标中,淫羊藿苷5 mL/kg组与2.5 mL/kg组比较(P<0.05);淫羊藿苷10 mL/kg与5 mL/kg组和2.5 mL/kg组比较,差异有统计学意义(P<0.05)。至治疗结束时淫羊藿苷2.5 mL/kg组、5 mL/kg组和10 mL/kg组小鼠存活数、体质量、肝脾和皮下瘤重与模型对照组比较(P<0.05),差异有统计学意义。结论淫羊藿苷可能通过阻断PI3K/Akt信号传导通路而实现抑制L1210细胞增殖效应,且这种作用呈剂量依赖性。Objective： To observe the effect of epimedium glycoside on signaling transcription pathways of phosphatidyl inositol 3 kinase（PI3 K）/protein kinase B（Akt） of inbred line DBA mice with acute lymphocytic leukemia and analyze the influence of its mechanism. Methods： 60 Inbred line DBA/2 mice were treated by subcutaneous injection of acute lymphocytic leukemia L1210 cell of cell line to set up L1210 leukemia model,and then they were randomly divided into the model control group,positive control group and 2.5 mL/kg group,5 mL/kg and 10 mL/kg group（n = 12）. After 1 week,2.5 mL/kg group,5 mL/kg group and 10 mL/kg group were gavaged according to the corresponding dose of epimedium glycoside;the model control group was treated by the same volume of saline as a comparision;positive control group was filled with PI3 K LY294002/Akt inhibitors. After 3 weeks of treatment,the blood cells were classified and hematosed with artificial method. Livers and spleen tissues of mice were detected by the Westem blot to test expressions of B-2 lymphocyte tumor（Bcl-2） and phosphorylated protein kinase（p-Akt） protein;real-time quantitative PCR（rt-pcr） was used to detect expressions of organization homologous protein phosphatase-tension（PTEN） m RNA and Akt m RNA. Results： Compared with the model control group,in 2.5 mL/kg group,5 mL/kg group and 10 mL/kg,tail venous blood cells,lymphocytes,white blood cells and L1210 cells were significantly lower but neutrophils was higher（P〈0.05）. The expression levels of bcl-2,p-Akt and Akt m RNA protein in mice liver and spleen tissues were significantly lower,while PTEN m RNA had high expression（P〈0.05）. In the indicators above,differences were statistically significant between 5 mL/kg group and 2.5 mL/kg group（P〈0.05）;differences were statistically significant among 10 mL/kg,5 mL/kg group and 2.5 mL/kg group（P〈0.05）. Differences were statistically significant in the survival numbers,weight,liver,spleen and subcutaneous tumor among the control g

关 键 词：急性淋巴细胞白血病 淫羊藿苷 PI3K/AKT信号传导通路 近交系DBA小鼠 

分 类 号：R285.5[医药卫生—中药学]