mTOR及S6K1在免疫介导再生障碍性贫血模型小鼠骨髓中的表达

Expression of m TOR and S6K1 in bone marrow of immune conductive aplastic anemia mice

目的通过检测再生障碍性贫血模型小鼠骨髓中脂肪调控因子哺乳动物雷帕霉素靶蛋白(m TOR)及通路下游核糖体蛋白激酶S6K1的表达,研究再生障碍性贫血小鼠骨髓脂肪化的机制。方法 (1)制备免疫介导再生障碍性贫血小鼠模型。(2)造模开始后观察记录小鼠状态并定期测小鼠血常规以验证造模是否成功。(3)免疫组织化学方法检测骨髓细胞m TOR及S6K1的表达情况。所有数据采用SPSS 22.0软件进行统计学分析,数据经正态性检验,两组均数的比较采用单样本t检验,相关分析采用Pearson相关性检验。结果 (1)再生障碍性贫血小鼠造模成功:与对照组相比,再生障碍性贫血组小鼠第5天开始出现精神状态不佳、皮毛光泽度降低、体质量下降,造模14 d后死亡5只。外周血细胞计数明显下降,表现为白细胞(WBC)、血红蛋白(Hb)、血小板(PLT)三系均减少,造模14 d时血常规分别为:对照组(10.52±0.73)×109/L,(132±6.3)g/L,(469±114)×109/L,再生障碍性贫血组(0.97±0.57)×109/L,(74±9.3)g/L,(57±32)×109/L(P均<0.001)。(2)免疫组化显示再生障碍性贫血小鼠骨髓m TOR及S6K1的表达明显高于对照组,分别表现为对照组74.84±6.33、63.45±5.38,再生障碍性贫血组113.27±16.23、108.56±12.77(P均<0.001)。(3)相关分析显示再生障碍性贫血小鼠m TOR及S6K1与WBC、Hb、PLT呈负相关,m TOR与S6K1呈正相关。结论 m TOR及S6K1在再生障碍性贫血模型小鼠骨髓细胞的表达明显增加,与血细胞变化呈负相关,m TOR及S6K1可能在骨髓脂肪细胞的形成中起一定作用即参与了再生障碍性贫血小鼠骨髓的脂肪化,继而可能抑制了骨髓造血。

Objective To research the mechanism of bone marrow fat of aplastic anemia mice, through detecting the expression of m TOR, a fat regulatory factor, and S6K1, which is located in m TOR pathway downstream. Methods(1) The success of immune conductive aplastic anemia mice was the key factor of the whole experiment.(2) Observed the growing status of the mice, and measured the hemogram regularly to verify the success of aplastic anemia mice model.(3) Detected the expression of m TOR and S6K1 in bone marrow of immune conductive aplastic anemia mice with immunohistochemistry. All data was checked with software SPSS 22.0. To check up the data of all groups with normal test. t-test of single sample was used in the comparison of means of two groups. Correlation analysis was done with Pearson correlation. Results(1) The AA mice model was successfully made: compared with the control group, on the fifth day of the preparation of immune conductive aplastic anemia mice, the AA group began to appear poor mental status, reduced gloss of skin and weight loss, and then 5 died after 14 days. Three series decreased in blood cell count of the AA group, that is to say, the counts of WBC, Hb, and PLT of the AA group decreased obviously than the control group, after 14 days blood count present as the control group(10.52±0.73)×109/L,(132±6.3)g/L,(469±114)×109/L, the AA group(0.97±0.57)×109/L,(74±9.3)g/L,(57±32)×109/L(all P<0.001).(2) Immunohistochemistry showed the expression of m TOR and S6K1 in the AA group was significantly higher than the control group, manifested as the control group 74.84±6.33, 63.45±5.3), the AA group 113.27±16.23, 108.56±12.77(all P<0.001).(3) Correlation analysis showed the expression of m TOR and S6K1 was negatively related with the count of WBC, Hb, and PLT in the AA group. In AA group m TOR and S6K1 were positively correlated. Conclusions Expression of m TOR and S6K1 in bone marrow cells of immune conductive aplastic anemia mice significantly increased and was negatively correlated with the count of WBC, Hb, and PLT. In conclusion, m TOR and S6K1 are probably involved in the formation of bone marrow fat of aplastic anemia mice, and then suppress bone marrow.