黄芩苷通过PI3K/Akt/mTOR通路参与子痫前期 大鼠免疫状态及炎症反应的调控及机制

目的：研究黄芩苷对子痫前期大鼠免疫状态及炎症反应的调控机制及PI3K/Akt/mTOR 信号通路的影响。 方法：雌性大鼠随机分为对照组、模型组和处理组。处理组和模型组大鼠妊娠13~21 d 时给予L- 精氨酸甲酯，治 疗组大鼠于妊娠17~21 d 时给予黄芩苷注射液，对照组和模型组大鼠具给予等量的生理盐水，3 组均持续给药至 妊娠21 d。妊娠21 d 时，采用电脑尾袖式血压计测定各组大鼠血压；采用全自动生化分析仪测定各组大鼠血清中 尿素氮（BUN）水平；测定大鼠血清中和胎盘中炎症因子白细胞介素（IL）-6、IL-1β 和肿瘤坏死因子-α（TNF-α） 水平；采用蛋白免疫印迹法测定大鼠胎盘组织中Akt 和mTOR 蛋白含量、尿蛋白水平和炎症介质NF-κB和IκB-α 蛋白表达水平；采用RT-qPCR 法测定大鼠胎盘组织中α7nAChR RNA表达水平；采用流式细胞仪测定各组大鼠全 血中T 淋巴细胞的比例。结果：模型组大鼠舒张压、收缩压、BUN和尿蛋白水平显著高于对照组，处理组舒张 压、收缩压、BUN和尿蛋白水平与模型组相比均显著降低；模型组大鼠IL-6、IL-1β 和TNF-α 水平显著高于对照 组，处理组IL-6、IL-1β 和TNF-α 水平与模型组相比均显著降低；模型组大鼠Akt 和mTOR 蛋白表达水平显著低 于对照组，处理组Akt 和mTOR 蛋白表达水平与模型组相比均显著升高；模型组大鼠NF-κB和IκB-α 蛋白表达水 平显著高于对照组，处理组NF-κB和IκB-α 蛋白表达水平与模型组相比均显著降低；模型组与对照组胎盘组织中 α7nAChR RNA表达水平相当，处理组大鼠胎盘组织中α7nAChR RNA表达水平显著高于模型组和对照组；模型 组大鼠CD3+、CD4+、CD4+/CD8+ 水平均显著高于对照组，CD8+ 水平显著低于对照组；处理组大鼠与模型组相比， CD3+、CD4+、CD4+/CD8+ 水平均显著降低，CD8+ 水平显著升高。结论：黄芩苷可以通过促进PI3K/Akt/mTOR 信 号通路抑制机体炎症反应，改善机体免疫功能。

Regulation of immune status and inflammatory response in pre-eclamptic rats by baicalin through PI3K/Akt/mTOR pathway and its mechanism

Objective To explore the baicalin mechanism of regulation on the immune status and inflammatory response preeclamptic rats through PI3K/Akt/mTOR pathway. Methods Thirty-six female rats were randomly divided into a control group， a model group， and a treatment group. The rats in the treatment group and the model group were given L-arginine methyl ester at 13-21 days of pregnancy， and the rats in the control group were injected subcutaneously with the same amount of normal saline for continuous administration until the 21st day of pregnancy. The rats in the treatment group were given baicalin injection from 17 to 21 days of gestation. The rats in the control group and the model group were given the same amount of normal saline. The three groups continued to be administered until 21 days of gestation. On the 21st day of pregnancy， the blood pressure of rats in each group was measured with a computer tail-cuff sphygmomanometer. The automatic biochemical analyzer was used to determine the serum urea nitrogen（ BUN） level of rats in each group. The levels of inflammatory factors interleukin （IL）-6， IL-1β and tumor necrosis factor-α（ TNF-α） in serum and placenta of rats were measured. The contents of Akt and mTOR protein， urinary protein level and the expression levels of inflammatory mediators NF-κB and IκB-α protein in rat placenta were determined by Western blotting. RT-qPCR method was used to determine the expression level of α7nAChR RNA in rat placenta. The proportion of T lymphocytes in the whole blood of rats in each group was determined by flow cytometry. Results The diastolic blood pressure， systolic blood pressure， BUN， and urine protein levels of the model group weresignificantly higher than those of the control group. The diastolic blood pressure， systolic blood pressure， BUN， and urine protein levels of the treatment group were significantly lower than those of the model group. The levels of IL-6， IL-1β and TNF-α in the model group were signi ficantly higher than those in the control group ， and the levels of IL-6， IL-1β and TNF-α in the treatment group were significantly lower than those in the model group. The expression levels of Akt and mTOR proteins in the model group were significantly lower than those in the control group ， and the expression levels of Akt and mTOR proteins in the treatment group were significantly higher than those in the model group. The protein expression levels of NF-κB and IκB-α in the model group were significantly higher than those in the control group， and the expression levels of NF-κB and IκB-α in the treatment group were significantly lower than those in the model group. The expression level of α7nAChR RNA in the placenta tissue of the model group and the control group was equivalent， and the expression level of α7nAChR RNA in the placenta tissue of the treatment group was significantly higher than that of the model group and the control group. The CD3+， CD4+， CD4+/CD8+ levels of the model group were significantly higher than those of the control group， and the CD8+ level was significantly lower than the control group. Compared with the model group， the rats in the treatment group had significantly lower CD3+， CD4+， CD4+/CD8+ levels， and significantly increased CD8+ levels. Conclusion Baicalin can inhibit the body’s inflammatory response and improve the body’s immune function by promoting the PI3K/Akt/ mTOR signal pathway in rat.