Taicang Traditional Chinese Medicine Hospital, Affiliated to Nanjing University of Chinese Medicine, Taicang 215400, China.
Department of Gynaecology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China.
Biomed Res Int. 2020 Dec 1;2020:5840967. doi: 10.1155/2020/5840967. eCollection 2020.
Ge-Gen decoction (GGD) is widely used for the treatment of primary dysmenorrhea (PD) in China. However, the mechanisms that underlie this effect are unclear. We investigated the protective mechanism of GGD in a rat model of PD using label-free quantitative proteomics. The model was established by the administration of estradiol benzoate and oxytocin. Thirty rats were divided into three groups (ten rats/group): a control group (normal rats), a model group (PD rats), and a treatment group (PD rats treated with GGD). The serum levels of prostaglandin E2 (PGE2) and prosn F2 (PGF2) were measured by ELISA. Nanohigh-performance liquid chromatography-tandem mass spectrometry (nano-HPLC-MS/MS) was used to identify differentially expressed proteins (DEPs), and bioinformatics was used to investigate the protein function. Proteomic data were validated by western blot analysis. Oxytocin-induced writhing responses and abnormal serum levels of PGE2 and PGF2 were reversed following the administration of GGD. A total of 379 DEPs were identified; 276 were identified between the control group and the model group, 144 were identified between the model group and the treatment group, and 41 were identified as DEPs that were common to all groups. Bioinformatics revealed that the DEPs between the control group and the model group were mainly associated with cellular component biogenesis and binding processes. The DEPs between the model group and the treatment group were mainly involved in the protein binding and metabolic process. The expression levels of HSP90AB1 and the phosphorylation levels of ERK, JNK, and P-p38 in the uteri of rats in the three groups were consistent with the proteomic findings; MAP kinases (ERK, JNK, and p38) are known to be involved in the production of inflammatory cytokines and oxytocin signaling while HSP90AB1 is known to be associated with estrogen signaling. Collectively, these data indicate that GGD may exert its protective function on PD by regulating the inflammatory response and signaling pathways associated with oxytocin and estrogen.
葛根汤（GGD）广泛用于中国原发性痛经（PD）的治疗。然而，其作用机制尚不清楚。我们使用无标记定量蛋白质组学研究了 GGD 在 PD 大鼠模型中的保护机制。该模型通过苯甲酸雌二醇和催产素给药建立。将 30 只大鼠分为三组（每组 10 只）：对照组（正常大鼠）、模型组（PD 大鼠）和治疗组（PD 大鼠用 GGD 治疗）。通过 ELISA 测定血清前列腺素 E2（PGE2）和前列腺素 F2（PGF2）水平。纳米高效液相色谱-串联质谱（nano-HPLC-MS/MS）用于鉴定差异表达蛋白（DEPs），并进行蛋白质功能的生物信息学分析。通过 Western blot 分析验证蛋白质组学数据。葛根汤给药后可逆转催产素诱导的扭体反应及 PGE2 和 PGF2 血清水平异常。共鉴定出 379 个 DEPs；其中 276 个在对照组和模型组之间鉴定，144 个在模型组和治疗组之间鉴定，41 个在所有组中鉴定为 DEPs。生物信息学显示，对照组和模型组之间的 DEPs 主要与细胞成分生物发生和结合过程有关。模型组和治疗组之间的 DEPs 主要参与蛋白质结合和代谢过程。三组大鼠子宫中 HSP90AB1 的表达水平和 ERK、JNK 和 P-p38 的磷酸化水平与蛋白质组学结果一致；MAP 激酶（ERK、JNK 和 p38）参与炎症细胞因子和催产素信号的产生，而 HSP90AB1 与雌激素信号有关。综上所述，这些数据表明，葛根汤可能通过调节与催产素和雌激素相关的炎症反应和信号通路对 PD 发挥保护作用。
