芍药苷调控环磷酸腺苷 /蛋白激酶 A/环磷酸腺苷反应元件结合蛋白通路对脑卒中大鼠的影响

目的探讨芍药苷调控环磷酸腺苷( cAMP)/蛋白激酶 A(PKA)/环磷酸腺苷反应元件结合蛋白( CREB)通路对脑卒中大鼠的影响。方法该研究起止时间为 2019年 12月至 2020年 12月。 40只 SD大鼠按随机数字表法分为假手术组、模型组、药苷组(灌胃 30 mg/kg芍药苷)、 H89组［腹腔注射 1 mg/kg H89(PKA抑制剂)］、芍药苷 +H89组(灌胃芍药苷后腹腔注射 H89)芍，除假手术组仅进行动脉分离，其余各组均构建缺血性脑卒中模型，模型构建成功后进行药物干预，持续给药 2周，模型组、假手术组以生理盐水代替。根据 Zea Longa评分法对各组大鼠进行神经功能缺损评分， Morris水迷宫测定大鼠记忆行为学能力，酶联免疫吸附测定( ELISA)试剂盒检测大鼠血清炎性因子水平，苏木精 -伊红( HE)染色观察大鼠脑组织病理学变化，蛋白质印迹法检测大鼠脑组织 cAMP/PKA/CREB通路相关蛋白表达。结果与假手术组相比，模型组脑组织受损严重，大鼠神经功能缺损评分、血清中炎性因子水平显著升高( P<0.05)记忆行为学能力、脑组织中 cAMP［( 0.31±0.05)比( 1.03±0.23)］、 PKA［( 0.30±

The effect of paeoniflorin on stroke rats by regulating cAMP/PKA/CREB pathway

Objective To explore the effect of paeoniflorin on stroke rats by regulating cyclic adenosine monophosphate/protein kinase A/cyclic adenosine monophosphate response element binding protein (cAMP/PKA/CREB) pathway.Methods The study startedfrom December 2019 and ended in December 2020. Forty SD rats were assigned into sham operation group, model group, paeonifloringroup (gavage of 30 mg/kg paeoniflorin), H89 group intraperitoneal injection of 1 mg/kg H89 (PKA inhibitor)], paeoniflorin+ H89group (gavage of paeoniflorin followed by intraperitoneal injection of H89) according to random number table method. Only the sham operation group performed arterial separation, while the other groups all established models of ischemic stroke, and after the model wassuccessfully constructed, drug intervention was carried out for 2 weeks. The model group and sham operation group were given normalsaline instead. The Zea Longa scoring method was used to score the rats in each group for neurological deficits, Morris water maze wasused to measure the memory behavior ability of rats, the enzyme-linked immunosorbent assay (ELISA) kit the level of serum inflammatory factors in rats, hematoxylin -eosin (HE) staining the pathological changes of rat brain tissue, and Western blotting the expression ofcAMP/PKA/CREB pathway related proteins in rat brain tissue.Results Compared with the sham operation group, the brain tissue ofthe model group was severely damaged, the neurological deficit score and serum inflammatory factor level in the rats were significantlyincreased (P<0.05), the memory behavior ability and the protein expressions of cAMP (0.31±0.05) vs. (1.03±0.23)], PKA (0.30±0.06) vs. (1.05±0.24)],, phosphorylated CREB (0.29±0.08) vs. (1.02±0.23)] were significantly reduced in brain tissue (P<0.05). Comparedwith the model group, the brain tissue damage of rats in the paeoniflorin group was relieved, the neurological deficit score and serum inflammatory factor level were significantly reduced (P<0.05), the memory behavior ability and the protein expressions of cAMP (0.79± 0.20) vs. (0.31±0.05)], PKA (0.75±0.19) vs. (0.30±0.06)], and phosphorylated CREB (0.77±0.23) vs. (0.29±0.08)] were significantly increased (P<0.05). Compared with the paeoniflorin group, the brain tissue damage of rats in the paeoniflorin+H89 group was more severe, the neurological deficit score and serum inflammatory factor level were significantly increased (P<0.05), the memory behavior ability and the protein expressions of cAMP (0.60±0.13) vs. (0.79±0.20)], PKA (0.53±0.12) vs. (0.75±0.19)], and phosphorylated CREB (0.55±0.14) vs. (0.77±0.23)] were significantly reduced (P<0.05).Conclusion Paeoniflorin can protect the neurological function of stroke rats, which may be achieved by activating the cAMP/PKA/CREB pathway.