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目的研究中药复方加味四逆散(JWSNS)对皮质酮和谷氨酸致损伤的PC12细胞中cAMP反应元件结合蛋白(CREB)及其磷酸化的影响。方法以200μmol.L-1Cort和50μmol.L-1Glu损伤的PC12细胞作为体外药理学研究模型,制备JWSNS含药血清,采用免疫组织化学法和West-ernblot法检测PC12细胞损伤模型中CREB和磷酸化CREB蛋白的表达。结果200μmol.L-1Cort和50μmol.L-1Glu可导致PC12细胞中CREB和p-CREB表达下降(CREB阳性细胞率分别为17.1%±4.2%和20.8%±5.7%,p-CREB表达量分别为0.587±0.123和0.515±0.157,P<0.05或P<0.01),10%JWSNS含药血清能升高CREB和p-CREB的表达(CREB阳性细胞率分别为62.6%±11.7%和79.5%±7.6%,p-CREB表达量分别为1.298±0.112和1.269±0.128,P<0.05或P<0.01)。结论10%JWSNS含药血清能通过调节cAMP-CREB信号通路发挥其神经保护和抗抑郁的作用。
Objective To investigate the effect of JWSNS on cAMP response element binding protein (CREB) and its phosphorylation in PC12 cells injured by corticosterone and glutamate. Methods PC12 cells damaged by 200μmol·L-1 Cort and 50μmol.L-1Glu were used as in vitro pharmacological models to prepare JWSNS-containing serum. Immunohistochemistry and West-ernblot method were used to detect CREB and phosphorylation in PC12 cells CREB protein expression. Results 200μmol.L-1Cort and 50μmol.L-1Glu decreased the expression of CREB and p-CREB in PC12 cells (CREB positive cells were 17.1% ± 4.2% and 20.8% ± 5.7%, respectively. The expression of p-CREB was 0.587 ± 0.123 and 0.515 ± 0.157, P <0.05 or P <0.01). The drug-containing serum of 10% JWSNS increased the expression of CREB and p-CREB (CREB positive cells were 62.6% ± 11.7% and 79.5% ± 7.6 %, p-CREB expression were 1.298 ± 0.112 and 1.269 ± 0.128 respectively, P <0.05 or P <0.01). Conclusion 10% JWSNS drug-containing serum can exert its neuroprotective and anti-depressant effects by regulating the cAMP-CREB signaling pathway.