Li Mei-Yi, Zhang Yan-Bo, Zuo Huan, Liu Li-Li, Niu Jing-Zhong
Department of Neurology, Taishan Chronic Disease Hospital, Taian 271000, China.
Sheng Li Xue Bao. 2012 Feb 25;64(1):41-7.
The present study was to investigate the effect of Salvia miltiorrhiza Bunge. f. alba (SMA) pharmacological pretreatment on apoptosis of cultured hippocampal neurons from neonate rats under oxygen-glucose deprivation (OGD). Cultured hippocampal neurons were randomly divided into five groups (n = 6): normal plasma group, low dose SMA plasma (2.5%) group, middle dose SMA plasma (5%) group, high dose SMA plasma (10%) group and control group. The hippocampal neurons were cultured and treated with plasma from adult Wistar rats intragastrically administered with saline or aqueous extract of SMA. The apoptosis of neurons was induced by glucose-free Earle's solution containing 1 mmol/L Na2S2O4 and labeled by MTT and Annexin V/PI double staining. Moreover, protein expressions of Bcl-2 and Bax were detected by immunofluorescence. The results showed that few apoptotic cells were observed in control group, whereas the number of apoptotic cells was greatly increased in normal plasma group and low dose SMA plasma group. Both middle and high dose SMA plasma could protect cultured hippocampal neurons from apoptosis induced by OGD (P < 0.05). The protective effect of high dose SMA plasma was stronger than that of middle one (P < 0.05). Compared to control, normal plasma and low dose SMA plasma groups, middle and high dose SMA plasma groups both showed significantly higher levels of Bcl-2 (P < 0.05 or 0.01), whereas expressions of Bax was opposite. There were no significant differences of Bcl-2 and Bax expressions between middle and high dose SMA plasma groups. Number of Bcl-2- and Bax-positive cells had similar tendency. Bcl-2/Bax (number of positive cells) ratio was higher in high dose SMA plasma group than those of all the other groups (P < 0.05 or 0.01). These results suggest that pharmacological pretreatment of blood plasma containing middle and high dose SMA could raise viability and inhibit apoptosis of OGD-injured hippocampal neurons by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax.
本研究旨在探讨白花丹参（SMA）药理预处理对新生大鼠海马神经元在氧糖剥夺（OGD）条件下凋亡的影响。将培养的海马神经元随机分为五组（n = 6）：正常血浆组、低剂量SMA血浆（2.5%）组、中剂量SMA血浆（5%）组、高剂量SMA血浆（10%）组和对照组。用生理盐水或SMA水提取物灌胃成年Wistar大鼠后获取血浆，对海马神经元进行培养和处理。通过含1 mmol/L Na2S2O4的无糖Earle's溶液诱导神经元凋亡，并用MTT和Annexin V/PI双染法进行标记。此外，通过免疫荧光检测Bcl-2和Bax的蛋白表达。结果显示，对照组中观察到的凋亡细胞很少，而正常血浆组和低剂量SMA血浆组中凋亡细胞数量大幅增加。中剂量和高剂量SMA血浆均可保护培养的海马神经元免受OGD诱导的凋亡（P < 0.05）。高剂量SMA血浆的保护作用强于中剂量（P < 0.05）。与对照组、正常血浆组和低剂量SMA血浆组相比，中剂量和高剂量SMA血浆组的Bcl-2水平均显著升高（P < 0.05或0.01），而Bax的表达则相反。中剂量和高剂量SMA血浆组之间Bcl-2和Bax的表达无显著差异。Bcl-2和Bax阳性细胞数量有相似趋势。高剂量SMA血浆组的Bcl-2/Bax（阳性细胞数量）比值高于其他所有组（P < 0.05或0.01）。这些结果表明，含中高剂量SMA的血浆进行药理预处理可通过上调Bcl-2表达和下调Bax表达来提高OGD损伤海马神经元的活力并抑制其凋亡。
