Background: Endoplasmic reticulum stress (ERS) is a key part of the apoptotic cascade that is initiated after cerebral ischemia-reperfusion injury and is very important for research on poststroke rehabilitation. In addition, the unfolded protein response (UPR) plays an important role in ERS because it activates downstream apoptotic signal transduction and induces apoptosis through the glucose-regulated protein 78 (GRP78)/protein kinase R (PKR)-like ER kinase (PERK)/activating transcription factor 4 (ATF4) pathway. The Gua Lou Gui Zhi Decoction (GLGZD) ameliorated neuronal apoptosis of ischemia-reperfusion injury caused by middle cerebral artery occlusion (MCAO) had been proved in our previous study. The present study aims to underly the regulatory ability of GLGZD in ERS-induced apoptosis mediated by the GRP78/PERK/ATF4 pathway. Methods: GLGZD was analyzed by HPLC. The effects of GLGZD were obversed on MCAO-induced ischemic rats. The cerebral infarct volume was detected by 2,3,5-Triphenyl-2H-Tetrazolium Chloride (TTC) Staining. Terminal Deoxynucleotidyl Transferase-Mediated dUTP-Biotin Nick End Labeling (TUNEL) were used to detect apoptosis. Transmission Electron Microscopy (TEM), Ca2+ levels and reactive oxygen species (ROS) detection were used to determine the function of endoplasmic reticulum. The GRP78/PERK/ATF4 signaling pathway was assessed by western blotting and immunohistochemistry. Results: Our results showed that GLGZD exerted its effects on ischemia-reperfusion injury by significantly promoting the restoration of the quantity and morphology of the rough ER and reducing the neuronal apoptosis rate in the ischemic cortex. Moreover, both of the intracellular ROS and Ca2+ levels in ischemic cortical cells were found significantly reduced by GLGZD. The GLGZD-treated group showed increased levels of phosphorylation in both of PERK and eukaryotic translation initiation factor 2α (eIF2α), activation of cysteinyl aspartate-specific proteinase-3 (Caspase-3), upregulation of the total protein levels of sarcoplasmic/endoplasmic Ca2+ ATPase 2α (SERCA 2α) and B-cell leukemia/lymphoma gene 2 (Bcl-2). Conclusions: These findings suggest that GLGZD reduces oxidative stress-induced injury and promotes a dynamic calcium balance, thereby inhibiting ERS and exerting an antiapoptotic effect on neuronal ischemic injury, which are closely related to the activation of GRP78/PERK/ATF4 signaling pathway.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的观察蒲郁胶囊对雄性大鼠性功能与血清雄激素的影响,并与米氮平和药氟西汀进行比较,研究蒲郁胶囊的药理学特点。方法 60只雄性大鼠随机分为空白、氟西汀、米氮平和蒲郁胶囊低、中、高剂量共6组。重复给药1周和2周后,通过大鼠交配行为模型和非接触行为模型观察大鼠性功能的差异;之后分别加入五羟色胺1A受体拮抗剂WAY100635和部分激动剂丁螺环酮观察交配行为和非接触性阴茎勃起行为;最后取血,分离血清,ELISA法检测各组雄性大鼠血清雄激素水平。结果氟西汀对性功能有抑制作用,而蒲郁胶囊和米氮平对大鼠性功能均无影响;3种药物对大鼠雄激素均无影响。加入WAY100635和丁螺环酮后能逆转氟西汀引起的性功能障碍,对蒲郁胶囊无影响。结论蒲郁胶囊对雄性大鼠雄激素与性功能的影响均较小,优于氟西汀等SSRIs类抗抑郁药物,其原因可能与其不影响5-HT有关。</span>

Cerebral ischemia–reperfusion injury is a complex pathophysiological process. Poly(ADP-ribose) (PAR) polymerase-1 (PARP-1)/apoptosis-inducing factor (AIF) signaling pathway-mediated apoptosis is one of the non-caspase-dependent cell death programs that are widely present in neurological diseases such as stroke. In our study, we aimed to conduct further research on the effects of Gualou Guizhi decoction (GLGZD) on the PARP-1/AIF signaling pathway in cell apoptosis after ischemia–reperfusion injury caused by middle cerebral artery occlusion (MCAO). The results showed that GLGZD administration for 7 days significantly ameliorated MCAO-induced neurological damage, limb paralysis and the pathological state of the ischemic cortex. GLGZD exerted its effects by significantly reducing the volume of ischemic cerebral infarction, increasing the number of Nissl-positive cells, and reducing neuronal apoptosis. Furthermore, Western blot analysis showed that GLGZD significantly inhibited the total protein expression of PARP-1, PAR, AIF and endonuclease G (Endo G) in the ischemic cortex and significantly increased the total protein expression of heat-shock protein 70 (Hsp70). On the one hand, the expression of PARP-1, AIF and Endo G protein in the nucleus significantly decreased while the expression of PAR nucleoprotein significantly upregulated. On the other hand, compared with the MCAO model group, the GLGZD-treated group showed a significantly reduced protein expression of PAR in mitochondria and significantly increased protein expression of mitochondrial AIF and Endo G. It was concluded that GLGZD had good therapeutic effects in MCAO model rats. These effects were closely related to GLGZD-mediated inhibition of ischemia-induced neuronal apoptosis by regulation of protein expression and translocation in the PARP-1/AIF signaling pathway.

目的 探讨栝楼桂枝汤对脑缺血再灌注损伤模型大鼠皮质内质网应激相关因子rmRNA表达的影响. 方法 将大鼠随机分为假手术组和造模组,造模组采用线栓法制备大鼠大脑中动脉阻塞(MCAO)模型,假手术组除不插入线栓外其余操作相同.将造模成功的大鼠随机分为模型组、阳性对照组、低剂量组、中剂量组和高剂量组.低、中、高剂量组分别予不同剂量的栝楼桂枝汤灌胃,阳性对照组给予尼莫地平混悬液灌胃,假手术组和模型组予等量生理盐水灌胃,每日1次,连续干预7d.采用Longa神经学评分法评定大鼠神经功能损伤程度,RT-qPCR法检测大鼠缺血侧皮质组织中葡萄糖调节蛋白78(GRP78)、转录活化因子4(ATF4)、C/EBP同源蛋白(CHOP)及其B淋巴细胞瘤-2基因(Bcl-2)、半胱氨酸天冬氨酸蛋白-3(Caspase-3)mRNA的表达. 结果 与假手术组比较,模型组神经功能损伤评分及大鼠皮质组织中GRP78、ATF4、CHOP、Caspase-3 mRNA表达明显提高(P＜0.01,P＜0.05),Bcl-2 mRNA表达降低(P＜0.05);与模型组比较,阳性对照组和低、中、高剂量组神经功能损伤评分及大鼠皮质组织中GRP78、ATF4、CHOP、Caspase-3 mRNA表达明显下降(P＜0.01,P＜0.05),Bcl-2mRNA表达提高(P＜0.05). 结论 栝楼桂枝汤对MCAO模型大鼠具有较好的治疗作用,其作用机制可能与调控内质网应激相关因子的表达,降低内质网应激水平有关.

目的 从解郁合欢汤、血府逐瘀汤和天王补心丹3种复方水煎剂中初步筛选出有潜在抗抑郁效果的复方. 方法 取健康SPF级雄性KM小鼠50只,根据旷场试验结果随机分成空白组、盐酸氟西汀组、解郁合欢汤组、血府逐瘀汤组和天王补心丹组5组,每日灌胃给药1次,连续给药14d.每周进行悬尾试验、强迫游泳试验观测小鼠行为学变化;给药结束后测定小鼠自发活动;行为学试验结束后,用Western Blot法测定小鼠海马p-GSK3β的表达量. 结果 ①与空白组相比,血府逐瘀汤组和天王补心丹组在给药第1周和第2周均能显著缩短小鼠悬尾和游泳不动时间,作用与盐酸氟西汀组相当,其中天王补心丹组作用最显著;解郁合欢汤组有作用趋势,但无显著性差异.②3种复方均不影响小鼠自发活动.③3种复方均能增加小鼠海马p-GSK3β的表达量,天王补心丹组作用最显著. 结论 天王补心丹抗抑郁效果较强,其作用机制可能与增加海马p-GSK3β的表达有关.

目的 观察栝楼桂枝汤对大鼠脑缺血再灌注损伤后脑组织中多聚(腺苷二磷酸核糖)聚合酶-1(PARP-1)表达的影响,探讨栝楼桂枝汤的神经保护机制. 方法 用线栓法制备大脑中动脉闭塞大鼠模型,手术当日大鼠清醒后用栝楼桂枝汤干预,每日1次,7天后用Longa评定法评价神经功能损伤程度,TTC染色检测脑梗死体积,HE染色检测缺血侧大脑皮层的病理改变,免疫组化法检测缺血侧大脑皮层PARP-1的表达. 结果 与模型组比较,栝楼桂枝汤组能明显改善MCAO模型大鼠的神经功能缺损症状,缩小脑梗死体积,减轻大脑皮层病理损伤,减少大脑皮层PAPR-1的表达. 结论 栝楼桂枝汤能有效降低缺血再灌注后脑组织的PARP-1表达,具有较好的神经保护作用.

Gualou Guizhi decoction (GLGZD) is effective for the clinical treatment of limb spasms caused by ischemic stroke, but its underlying mechanism is unclear. Propidium iodide (PI) fluorescence staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemistry, western blot, and real-time qPCR were used to observe the axonal regeneration and neuroprotective effects of GLGZD aqueous extract on organotypic cortical slices exposed to oxygen-glucose deprivation (OGD) and further elucidate the potential mechanisms. Compared with the OGD group, the GLGZD aqueous extract decreased the red PI fluorescence intensity; inhibited neuronal apoptosis; improved the growth of slice axons; upregulated the protein expression of tau and growth-associated protein-43; and decreased protein and mRNA expression of neurite outgrowth inhibitor protein-A (Nogo-A), Nogo receptor 1 (NgR1), ras homolog gene family A (RhoA), rho-associated coiled-coil-containing protein kinase (ROCK), and phosphorylation of collapsin response mediator protein 2 (CRMP2). Our study found that GLGZD had a strong neuroprotective effect on brain slices after OGD injury. GLGZD plays a vital role in promoting axonal remodeling and functional remodeling, which may be related to regulation of the expression of Nogo-A and its receptor NgR1, near the injured axons, inhibition of the Rho-ROCK pathway, and reduction of CRMP2 phosphorylation.

Objective:To determine the content of Spinosin in Honey-fried Semen Zizyphi Spinosae(Mizhi Suanzaoren)by HPLC (High Performance Liquid Chromatography).Methods:The experiment was conducted in the condition of WondaCract ODS-2(250 mm ×4.6 mm,5μm),mobile phase isacetonitrile-water,detection wavelength 335nm,and flow velocity 1 mL/min,column tempera-ture 30 ℃,using RP-HPLC.Results:The linearity range of Spinosin is 0.005 ~0.3 mg/mL,r =0.9965,average recovery rate is 99.74%,RSD =1.46%.Conclusion:This method is accurate with great reproducibility.It can offer testing standards for process-ing study and quality control of Honey-fried Semen Zizyphi Spinosae.

目的 观察寒痹方离子导入液的皮肤刺激性和抗炎镇痛作用. 方法 采用家兔同体左右侧自身比较法研究皮肤刺激性;采用二甲苯致小鼠耳肿胀实验、热板致小鼠疼痛和醋酸致小鼠疼痛实验,比较寒痹方离子导入液经皮外用和直流电离子导入后的抗炎镇痛作用. 结果 寒痹方离子导入液对家兔的正常皮肤无刺激性,仅1只家兔的破损皮肤处出现轻度红斑,但在停药24 h后恢复正常;离子导入组较空白对照组和未导入组的耳肿胀度明显减轻、痛阈值明显提高、扭体次数明显减少(P＜0.05或0.01);离子导入组的痛阈提高百分率(97.57％)和镇痛百分率(56.02％)均高于未导入组(50.48％、33.77％). 结论 寒痹方离子导入液对家兔皮肤无明显刺激性,具有较好的抗炎镇痛作用,其作用优于未导入组.

Objectve To choose the optimal processing technology of honey-fired Semen Ziziphi Spinosae. Methods Through multiple indexes such as appearance, content of water extract, fatty oil, flavonoids, spinosin and jujuboside A, choose the best processing technology of honey-fired Semen Ziziphi Spinosae according to L9 (34) orthogonal experiment design including three factor such as temperature, time and the ratio of honey, moreover design four levels for each factor. Results Through process optimization, the best processing technology of honey-fired Semen Ziziphi Spinosae is heating decoction pieces on the temperature of 150 ℃ in 4.5 min adding honey by 7.5% ratio. Conclusion The optimal processing technology of honey-fired Semen Ziziphi Spinosae is stable and feasible.

[目的]建立蜜炙酸枣仁中酸枣仁皂苷A的含量测定方法.[方法]采用反相高效液相色谱法,以WondaCract ODS-2(250 mm×4.6 mm,5μm)为色谱柱,乙腈-水为流动相,ELSD漂移管温度为80℃,气体流量为1.5 ml/min.紫外检测波长为204nm;流速为1 ml/min,柱温为30℃.[结果]HPLC-ELSD法测定酸枣仁皂苷A的线性范围为0.05～0.2 mg/ml,r=0.9952,平均回收率为102.97％,RSD=0.79％(n=6).HPLC-UV法测定酸枣仁皂苷A的线性范围为0.005～0.2 mg/ml,r=0.9990,平均回收率为98.64％,RSD=2.05％(n=6).[结论]两种检测方法测定酸枣仁皂苷A的含量结果相近,但HPLC-ELSD法的稳定性更好,HPLC-UV法由于吸收度低,峰面积相对较小,存在一定误差.