丹参酮ⅡA 预防慢性缺氧大鼠认知功能障碍的电生理机制

目的:探讨丹参酮ⅡA(TⅡA)预防慢性缺氧大鼠认知功能障碍的电生理机制。方法:将18只雄性SD大鼠(200-250 g)随机分为对照组、模型组(Model组)、TⅡA(10mg/kg.d)治疗组(TⅡA组)。复制慢性缺氧大鼠认知功能障碍模型,并给予相应治疗,在脑片水平运用膜片钳技术检测海马CA1区的LTP变化,并检测海马CA1区锥体细胞的兴奋性变化。结果:(1)给予高频强直刺激(HFS)后各组兴奋性突出后电位(fEPSP)斜率均显著增加,即均可诱发LTP并持续1h以上,但模型组LTP较对照组显著减弱(P<0.05),TⅡA治疗组LTP较模型组明显增强(P<0.05);(2)慢性缺氧使海马CA1锥体细胞放电所需的刺激电流幅度显著增加、阈电位升高、兴奋性降低,同样刺激强度条件下动作电位数量减少,TⅡA干预可明显减轻慢性缺氧对海马CA1锥体细胞的上述抑制。结论:TⅡA可能是通过维持海马CA1锥体细胞的兴奋性、维持海马的突出可塑性减轻慢性缺氧对认知功能的损害。     

The anti-atherosclerotic effect of tanshinone IIA is associated with the inhibition of TNF-α-induced VCAM-1, ICAM-1 and CX3CL1 expression

Tanshinone IIA is one of the major diterpenes in Salvia miltiorrhiza. The inhibitory effect of Tanshinone IIA on atherosclerosis has been reported, but the underlying mechanism is not fully understood. The present study aimed to study the anti-atherosclerosis effect of Tanshinone IIA on the adhesion of monocytes to vascular endothelial cells and related mechanism. Results showed that Tanshinone IIA, at the concentrations without cytotoxic effect, dose-dependently inhibited the adhesion of THP-1 monocytes to the TNF-α-stimulated human vascular endothelial cells. The expressions of cell adhesion molecules including VCAM-1, ICAM-1 and E-selectin were induced by TNF-α in HUVECs at both the mRNA and protein levels. The mRNA and protein expressions of VCAM-1 and ICAM-1, but not E-selectin, were both significantly suppressed by Tanshinone IIA in a dose dependent manner. In addition, the TNF-α-induced mRNA expression of fractalkine/CX3CL1 and the level of soluble fractalkine were both reduced by Tanshinone IIA. We also found that Tanshinone IIA significantly inhibited TNF-α-induced nuclear translocation of NF-κB which was resulted from the inhibitory effect of Tanshinone IIA on the TNF-α-activated phosphorylation of IKKα, IKKβ, IκB and NF-κB. As one of the major components of Salvia miltiorrhiza, Tanshinone IIA alone exerted more potent effect on inhibiting the adhesion of monocytes to vascular endothelial cells when compared with Salvia miltiorrhiza. All together, these results demonstrate a novel underlying mechanism for the anti-inflammatory effect of Tanshinone IIA by modulating TNF-α-induced expression of VCAM-1, ICAM-1 and fractalkine through inhibition of TNF-α-induced activation of IKK/NF-κB signaling pathway in human vascular endothelial cells.     

Biological evaluation of tanshindiols as EZH2 histone methyltransferase inhibitors

EZH2 is the core subunit of Polycomb repressive complex 2 catalyzing the methylation of histone H3 lysine-27 and closely involved in tumorigenesis. To discover small molecule inhibitors for EZH2 methyltransferase activity, we performed an inhibitor screen with catalytically active EZH2 protein complex and identified tanshindiols as EZH2 inhibitors. Tanshindiol B and C potently inhibited the methyltransferase activity in in vitro enzymatic assay with IC₅₀ values of 0.52 μM and 0.55 μM, respectively. Tanshindiol C exhibited growth inhibition of several cancer cells including Pfeiffer cell line, a diffuse large B cell lymphoma harboring EZH2 A677G activating mutation. Tanshindiol treatment in Pfeiffer cells significantly decreased the tri-methylated form of histone H3 lysine-27, a substrate of EZH2, as revealed by Western blot analysis and histone methylation ELISA. Based on enzyme kinetics and docking studies, we propose that tanshindiol-mediated inhibition of EZH2 activity is competitive for the substrate S-adenosylmethionine. Taken together, our findings strongly suggest that tanshindiols possess a unique anti-cancer activity whose mechanism involves the inhibition of EZH2 activity and would provide chemically valuable information for designing a new class of potent EZH2 inhibitors.     

丹参酮IIA缓解大鼠心肌梗死后左心室重构的作用及其机制研究

摘要 目的：探讨丹参酮IIA（T-IIA）对于缓解大鼠心肌梗死（MI）后左心室重构（LVR）的作用及其机制。方法：选取SD雄性大鼠80只,通过结扎左前降支（LAD）建立MI大鼠模型。将大鼠随机分为8组,假手术组未结扎LAD,其余各组均结扎LAD;除假手术组和MI组腹腔注射生理盐水外,其余各组分别给予T-IIA、脂多糖（LPS）和TAK-242治疗。HE和马松（Masson）三色染色评估MI大小、组织病理改变和纤维化程度。末端dUTP镍末端标记（TUNEL）染色观察心肌细胞凋亡情况。采用反转录定量聚合酶链反应(RT-qPCR)和蛋白免疫印迹试验检测Toll样受体4（TLR4）、髓样分化蛋白88（MyD88）和核因子κB（NF-κB）的表达水平。结果：T-IIA能改善MI大鼠心功能,可降低MI大鼠心脏体积,改善心脏形态,减轻MI大鼠的组织病理学改变,并有效减轻MI和心肌纤维化。T-IIA抑制MI大鼠的TLR4/MyD88/NF-κB信号通路,且能有效减少MI大鼠梗死边缘区心肌细胞凋亡。结论：T-IIA通过抑制TLR4/MyD88/NF-κB信号通路的激活,改善心脏形态、功能和病理组织学变化,有效减轻MI的严重程度,预防LVR。     

丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的实验研究

目的:探究丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的机制。方法:体外培养人肝癌细胞Hep3B、HepG2,并分为:实验组、阳性对照组和空白对照组,实验组使用丹参酮处理,阳性对照组使用阿霉素处理,空白对照为加入10μL DMSO或生理盐水,使用CCK8检测肝癌细胞的增殖情况;分别加入浓度为31μmol/L和2.5μmol/L的丹参酮及阿霉素显微镜下观察细胞形态变化;使用流式细胞术检测肝癌细胞的凋亡情况和细胞周期情况;使用Western blot检测肝癌细胞VEGF及VEGFR蛋白表达情况。结果:MTT实验结果显示,随着丹参酮和阿霉素使用浓度的升高人肝癌细胞Hep3B、HepG2生长受到显著的抑制(P0.05);显微镜下观察发现丹参酮可以抑制肝癌肿瘤细胞增殖;流式细胞检测发现相比空白对照组,阳性对照组和实验组(人肝癌细胞Hep3B、HepG2)细胞凋亡率显著增加(P0.05);相比空白对照组,实验组和阳性对照组G0/G1期细胞百分比显著增加(P0.05),S期和G2/M期细胞百分比显著降低(P0.05);蛋白质印记实验结果显示,相比空白对照组,实验组和阳性对照组VEGF及VEGFR蛋白表达显著降低(P0.05),且实验组表达显著低于阳性对照组(P0.05)。结论:参酮通可以通过抑制VEGF/VEGFR信号通路,将肝癌细胞分裂阻滞在G0/G1,达到抑制肝癌细胞的增殖及迁移和侵袭能力的效果。     

Tanshinone IIA, an isolated compound from Salvia miltiorrhiza Bunge, induces apoptosis in HeLa cells through mitotic arrest

AIMS: Tanshinone IIA (Tan IIA) is a compound isolated from Salvia miltiorrhiza Bunge (Danshen). The aim of this study is to investigate the mechanisms of its anti-cancer effect. MAIN METHODS: To clearly delineate the cell cycle-dependent effects of Tan IIA, we used either synchronized cells or single living cell analysis to conduct our studies. Subcellular fractionation, Western blot analysis, immuno-fluorescence staining and FACS analysis were also employed in our study. KEY FINDINGS: We found that Tan IIA could arrest cancer cells in mitosis by disrupting the mitotic spindle and subsequently triggered cells to enter apoptosis through the mitochondria-dependent apoptotic pathway. Thus, Tan IIA could selectively kill mitotic cells over interphase cells. In comparison with other existing anti-cancer drugs that cause mitotic arrest by interfering with the microtubule structure (such as vincristine or taxol), Tan IIA destroyed only the mitotic spindle during the M phase but not the microtubule structure in interphase cells. Furthermore, Tan IIA could trigger the mitotic arrested cells to enter apoptosis faster than vincristine or taxol. SIGNIFICANCE: Since Tan IIA can selectively induce cancer cells to enter apoptosis through mitotic arrest, it has the potential to be developed into an anti-cancer drug.     

Tanshinone IIA alleviates lipopolysaccharide‐induced acute lung injury by downregulating TRPM7 and pro‐inflammatory factors

The study aimed to investigate the role of Tanshinone IIA (Tan IIA) in lipopolysaccharide (LPS)‐induced acute lung injury (ALI) in its regulation of TRPM7. Wistar male rats were randomly divided into the normal saline (NS), LPS, knockout (KO) + LPS, low‐dose Tan IIA (Tan‐L), middle‐dose Tan IIA (Tan‐M), high‐dose Tan IIA (Tan‐H) and KO + high‐dose Tan IIA (KO + Tan‐H) groups. The level of tumour necrosis factor‐α (TNF‐α), interleukin (IL)‐1β, IL‐6, TRPM7 protein expression, current density‐voltage curve and Ca²⁺ concentration were detected through ELISA, Western blotting, electrophysiological experiment and a calcium‐imaging technique, respectively. The rats in the KO + LPS, Tan‐L, Tan‐M, Tan‐H and KO + Tan‐H groups all displayed lower levels of TNF‐α, IL‐1β and IL‐6 than the LPS group. Rats in the KO + Tan‐H group exhibited lower levels of NF‐α, IL‐1β and IL‐6 than rats in the Tan‐H group. Elevated levels of TRPM7 protein expression in the LPS and Tan groups were detected in comparison with the NS group. However, TRPM7 protein expression in Tan‐M and Tan‐H groups was notably lower than in that of the LPS group. In comparison with the NS group, the LPS and Tan groups had a greater PIMs cell density and a higher concentration of Ca²⁺. Contrary results were observed in the KO + LPS, Tan‐H and KO + Tan‐H groups. Tan IIA decreases calcium influx in PIMs and inhibits pro‐inflammatory factors which provide an alleviatory effect in regards to LPS‐induced ALI by suppressing TRPM7 expression.     

美沙拉嗪联合丹参酮胶囊对活动期溃疡性结肠炎疗效分析

目的：探讨美沙拉嗪联合丹参酮胶囊对活动期溃疡性结肠炎患者血浆D-二聚体、CRP、凝血功能的影响及临床疗效。方法： 将90 例活动期溃疡性结肠炎患者按照随机数字表随机分为Ⅰ组、Ⅱ组及Ⅲ组，每组30 例，Ⅰ组给予口服柳氮磺胺吡啶肠溶片治 疗，Ⅱ组给予口服美沙拉嗪肠溶片治疗，Ⅲ组给予口服美沙拉嗪肠溶片+ 丹参酮胶囊治疗，三组均治疗4 周；检测各组患者治疗 前后凝血功能指标如活化部分凝血酶原时间(APTT)、纤维蛋白原(FIB)、血小板计数(PLT)、血小板平均体积(MPV)、血小板最大聚 集率[PAG(Max)]和D -二聚体(D-D)、C- 反应蛋白(CRP)，Mayo 疾病活动指数评分；比较三组之间的差异。结果：与Ⅰ及Ⅱ组比 较，Ⅲ组PLT、PAG(Max)及D-D 在治疗2、4 周后明显降低(P＜0.05)，APTT 及MPV 在治疗4 周后升高(P＜0.05)；FIB 及CRP 亦 在治疗4 周后降低(P＜0.05)；三组治疗后Mayo 指数评分均降低，但Ⅲ组治疗4 周后显著低于Ⅰ及Ⅱ组(P＜0.05)，其中Ⅲ组治疗 后Mayo 指数评分降低幅度(△Mayo 指数评分)大于Ⅰ及Ⅱ组(P＜0.05)。结论：美沙拉嗪联合丹参酮胶囊能够显著改善活动性溃 疡性结肠炎患者的炎症及高凝状态，在改善患者的临床症状方面优于单一用药。     

Tanshinone IIA increases recruitment of bone marrow mesenchymal stem cells to infarct region via up-regulating stromal cell-derived factor-1/CXC chemokine receptor 4 axis in a myocardial ischemia model

Systemic administration with bone marrow mesenchymal stem cells (BMSCs) is a promising approach to cure myocardial ischemia (MI), while the efficacy of cell transplantation is limited by the low engraftment of BMSCs. Tanshinone IIA (Tan IIA) has been reported many times for the treatment of MI. Therefore, the present study was performed to investigate whether Tan IIA could increase the migration of BMSCs to ischemic region and its potential mechanisms. In our study, we found that combination treatment with Tan IIA and BMSCs significantly alleviated the infarct size when compared with control group (31.46 ± 3.00% vs. 46.95 ± 6.51%, p < 0.05). Results of real-time PCR showed that Tanshinone IIA (Tan IIA) did increase the migration of BMSCs to ischemic region in vivo, which was correlated with cardiac function recovery after MI. Furthermore, 2 μM Tan IIA could enhance the migration capability of BMSCs in vitro (3.69-fold of control), and this enhancement could be blocked by AMD3100 (a CXC chemokine receptor 4 blocker). CXCR4, together with its specific receptor, stromal cell-derived factor-1 (SDF-1) plays a critical role in the stem cell recruitment. Our experiment indicated that Tan IIA could promote SDF-1α expression in the infarct area and enhance the CXCR4 expression of BMSCs in vitro. Therefore, we postulated that Tan IIA could increase the BMSCs migration via up-regulating SDF1/CXCR4 axis.     

丹参酮IIA对大鼠移植性肝癌的抑制作用

目的：观察丹参酮IIA对大鼠移植性肝癌的抑制作用。方法：将sD大鼠随机分为假手术组、模型组、丹参酮[IA高、低剂量组。各组动物于移植术后12d处死。分离肿瘤测量其体积,reahimeRT．PCR法和western blot法检测肿瘤组织中HMGB1、VEGFmRNA和蛋白的表达。结果：丹参酮IIA能明显降低肿瘤体积,降低肿瘤组织中HMGBl、VEGFmRNA和蛋白的表达。结论：丹参酮IIA能抑制大鼠移植性肝癌的生长,其机制可能与抑制HMGB1、VEGF表达有关。