乳腺癌的转移是其临床治疗面临的重点和难点,其侵袭性的高低受周围基质的影响很大.癌症相关成纤维细胞(CAFs)是肿瘤基质的重要成分,同时也是新型膜雌激素受体——G蛋白耦联雌激素受体(GPER)阳性细胞.研究表明,GPER可能是作用于癌细胞与肿瘤微环境(TM)的多种重要传导因子之间的关键交叉点,GPER在CAFs中活化对乳腺癌发生、发展和转移过程中发挥着重要的作用.多种中药单体的抗乳腺癌作用也与其靶向激活CAFs中的GPER通路是相关的.现就GPER在CAFs中的活化机制及其对乳腺癌转移的作用、中药单体对该过程的干预研究进行综述,以期为寻找乳腺癌治疗的新靶点、拓展乳腺癌症治疗策略提供思路.

This study explored the phytoestrogen-like effect of Siwu Decoction(SWD) and the estrogen receptor(ER)-mediated molecular mechanism based on network pharmacology and in vivo experiment. The active components and targets of SWD were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and related targets of "estrogen" from GeneCards and Online Mendelian Inheritance in Man(OMIM). Cytoscape and STRING were employed to construct the protein-protein interaction(PPI) network and "chemical component-target-disease" network and core targets were identified, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the core targets by R software. For the in vivo experiment, the 22-day-old SD female rats were treated(ig) with SWD for 4 days. Via hematoxylin-eosin(HE) staining, the morphological changes of rat uterus were observed. Reverse transcriptase-polymerase chain reaction(RT-PCR) was performed to detect mRNA expression of ER subtypes, estrogen-related targets, and the main regulatory factors in the estrogen signaling pathway. The results indicated 74 targets of SWD exerted phytoestrogen-like effect. KEGG pathway enrichment result suggested that estrogen signaling pathway was closely related to the phytoestrogen-like effect of SWD. Rats in SWD group demonstrated significantly thickened endometrium and significantly decreased expression of ERα, ERβ, and G protein-coupled estrogen receptor(GPER) mRNA in ovarian tissue. In addition, significant lowering of ERα and ERβ mRNA expression and significant rise of GPER mRNA expression in uterine tissue were observed in the SWD group. The expression of mitogen-activated protein kinase(MAPK) p38, MEK1/2 and extracellular signal-regulated kinase(ERK)1/2 mRNA was significantly low while that of epidermal growth factor receptor(EGFR) mRNA was significantly high in both ovarian and uterine tissues of SWD group compared with those in the control group. In conclusion, the phytoestrogen-like effect of SWD is closely related to the estrogen signaling pathway. The result lays a basis for revealing molecular mechanism of SWD in the treatment of gynecological diseases.

目的 探究人参皂苷Rg1对顺铂损伤大鼠卵巢颗粒细胞的保护作用及分子机制.方法 选取22～24 d SD的大鼠提取卵巢颗粒细胞进行原代培养,应用顺铂诱导建立卵巢早衰模型.设置正常组,模型组,人参皂苷Rg1低浓度组、中浓度组、高浓度组和雌二醇(E2)组.HE染色及免疫细胞化学法进行颗粒细胞鉴定;CCK8法检测人参皂苷Rg1作用24 h、48 h对顺铂损伤颗粒细胞的保护作用;Hoechst 33258染色检测细胞凋亡;Western blot法检测FSHR、PI3K、p-AKT、AKT、Bcl-2和Bax的蛋白表达.结果 细胞形态观察和胞质FSHR表达检测结果均表明所提取细胞为卵巢颗粒细胞;顺铂处理12h后,颗粒细胞增殖率呈剂量依赖性下降,而给予人参皂苷Rg1后,细胞增殖率下降被显著抑制,且呈剂量和时间依赖性;与模型组比较,人参皂苷Rg1和E2处理后卵巢颗粒细胞中FSHR、PI3K、p-AKT/AKT及Bcl-2/Bax蛋白表达水平显著上升,而PI3K抑制剂干预后,Bcl-2蛋白表达显著下降.结论 人参皂苷Rg1对顺铂损伤的卵巢颗粒细胞具有保护作用,该保护作用可能通过激活FSHR/PI3K/AKT通路,抑制卵巢颗粒细胞凋亡实现.

Abstract Background Bone protective effect of Si-Wu-Tang (SWT), a classical prescription of traditional Chinese medicine, is verified in clinical for thousand years. However, its mechanisms were still unclear. This study aims to investigate the molecular mechanism in ApoE -/- mice fed a high-fat diet combining network pharmacology and in-vivo experiments. Methods Femurs were collected from 6 ~ 8-week-old female ApoE-/- C57BL/6J mice (n = 12, 18–22 g) and their age-matched wild-type (WT) littermates C57BL/6J mice (n = 6, 18–20 g). They were divided into 3 groups: the control, SWT and model groups. Serum levels of high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by the serum biochemical index. HE staining and immunohistochemistry analysis were performed to observe the pathological tissue structure and the location and expression level of targets from the pathway screened out by the network pharmacology method. Western blot (WB) and RT-PCR analyses were performed to detect the expression levels of target proteins and mRNAs, respectively. Results The results of the network pharmacology analysis showed that the mechanism of SWT in treating osteopenia was closely related to the oestrogen receptor (ER) signalling pathway. In vivo experiments indicated that, compared with control group, the distribution of bone trabeculae was sparse, and the bone density decreased. The levels of HDL-C and LDL- C in the serum of the model group increased significantly (p < 0.01). The expression of GPER, PI3K, AKT and BCL-2 in the bone tissue of the model group decreased, and P53, BAX, ERα and ERβ were upregulated. Compared with the model group, the body mass of the SWT group increased slowly. The bone density and the distributions of bone trabeculae both increased. The expression of ERα, ERβ, GPER, PI3K, AKT and BCL-2 increased. The decreased expression of apoptotic genes, including P53 and BAX, was observed. Conclusion SWT significantly reduced bone loss in ApoE -/- mice fed a high-fat diet. An important mechanism might be that SWT could activate the PI3K/AKT signalling pathway mediated by ER and then inhibit apoptosis-related proteins to exert bone protective effects.

卵巢纤维化是多种妇科疾病的共同病理表现之一,卵巢早衰、子宫内膜异位囊肿、多囊卵巢综合征等均会造成卵巢纤维化;主要形成原因是卵巢组织细胞受损超过了周围实质细胞的修复能力,为了维持组织结构的完整性,细胞外基质大量产生并过度沉积,导致卵巢组织内结缔组织增多,实质细胞减少,最终造成卵巢不可逆修复而发生纤维化.卵巢纤维化的形成与许多细胞因子有关,本文将就卵巢纤维化相关疾病的影响因子、临床表现及其治疗方法的研究进展作简要介绍.

四物汤的应用从古至今已有悠久的历史,原为外伤"重伤肠内有淤血者"而设,后世多用于治疗血虚血滞等疾病.对于这一补血调经的良方,国内外学者进行了广泛而深入的研究,发现四物汤及组方中药的有效成分具有补血、调经、雌激素样作用、抗氧化、改善脑损伤、抗癌等多种药理作用,并且在妇科、骨科等临床方面的疗效较为显著.本文对近年来的研究成果进行了梳理和总结,将四物汤的药理作用和临床应用进行综述,以期为四物汤的深入研究提供理论依据,为临床用药提供参考.

目的 探讨四物汤在大鼠脑和骨骼肌组织中发挥雌激素样效应的机制.方法 将20只未达性成熟雌性SD大鼠完全随机分为正常对照组、雌激素组、四物汤高剂量组和低剂量组,各5只,适应性喂养4d后,正常对照组予相等剂量0.9％氯化钠溶液灌胃,雌激素组予戊酸雌二醇0.104 mg/(kg·d)灌胃,四物汤高剂量组予四物汤2.08 g/(kg·d)灌胃,四物汤低剂量组予四物汤0.52g/(kg·d)灌胃.每日早晚各灌胃1次,连续4d.各组大鼠最后一次灌胃后3h称重,10％水合氯醛腹腔麻醉后腹主动脉快速取血处死.观察大鼠脑及腿部骨骼肌组织细胞形态,采用蛋白质印迹法检测雌激素受体β(ERβ)、G蛋白偶联雌激素受体(GPER)、磷脂酰肌醇-3-激酶(PI3K)及蛋白激酶B(Akt)表达情况.结果 4组大鼠脑和骨骼肌组织细胞形态均未出现明显变化.在脑组织中,四物汤低剂量组ERβ表达量显著低于正常对照组和雌激素组[(0.354±0.066)比(0.450±0.022)、(0.509±0.042)];四物汤高剂量组和四物汤低剂量组GPER表达量显著低于雌激素组;四物汤低剂量组PI3K表达量显著低于正常对照组[(0.255±0.086)比(0.404 ±0.100)];雌激素组和四物汤高剂量组Akt表达量显著低于正常对照组[(0.224±0.049)、(0.268±0.097)比(0.467 ±0.102)](均P＜0.05或P＜0.01).在骨骼肌组织中,雌激素组和四物汤低剂量组ERβ表达量显著低于正常对照组;四物汤低剂量组PI3K表达量显著高于正常对照组和雌激素组,Akt表达量显著低于正常对照组和雌激素组(均P＜0.05或P＜0.01).结论 四物汤可不同程度抑制脑组织中ERβ、GPER、PI3K、Akt及骨骼肌中ERβ、Akt的表达,促进骨骼肌中PI3K的表达.推断四物汤可能通过ERβ和GPER介导的PI3 K/Akt信号通路发挥雌激素样效应.

目的:建立HPLC-ELSD指纹图谱研究牡蛎配伍生柴胡和醋柴胡后对柴胡皂苷类成分溶出的影响.方法:色谱条件:Phenomenex Kinetex C18色谱柱(250 mm×4.6 mm,5μm);流动相乙腈-水,梯度洗脱;流速1.0 ml · min-1;柱温30℃;进样量20μl;蒸发光散射检测器,氮气流速3.0 ml·min-1.分别对生柴胡单煎、生柴胡∶牡蛎(8∶3)、生柴胡∶牡蛎(1∶1)、醋柴胡单煎、醋柴胡∶牡蛎(8∶3)、醋柴胡∶牡蛎(1∶1)等6种水提液进行指纹图谱研究,得到共有模式色谱图后进行比对,并测定柴胡皂苷a,d,b1,b2的含量.结果:柴胡配伍牡蛎后,新生成1个化学成分(10号峰),并且大幅度升高柴胡皂苷d的煎出率,同时显著降低柴胡皂苷a(SSa)、柴胡皂苷b1(SSb1)、柴胡皂苷b2(SSb2)、柴胡皂苷d(SSd)等7种成分的煎出率(16～ 22号色谱峰),并且牡蛎配伍比例越大,这7个成分减少得越多甚至消失.结论:本研究建立的HPLC-ELSD指纹图谱能够较为全面地分析柴胡配伍牡蛎后柴胡中主要成分的变化,揭示柴胡与牡蛎的配伍规律.