In this study, we explored the pharmacological effects of Siwu Decoction in treating premature ovarian insufficiency(POI) and its molecular mechanism based on the mitophagy pathway modulated and mediated by estrogen receptor(ER) subtypes. Female Balb/c mice were divided into a control group, model group, as well as high-dose and low-dose groups of Siwu Decoction. The POI mice model was constructed by intraperitoneal injection of cisplatin. The high-dose and low-dose groups of Siwu Decoction were administered intragastrically with Siwu Decoction each day for 14 days. During this period, we monitored the estrous cycle and body weight of the mice and calculated the ovarian index. The morphology of the ovaries was detected by hematoxylin-eosin(HE) staining, and the number of primordial follicles was counted. The apoptosis of the ovarian tissue was detected by TUNEL staining. The expression levels of anti-Müllerian hormone(AMH), apoptosis-associated and mitophagy-associated proteins, ER subtypes, and the expression levels of key proteins of its mediated molecular pathways were detected by Western blot and immunohistochemistry. KGN cells were divided into a control group, model group, Siwu Decoction group, and gene silencing group. The apoptosis model was induced by H_2O_2, and PTEN-induced putative kinase 1(PINK1) gene silencing was induced by siRNA transfection. The Siwu Decoction group and gene silencing group were added to the medium containing Siwu Decoction. Cell viability was detected by CCK-8 assay. Cell senescence was detected by senescence-associated-β-galactosidase. The expression levels of apoptosis-associated and mitophagy-associated proteins were detected by Western blot. The results of in vivo experiments showed that compared with the model group, the mice in the high-dose and low-dose groups of Siwu Decoction significantly recovered the rhythm of the estrous cycle, and the levels of ovarian index, number of primordial follicles, and expression of AMH, representative indexes of ovarian function, were significantly higher, suggesting that the level of ovarian function was significantly improved. The expression levels of the apoptosis-related proteins, cytochrome C(Cyt C), cysteinyl aspartate specific proteinase 3(caspase 3), B-cell lymphoma-2(Bcl-2)-associated X(Bax), and mitophagy-associated indicator(Beclin 1) were significantly decreased, and the expression levels of Bcl-2 was significantly elevated. The positive area of TUNEL was significantly reduced, suggesting that the apoptosis level of the ovaries was significantly reduced. The expression levels of PINK1, Parkin, and sequestosome 1(p62) were significantly reduced, suggesting that the level of ovarian mitophagy was significantly down-regulated. The expression levels of ERα and ERβ were significantly elevated, and the ratio of ERα/ERβ was significantly reduced. The expression levels of key proteins in the pathway, phosphoinositide 3-kinase(PI3K) and protein kinase B(Akt), were significantly reduced, suggesting that the regulation of ER subtypes and the mediation of PI3K/Akt pathway were the key mechanisms. In vitro experiments showed that compared with the model group, the proportion of senescent cells in the Siwu Decoction group was significantly reduced. Cyt C, caspase 3, Beclin 1, Parkin, and p62 were significantly reduced, which was in line with in vivo experimental results. The proportion of senescent cells and the expression level of the above proteins were further significantly reduced after PINK1 silencing. It can be seen that Siwu Decoction can regulate the expression level and proportion of ER subtypes in KGN cells, then mediate the PI3K/Akt pathway to inhibit excessive mitophagy and apoptosis, and exert therapeutic effects of POI.

BackgroundFumigation and sitz-bath therapy are traditional Chinese medicine (TCM) practices that offer unique benefits for managing chronic perianal eczema (CPE). This study aimed to investigate the clinical efficacy of the traditional Chinese herbal lotion (anal pruritus lotion, APL) combined with pimecrolimus cream in CPE treatment.MethodsPatients with CPE admitted between October 2019 and March 2022 were analyzed. The control group was given basic therapy with pimecrolimus cream, whereas the treatment group received pimecrolimus cream combined with APL under fumigation and sitz-bath therapy. We recorded and compared baseline patient information and clinical symptoms pre- and post-therapy, including clinical symptom scores, the eczema area and severity index (EASI), pruritus visual analogue scale (VAS) scores, dermatology life quality index (DLQI), and efficacy evaluations. Additionally, safety assessments and follow-up surveys were performed, as well.ResultsBaseline data were comparable between the treatment and control groups. Post-therapy, the treatment group exhibited significantly improved outcomes compared to the control group in eczema severity, pruritus VAS scores, and dermatology life quality, with lower relapse rates (P < 0.05). Safety evaluations suggested that the treatments were safe and reliable.ConclusionThe combination of pimecrolimus cream and APL is more effective in the treatment of CPE than pimecrolimus cream alone, providing a promising new approach for the clinical management of perianal eczema.Clinical Trial registrationInternational Traditional Medicine Clinical Trial Registry, http://itmctr.ccebtcm.org.cn/en-US/Home/ProjectView?pid=58f1a168-bdf1-4e2a-a9bd-cf0f4a3ec06f as ITMCTR 2024000576.

Ethnopharmacological relevance: Siwu decoction (SWD) is widely used in gynecological diseases, such as peripheral menopause syndrome, premature ovarian failure, and menstrual disorder. However, the mechanism of SWD on postmenopausal osteoporosis (PMOP) remains unclear. Aim of the study: To discover the phytoestrogenic osteoprotective effect of SWD on PMOP. Materials and methods: The potential mechanism of SWD on PMOP was filtered through network pharmacology research. The potential mechanism was verified in MC3T3-E1 cell lines in vitro. CCK8 assay was conducted to assess cell proliferation and the expressions of ER/PI3K/AKT pathway were analyzed using Western blot. Female F-344 rats were chosen to set up the PMOP model. The osteoprotective effect of SWD in vivo was evaluated using Hematoxylin-eosin staining, TRAP staining, Goldner staining and DXA. The potential mechanism was verified in vivo through Western blot and immunohistochemistry. RT-qPCR was conducted to unveil the expressions of osteogenesis genes. Results: Network pharmacology research showed that ER/PI3K/AKT pathway may be the potential mechanism of SWD on PMOP. SWD promoted the proliferation of osteoblasts and regulated the protein expressions of ER/PI3K/ AKT pathway in vitro. SWD improved the morphological structure, bone mineralization and bone mineral density of femurs and suppressed osteoclastogenesis in PMOP rat model via ER/PI3K/AKT pathway in vivo. In addition, SWD regulated the mRNA expressions of osteogenesis-related genes. Conclusions: SWD exerts a phytoestrogenic osteoprotective on PMOP by regulating ER/PI3K/AKT pathway, which marks it as a valuable medicine or supplement of PMOP.

The targets and mechanisms of Si-Wu-Tang (SWT) against (Breast cancer) BRCA were identified and a survival model and nomogram was construted by network pharmacology, bioinformatic analysis and in vitro experiments. A total of 72 anti-breast cancer SWT targets were selected, among which eleven genes (MAOA、SQLE、CACNA2D1、GLI1、RORB、ITGB3、TACR1、NR3C2、CA3、RBP4 and PTK6) were used to construct a novel prognostic model of breast cancer. The anti-breast cancer activity of SWT was related to the modulation of the receptor tyrosine kinases signaling pathways. Moreover, two compounds, mairin and senkyunone were found to bind directly to ITGB3 and RORB proteins. Finally, mRNA and protein expression of ITGB3 and RORB was observed to be significantly down-regulated after incubation of MCF-7 cells with SWT. Overall, our results indicated that mairin and senkyunone were the key ingredients present in SWT, and ITGB3 as well as RORB proteins were the major targets affected by SWT. The prognostic model can be used to predict the outcome of BRCA patients.

Due to the lack of classic estrogen receptors, there has been a shortage of targeted therapy for triple-negative breast cancer (TNBC), resulting in a poor prognosis. However, the newly discovered G protein-coupled estrogen receptor (GPER) has been found to be expressed in TNBC cells. Salvia miltiorrhiza (Danshen) is an essential Chinese medicine for gynecological disorders, and its component tanshinone IIA (Tan IIA) exerts an anticancer effect. Therefore, this study attempted to investigate whether GPER is involved in the inhibitory effect of Tan IIA on TNBC. We applied various databases and GO pathway analysis to predict the possible mechanism of Tan IIA. We identified 39 overlapping targets, including c-Jun, c-Fos, and caspase-3, and enriched cell cycle-related pathways. Next, we demonstrated the strong binding ability of Tan IIA to GPER by molecular docking assay. In the subsequent validation tests, Cell Counting Kit-8 (CCK8) assay showed that Tan IIA inhibited proliferation of MDA-MB-231 cells time and dose dependently without affecting normal cells. Using Transwell plate, flow cytometry, and Western blot assays, we showed that Tan IIA inhibited migration and induced apoptosis of MDA-MB-231 dose dependently. Importantly, protein expressions of GPER, epidermal growth factor receptor (EGFR), extracellular regulated protein kinases (ERK), c-Fos, and c-Jun were all decreased by Tan IIA dose dependently. Administration of GPER inhibitor partly abolished these effects. Furthermore, nuclear translocation of c-Fos and c-Jun as well as cell cycle-related proteins was downregulated by Tan IIA dose dependently. In summary, Tan IIA could inhibit the proliferation and migration of MDA-MB-231 cells and induce apoptosis, and the possible mechanism may be the regulation of GPER-mediated pathways, suggesting that GPER could be a therapeutic target for TNBC.

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.

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.

G protein-coupled estrogen receptor (GPER) was reported to be a potential target in the breast cancer therapy. This study aimed to illuminate the function of GPER and its mediated PI3K/AKT pathway in cryptotanshinone (CPT) inducing cell apoptosis and antiproliferation effect on GPER positive breast cancer MCF-7 cells. Cell proliferation was tested by MTT assay. Apoptosis rates were tested by Annexin V-FITC/PI double staining and the cell cycle was researched by flow cytometry. Autodock vina was applied to make molecular docking between CPT or estradiol and GPER. siRNA technique and GPER specific agonist G-1 or antagonist G-15 were applied to verify the mediated function of GPER. Apoptosis and cell cycle related proteins, as well as the key proteins on PI3K/AKT signaling pathway were detected by western blot. The results indicated that CPT could exert antiproliferation effects by arresting cell cycle in G2/M phase and downregulating the expression of cyclin D, cyclin B and cyclin A. Besides, apoptosis induced by CPT was observed. CPT might be a novel GPER binding compounds. Significantly, suppression of PI3K/AKT signal transduction by CPT was further increased by G-1 and decreased by G-15. The study revealed that the effect of antiproliferation and apoptosis treating with CPT on MCF-7 cells might be through the downregulation of PI3K/AKT pathway mediated by activated GPER.

目的 探讨2型糖尿病(T2DM)脐诊与中医证候、四诊信息的内在联系以及脐诊对于2型糖尿病的诊疗价值.方法 对90例T2DM患者进行中医证候判定、脐诊诊察及四诊信息调查,同时以90例正常成年人作为对照,录入并分析数据.结果 T2DM患者的肚脐大小与气虚积分、阳虚积分相关;肚脐的形状与大便的异常、带脉的功能相关.结论 肚脐的大小可反映气血的盈亏和脏腑的虚实变化,2型糖尿病患者多存在气血不足的情况,治疗应注重扶正;脐形可以判断带脉虚实,从带脉角度指导2型糖尿病的诊疗.

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.

Ethnopharmacological relevance: Si-Wu-Tang (SWT), a prestigious herbal formula from China, has been extensively used for centuries for female-related diseases. It has been documented that SWT has a significant inhibitory effect on non-triple-negative breast cancer (non-TNBC) cells. However, there has been limited comprehensive analysis of the targeted effects of the anticancer components of SWT and its exact biological mechanism. Aim of the study: This study aims to uncover the mechanism by which SWT treats non-TNBC by applying a network pharmacological method combined with experimental validation. Materials and methods: First, SWT compounds were collected from the Traditional Chinese Medicines Systems Pharmacology database (TCMSP) and The Encyclopedia of Traditional Chinese Medicine (ETCM), and then the targets related to SWT were obtained from the TCMSP and SwissTarget databases. Second, a target data set of non-TNBC proteins was established by using the Online Mendelian Inheritance in Man (OMIM), GeneCards and Gene Expression Omnibus (GEO) databases. Third, based on the overlap of targets between SWT and non-TNBC, a protein-protein interaction (PPI) network was built to analyse the interactions among these targets, which focused on screening for hub targets by topology. On these hub genes, we conducted a meta-analysis and survival analysis to screen the best match targets, ESR1, PPARG, CAT, and PTGS2, which had a strong correlation with the ingredients of SWT in our verification by molecular docking. In vitro experiments further proved the reliability of the network pharmacology findings. Finally, FunRich software and the ClusterProfiler package were utilized for the enrichment analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) data. Results: A total of 141 active ingredients and 116 targets of SWT were selected. GO enrichment analysis showed that the biological processes through which SWT acted against non-TNBC (FDR<0.01) mainly involved modulating energy metabolism and apoptosis. According to RT-qPCR and Western blotting, the mRNA and protein expression of ESR1, PPARG and PTGS2 were upregulated (P < 0.01), and the mRNA and protein levels of CAT were downregulated (P < 0.01), suggesting a multi-gene regulatory molecular mechanism of SWT against nontriple-negative breast cancer. Conclusions: This research explored the multi-gene pharmacological mechanism of action of SWT against nonTNBC through network pharmacology and in vitro experiments. The findings provide new ideas for research on the mechanism of action of Chinese medicine against breast cancer.

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:总结2型糖尿病肝郁气滞证的常见腹证特点,探讨不同腹证与四诊信息、中医证候的内在联系,挖掘辨证意义较大的典型腹证,提高临床辨证的准确性.方法:对北京中医药大学东直门医院门诊及住院治疗的90例2型糖尿病肝郁气滞证患者进行腹诊诊察及四诊信息调查,判定中医证候及积分,录入并分析数据.结果:常见腹证出现频率从高到低依次为心下痞硬、胸胁苦满、腹皮拘急、腹痛、心下痞;胸胁苦满、心下痞与肝郁气滞证的严重程度相关;胸胁苦满与舌边有浊沫存在相关性;心下痞硬、腹痛、胸胁苦满、心下冷凉与失眠多梦相关.结论:腹诊对于2型糖尿病的临床辨证论治有较大价值;胸胁苦满和心下痞可作为辅助2型糖尿病肝郁气滞证诊断的典型腹证;部分腹证与临床症状存在关联性,也为治疗原则的确立与疗效评估提供了思路方法.

桡骨茎突狭窄性腱鞘炎是由于反复的机械性摩擦而引起的慢性无菌性炎性改变,是临床常见病、多发病,临床主要是以腕部桡侧疼痛为主要症状,同时手指活动时可伴有弹响声,多与外伤、慢性劳损、寒冷刺激、遗传等因素相关,该病复发率高,且发病人群有低龄化趋势,目前治疗手段主要有推拿、针刺、艾灸、中药内服外用、手术、支具固定、局部封闭、微创等,临床应根据具体情况选用合适的治疗方法.