Effect of Sparganii Rhizoma-Curcumae Rhizoma-medicated serum on proliferation, apoptosis, and migration of human ectopic endometrial stromal cells: based on JAK2/STAT3 signaling pathway / 中国中药杂志

Based on the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signaling pathway, this study investigated the effect of medicated serum of Sparganii Rhizoma(SR) and Curcumae Rhizoma(CR) on the proliferation, apoptosis, migration, and secretion of inflammatory factors of ectopic endometrial stromal cells(ESCs). Specifically, human ESCs were primary-cultured. The effect of different concentration(5%, 10%, 20%) of SR-, CR-, and SR-CR combination-medicated serum, and AG490 solution(50 μmol·L~(-1)) on the proliferation of ESCs was detected by methyl thiazolyl tetrazolium(MTT) assay, and the optimal dose was selected accordingly for further experiment. The cells were classified into normal serum(NS) group, SR group(10%), CR group(10%), combination(CM) group(10%), and AG490 group. The apoptosis level of ESCs was detected by flow cytometry, and the migration ability was examined by wound healing assay. The secretion of interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α was determined by enzyme-linked immunosorbent assay(ELISA). The protein levels of cysteinyl aspartate specific protei-nase-3(caspase-3), B-cell lymphoma(Bcl-2), and Bcl-2-associated X protein(Bax) and the levels of phosphorylated(p)-JAK2 and p-STAT3 were detected by Western blot. The results showed that the viability of ESCs cells was lowered in the administration groups compared with the blank serum group(P<0.01), especially the 10% drug-medicated serum, which was selected for further experiment. The 10% SR-medicated serum, 10% CR-medicated serum, and 10% CM-medicated serum could increase the apoptosis rate(P<0.01), up-regulate the protein expression of caspase-3 and Bax in cells(P<0.05 or P<0.01), down-regulate the expression of Bcl-2(P<0.01), decrease the cell migration rate(P<0.05 or P<0.01), and reduce the secretion levels of IL-1β, IL-6, and TNF-α(P<0.05 or P<0.01), and levels of p-JAK2 and p-STAT3(P<0.05 or P<0.01). Compared with the SR and CR groups, CM group showed low cell viability(P<0.01), high protein expression of caspase-3 and Bax(P<0.05 or P<0.01), and low protein expression of Bcl-2 and p-JAK2(P<0.05). After incubation with CM, the apoptosis rate was higher(P<0.05) and the migration rate was lower(P<0.01) than that of the CR group. The p-STAT3 protein level of CM group was lower than that of the RS group(P<0.05). The mechanism of SR, CR, and the combination underlying the improvement of endometriosis may be that they blocked JAK2/STAT3 signaling pathway, inhibited ESC proliferation, promoted apoptosis, weakened cell migration, and reduced the secretion of inflammatory factors. The effect of the combination was better than that of RS alone and CR alone.

Discovery and identification of quality markers of Sparganii Rhizoma based on zebrafish thrombosis model / 中草药·英文版

Objective: The aim of the present study was to determine the quality marker (Q-Markers) of Sparganii Rhizoma against thrombus through an integration of investigations on its antithrombotic effect, content determination and spectrum-effect correlation analysis. Methods: Based on the concept of Q-Marker, Sparganii Rhizoma was investigated for the identification of chemical component. The pharmacological effects on arachidonic acid-induced thrombosis in zebrafish were also investigated. The material basis in ethanol extract was determined by HPLC-UV. Furthermore, the potential Q-Markers were analyzed and predicted according to the effect-chemical correlation analysis. Finally, the anti-thrombotic Q-Markers were verified through the anti-thrombotic test of monomer components. Results: The model of thrombosis zebrafish was established with larvae exposed to 100 µmol/L arachidonic acid for 1 h. Nine ingredients in Sparganii Rhizoma were identified as 5-hydroxymethylfurfural, vanillic acid, ferulic acid, p-hydroxybenzaldehyde, p-hydroxybenzoic acid, vanillin, protocatechuic acid, p-coumaric acid and isoferulic acid. According to the determination effect of zebrafish thrombosis model and HPLC content analysis results, all the other contents present positive correlation except 5-hydroxymethylfurfural, and the P values of three representative potential Q-Markers (ferulic acid, protocatechuic acid and p-coumaric acid) were 0.002, 0.001 and 0.026, respectively. Conclusion: Sparganii Rhizoma showed a dose-dependent effect on the recovery of reducing cardiac red blood cell on zebrafish model. Three phenolic acids (ferulic acid, protocatechuic acid and p-coumaric acid) were proved to possess the anti-thrombotic effects which could be regarded as the potential Q-Markers for quality assessment of Sparganii Rhizoma.

Phenols and Organic Acids Constituents in Water Decoction of Curcumae Rhizoma-Sparganii Rhizoma Herbal Pair and Single Herb / 中国实验方剂学杂志

Objective:To investigate constituents containing phenolic hydroxyl or carboxylic acid （excluding diarylheptanoids） from Curcumae Rhizoma and Sparganii Rhizoma herbal pair and single herb. Method:Multiple chromatographic separation techniques， including silica gel，MCI gel and Sephadex LH-20 gel， were employed to isolate and purify the compounds. Their structures were identified by means of the nuclear magnetic resonance （NMR），mass spectrometry （MS） and physicochemical properties. Constituents were quickly analyzed by UPLC-LTQ-Orbitrap-MSn，and scanned in positive and negative ion modes. Result:These compounds were determined as trans-p-hydroxycinnamic acid（1），vanillic acid（2），protocatechuic acid（3），fumalic acid（4），succinic acid（5），succinic acid monomethyl ester（6），docosanoic acid（7），azelaic acid（8） and p-hydroxybenzaldehyde（9）. Forty compounds were speculated by comparing mass spectrometry data，retention times of some compounds and reference materials，including 14 phenols and 26 organic acids. Among the compounds of herbal pair，8 phenols and 22 organic acids in Sparganii Rhizoma as well as 11 phenols and 13 organic acids in Curcumae Rhizoma were identified. Cleavage pathways of main compounds were described. Conclusion:There are abundant phenols and organic acids in Curcumae Rhizoma and Sparganii Rhizoma herbal pair and single herb. The results enrich pharmacodynamic material basis of Curcumae Rhizoma and Sparganii Rhizoma herbal pair.

Molecular Mechanism of Anti-thrombosis and Anti-platelet Aggregation of Sparganii Rhizoma Based on Integrated Pharmacological Platform / 中国实验方剂学杂志

Objective: To study on the molecular mechanism of anti-platelet aggregation and anti-thrombosis of Sparganii Rhizoma. Method: Based on the data information and analysis functions of traditional Chinese medicine(TCM) database,TCM composition database and target database in TCM platform for integration of pharmacology,information of chemical compositions in Sparganii Rhizoma was retrieved,interaction network between potential targets of Sparganii Rhizoma and disease targets was built,and the key targets were enriched and calculated,the gene functions and pathways were analyzed,multidimensional relationship network of " herbal medicines-chemical components-key targets-key pathways" was built. Result: Active ingredients of Sparganii Rhizoma mainly included flavonoids and phenolic acids,such as mountain kaempferol,sanleng acid,linoleic acid,etc.Anti-thrombosis involved 202 key targets,including protein kinase Cδ(PRKCD),glucose kinase(GCK),(PRKAA2),adenosine ribonucleotide activated protein kinase α-2 catalytic subunit,etc;and it was related to the endocrine system,circulatory system,neurodegenerative diseases and other related biological processes and signal pathways.Anti-platelet aggregation involved 136 key targets,including PRKCD,cytochrome C oxidase protein 7C(COX7C),GCK,etc;and it was involved in Parkinson's disease,circulatory system,neurodegenerative diseases and other related biological processes and signal pathways. Conclusion: Sparganii Rhizoma regulates vascular endothelial cell adhesion molecule expression and platelet mitochondrial function mediated apoptosis of platelets mainly through regulating neurotransmitter activity in the central nervous system,in order to exert antithrombotic effect and anti-platelet aggregation effect.

Transcriptional profiling of uterine leiomyoma rats treated by a traditional herb pair, Curcumae rhizoma and Sparganii rhizoma

The aim of this study was to elucidate the concise effects of a traditional herb pair, Curcumae rhizoma-Sparganii rhizoma (CRSR), on uterine leiomyoma (UL) by analyzing transcriptional profiling. The UL rat model was made by intramuscular injection of progesterone and gavage administration of diethylstilbestrol. From 11 weeks of the establishment of the model, rats of the UL+CRSR group were gavaged daily with CRSR (6.67 g/kg). The serum concentrations of progesterone (P) and estradiol (E2) were determined by radioimmunoassay, the uterine index was measured by caliper measurement, and the pathological status was observed by hematoxylin and eosin stain. Gene expression profiling was checked by NimbleGen Rat Gene Expression Microarrays. The results indicated that the uterine mass of UL+CRSR rats was significantly shrunk and serum P and E2 levels significantly reduced compared to UL animals and nearly to the level of normal rats. Results of microarrays displayed the extensive inhibition of CRSR upon the expression of proliferation and deposition of extracellular matrix (ECM)-related genes, and significantly regulated a wide range of metabolism disorders. Furthermore, CRSR extensively regulated key pathways of the UL process, such as MAPK, PPAR, Notch, and TGF-β/Smad. Regulation of the crucial pathways for the UL process and ECM metabolism may be the underlying mechanisms of CRSR treatment. Further studies will provide clear clues for effectively treating UL with CRSR.

Comparative chemical characters of Sparganii Rhizoma from different regions in China / 中草药·英文版

Objective: To compare the chemical characters of Sparganii Rhizoma from different areas via chromatographic analysis and to establish a sensitive LC/MS method for quality assessment of Sparganii Rhizoma. Methods: Under the optimised HPLC-PDA chromatographic conditions, twenty batches of Sparganii Rhizoma were analyzed by HPLC fingerprints. Principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (OPLS-DA) and hierarchical cluster analysis (HCA) were performed based on all peak areas of Sparganii Rhizoma fingerprints. Meanwhile, part of common peaks were subsequently quantified by UFLC-QTRAP-MS. Results: The similarity values of HPLC fingerprints fluctuated in a wide range of 0.511–0.973, which showed variable differences of chemical characters among Sparganii Rhizoma from twenty habitats. PCA, OPLS-DA and HCA indicated that samples could be divided into five groups with different chemical characters, which generally corresponded with their geographical distributions. A total of 31 peaks in HPLC fingerprints were marked, and eight of them were identified and quantified. The quantitative result was generally in agreement with the classifications based on HPLC fingerprints, which indicated that Sparganii Rhizoma samples from eastern China mostly contained more contents including phenolic acids and flavonoids. Conclusion: This study not only proved that there were relationships between geographic distributions and internal chemical compositions of plants, which could provide evidence to the traditional Chinese medicine concept “geo-authentic” but also supplied a sensitive and rapid simultaneous quantitive method for the quality estimation of Sparganii Rhizoma.

Effects of active components in compatible solution of Sparganii Rhizoma-Curcumae Rhizoma on pelvic adhesion in rats with chronic pelvic inflammatory disease / 中成药

AIM To study the influence and action mechanism of effective components in compatible solution of Sparganii Rhizoma-Curcumae Rhizoma on pelvic adhesions of model rats with chronic pelvic inflammatory disease.METHODS Laboratory rats were randomly assigned into six groups,a sham-operated group,a model group,a positive Fukeqianjin tablets group,compatible solution groups (40 g/kg high-dose group,20 g/kg medi um-dose group,and 10 g/kg low-dose group).Except rats of the sham-operated group,those in the other groups were induced into models of chronic pelvic inflammatory disease by mechanical bacterial strain implantation,after which a 20 d corresponding drug administration by oral gavage for each group was initiated.After completion of the last drug administration,Verco criterion was taken as the reference to evaluate pelvic adhesions,HE staining was performed on uterine tissues for pathological scoring,ELASA method was applied to measuring serum IL-1β and TNF-α levels,and Western blot method was used to detect the expression levels of proteins FGF-2 and IGF-1 in uterine tissues.RESULTS Compared with the model group,the high-dose compatible solution group had a much sharper fall in Verco scores (P ＜ 0.05),and remarkably decreased serum IL-1β and TNF-α levels (P ＜ 0.01);both high-dose and medium-dose groups manifested with notably alleviated degenerative necrosis and lowered scores of chronic inflammation infiltration and epithelial hyperproliferation (P ＜ 0.01 or P ＜ 0.05),and a distinct reduction in the expression of proteins FGF-2 and IGF-1 (P ＜ 0.05,P ＜ 0.01).CONCLUSION The significant improvement of the chronic pelvic inflammatory disease in model rats due to the effective components in the compatibile solution of Sparganii Rhizoma-Curcumae Rhizoma may be associated with inhibited release of inflammatory mediators IL-1β and TNF-α,and the reduced expression of proteins FGF-2 and IGF-1 as well.

Research progress in inhibition of Chinese materia medica with breaking blood stasis and resolving mass on tumor metastasis / 中草药

Activating blood and dissolving stasis is one of the important principles of clinical anti-tumor by traditional Chinese medicine and the anti-tumor effect of the blood-activating and stasis-resolving drugs has been confirmed by a large number of studies. But in recent years, some experimental studies have found that some blood-activating and stasis-resolving drugs could promote tumor metastasis. And then the clinical application of this kind of drugs has suffered from many controversial problems. Chinese materia medica (CMM) with breaking blood stasis and resolving mass (Poxue herbs) is a kind of blood-activating and stasis-resolving drugs which display the severe effect. Nowadays some researchers confirmed its obvious antineoplastic effect and its prominent anti-tumor metastasis effect, which indicated in recent studies. In this article, we summarized the anti-tumor metastasis effect and mechanism of the Poxue herbs like Curcumae Rhizoma, Sparganii Rhizoma, large blister Beetle, leech, and its effective components in detail. And we also put forward that Poxue herbs are the ones which have the anti-tumor metastasis effect among blood-activating and stasis-resolving drugs, and it valued with significant clinical application and development. It is also believed that the Poxue herbs could inhibit the tumor metastasis which is related with aerobic glycolysis and can provide the theoretical direction to guide clinical application.

Effect of different vinegar processing technologies on dissolution of effective composition groups in Sparganii Rhizoma / 中草药

Objective: To study the dissolving yield and ratio fluctuation of Sparganii Rhizoma effective composition groups, including total flavonoids, total alkaloid, and active Al3+, when processed with different vinegar processing technologies. Methods: L9(34) orthogonal test design was used with acetic acid content (A), vinegar ratio (B), soaking time (C), and frying temperature (D) as influencing factors. The differences of dissolving yield and ratio of 80% methanol extract, total flavonoids, total alkaloid, and active Al3+ of processed Sparganii Rhizoma under different vinegar processing technologies were compared. Results: Compared with crude Sparganii Rhizoma, the processed products with a little acetic acid had enhanced the dissolving yield of 80% methanol extract. Moreover, the processed Sparganii Rhizoma with vinegar had effectively enhanced the dissolving yield and ratio of total flavonoids, total alkaloid, and active Al3+. The dissolving yield and ratio of total flavonoids were greatly impacted by frying temperature. Meanwhile, the alkaloid dissolving yield and ratio were most influenced by acetic acid content. The products processed with abundant acetic acid had obviously reduced the dissolving yield of 80% methanol extract, but had increased the alkaloid and active Al3+ dissolving ratio in the extract. Conclusion: The processed Sparganii Rhizoma with vinegar could obviously affect the dissolving yield and ratio of different pharmacodynamic compositions in Sparganii Rhizoma. Suitable processed products should be adopted when Sparganii Rhizoma is used in the different treatments.