Comprehensive quality evaluation of Lysimachia christinae Hance via fingerprint, spectrum-effect relationship, and quantitative analyses of multiple components by single marker.

BACKGROUND: Lysimachia christinae Hance (LCH) is a traditional medicine used to treat gallstone disease and cholecystitis. Despite its known anti-inflammatory and choleretic effects, its quality has not been extensively evaluated. OBJECTIVE: In this study, we aimed to establish a reliable quality evaluation method for LCH via fingerprint, spectrum-effect relationship, and quantitative analyses of multicomponents by a single marker (QAMS). METHODS: First, the fingerprints and anti-inflammatory and choleretic activities of 14 LCH batches were determined. Then, the gray relation analysis method was used to analyze the peak areas of the fingerprint profile and pharmacodynamic data. Subsequently, the characteristic peaks were tentatively identified using high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Finally, rutin was selected as the internal reference material, and QAMS was used to analyze the LCH components. RESULTS: Pharmacodynamic experiments confirmed that LCH exerted anti-inflammatory and choleretic effects. Moreover, 15 flavonoids related to the anti-inflammatory and choleretic effects of LCH were identified. Notably, relative error percentage between the QAMS and external standard method was less than 5%. CONCLUSION: This study successfully established a comprehensive evaluation method for the qualitative and quantitative analyses of LCH.

Quantification of promoting efficiency and reducing toxicity of Traditional Chinese Medicine: A case study of the combination of Tripterygium wilfordii hook. f. and Lysimachia christinae hance in the treatment of lung cancer.

Traditional Chinese medicine (TCM) usually acts in the form of compound prescriptions in the treatment of complex diseases. The herbs contained in each prescription have the dual nature of efficiency and toxicity due to their complex chemical component, and the principle of prescription is usually to increase efficiency and reduce toxicity. At present, the studies on prescriptions have mainly focused on the consideration of the material basis and possible mechanism of the action mode, but the quantitative research on the compatibility rule of increasing efficiency and reducing toxicity is still the tip of the iceberg. With the extensive application of computational pharmacology technology in the research of TCM prescriptions, it is possible to quantify the mechanism of synergism and toxicity reduction of the TCM formula. Currently, there are some classic drug pairs commonly used to treat complex diseases, such as Tripterygium wilfordii Hook. f. with Lysimachia christinae Hance for lung cancer, Aconitum carmichaelii Debeaux with Glycyrrhiza uralensis Fisch. in the treatment of coronary heart disease, but there is a lack of systematic quantitative analysis model and strategy to quantitatively study the compatibility rule and potential mechanism of synergism and toxicity reduction. To address this issue, we designed an integrated model which integrates matrix decomposition and shortest path propagation, taking into account both the crosstalk of the effective network and the propagation characteristics. With the integrated model strategy, we can quantitatively detect the possible mechanisms of synergism and attenuation of Tripterygium wilfordii Hook. f. and Lysimachia christinae Hance in the treatment of lung cancer. The results showed the compatibility of Tripterygium wilfordii Hook. f. and Lysimachia christinae Hance could increase the efficacy and decrease the toxicity of lung cancer treatment through MAPK pathway and PD-1 checkpoint pathway in lung cancer.

Aqueous Extract of Lysimachia christinae Hance Prevents Cholesterol Gallstone in Mice by Affecting the Intestinal Microflora.

With changes in human dietary patterns, the proportion of high-fat and high-cholesterol foods in the daily diet has increased. As a result, the incidence rate of cholelithiasis is increasing rapidly. Many studies have reported on the crucial role that the intestinal microflora plays in the progression of gallstones. Although the whole herb of Lysimachia christinae, a traditional Chinese medicine, has long been extensively used as a remedy for cholelithiasis in China, its effects on the intestinal microflora remain unknown. Hence, in this study, we investigated the ability of the aqueous extract of L. christinae (LAE) to prevent cholesterol gallstones (CGSs) in model animals by affecting the intestinal microflora. The effects of LAE on body weight, serum lipid profile, visceral organ indexes, and histomorphology were studied in male C57BL/6J mice, which were induced by a lithogenic diet. After the 8-week study, CGSs formation was greatly reduced after LAE treatment. LAE also reduced body weight gain and hyperlipidemia and restored the histomorphological changes. Moreover, the intestinal microflora exhibited significant variation. In the model group fed the lithogenic diet, the abundances of the genera unclassified Porphyromonadaceae, Lactobacillus and Alloprevotella decreased, but in contrast, Akkermansia dramatically increased compared with the control check group, which was fed a normal diet; the administration of LAE reversed these changes. These results imply that L. christinae can be considered an efficient therapy for eliminating CGSs induced by a high-fat and high-cholesterol diet, which may be achieved by influencing the intestinal microflora.

Lysimachia christinae Hance as an anticancer agent against breast cancer cells.

Breast cancer is the most common cancer in women, and metastasis is the leading cause of death in breast cancer patients. Although chemoprevention is widely employed to treat breast cancer, anticancer drugs can cause significant adverse effects. Lysimachia christinae Hance (LH) is a traditional Chinese medicinal plant with diverse therapeutic effects. However, its potential anticancer activity has not been fully investigated in breast cancers to date. Using high-performance liquid chromatography-mass spectrometry, we found that the main constituent of LH extract (LHE) was rutin. Our results indicated that LHE or rutin markedly decreased the proliferation and viability of estrogen receptor (ER)-positive MCF-7 and ER-negative HCC38 human breast cancer cells. LHE treatment induced morphological changes in apoptotic nuclei using 4',6-diamidino-2-phenylindole (DAPI) staining. Annexin V-fluorescein isothiocyanate (FITC) propidium iodide (PI) staining assay revealed that apoptosis significantly increased in both breast cancer cell types after LHE treatment. Additionally, the expression of poly (ADP-ribose) polymerase (PARP), Bcl-2, and phospho-Akt decreased, while that of cleaved PARP and p53 increased, in both cell types. Furthermore, LHE treatment inhibited epithelial-mesenchymal transition (EMT). LHE treatment significantly upregulated E-cadherin level in MCF-7 and HCC38 cells, while vimentin level was downregulated in HCC38 cells. In addition, transwell and wound-healing assays revealed that LHE or rutin inhibited breast cancer cell migration. Overall, these findings demonstrate that LHE is a promising therapeutic agent that acts by promoting apoptosis and reducing cell proliferation, EMT, and cell migration in ER-positive and ER-negative breast cancer cells.

The Effects of Lysimachia christinae Hance Extract Fractions on Endothelium-Dependent Vasodilatation.

BACKGROUND: Endothelium-dependent dilatation is a predictor for vascular function. NADPH oxidase-derived O2- can inactivate nitric oxide and induce vascular injury. METHOD: The crude ethanolic extract of Lysimachia christinae Hance were separated out 4 fractions of different olarities by petroleum ether, ethyl acetate, n-butanol (NB), and aqueous. The endothelial integrity was appraised by vascular tension measurement. Dihydroethidium was utilized to observe the vascular reactive oxygen species (ROS) production. Western-blot was adopted to detect protein expression. RESULTS: Among the 4 fractions of L. christinae Hance, the NB fraction showed the most potent capacity of promoting endothelium-dependent vascular relaxation and inhibiting ROS formation in aortic rings, which were likely attributed by suppressing the expression of NAD(P)H oxidase subunit (gp91phox, p47phox, and p67phox) and enhancing the phosphorylation of endothelial NOS in vascular tone. CONCLUSIONS: These results suggest that the NB fraction possess the strongest vascular pharmacological activities among the crude ethanolic extract of L. christinae Hance, which may help us for purifying bioactive constituents and discovering new drugs from this herb in future.

Detoxication mechanisms of Radix Tripterygium wilfordii via compatibility with Herba Lysimachia christinae in S180-bearing mice by involving Nrf2.

The combined administration between Radix Tripterygium wilfordii Hook F (LGT) and Herba Lysimachia christinae Hance (JQC) belongs to mutual detoxication compatibility of seven emotions in traditional Chinese medicine (TCM) theory. However, until now, the compatibility detoxication mechanisms remain unknown. The present study was undertaken to observe detoxication mechanisms of LGT through compatibility with JQC in tumor-bearing mice by involving NF-E2-related factor 2 (Nrf2)-mediated antioxidant defenses. In addition, influence of compatibility on antitumor activity was also investigated here. Our results demonstrated that compatibility with JQC administration significantly reversed LGT-elevated serum alanine/aspartate transaminase (ALT/AST) levels and alleviated hepatocytes' swelling or degeneration damage, and at the ratio 2/1 (LGT/JQC) produced the strongest detoxication effect. Besides, compatibility with JQC administration reversed not only LGT-elevated hepatic malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α) but also the LGT lowered GSH, glutathione-s transferase (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and interleukin (IL)-10 levels. Furthermore, compatibility with JQC administration significantly up-regulated protein expression of Nrf2 and mRNA expression of it regulated downstream antioxidant genes such as heme oxygenase-1 (HO-1), NAD(P)H: quinone oxidoreductase-1 (NQO1), and glutamate cysteine ligase catalytic subunit (GCLC). In addition, compatibility with JQC further decreased LGT-decreased tumor weight and at the ratio 2/1 (LGT/JQC) also exerted the strongest synergistic effect. Collectively, through compatibility with JQC exerted detoxication effect on LGT-induced hepatotoxicity and the mechanisms could be at least partly attributed to up-regulation of Nrf2 and its downstream signals, thereby enhancing antioxidant defenses, and inhibiting lipid peroxidation, oxidative stress, and inflammation. Additionally, at the ratio 2/1 (LGT/JQC) exerted the strongest effects on both detoxication and synergism.

Evaluation of the antioxidant and endothelial protective effects of Lysimachia christinae Hance (Jin Qian Cao) extract fractions.

BACKGROUND: Lysimachia christinae Hance is a traditional Chinese medicine with diuretic, detumescent, and detoxifying effects. Our aimed to optimize the extraction protocol to maximize the yield of flavonoids from Lysimachia christinae Hance, and evaluate the pharmacological activities of four fractions, namely, petroleum ether (PE), ethyl acetate (EA), n-butanol (NB), and aqueous (AQ) fractions, of the ethanolic extract of Lysimachia christinae Hance. METHODS: The flavonoid monomers in the crude extract were characterized via high performance liquid chromatography (HPLC), were used as markers for extract quality control and standardization. The total flavonoid, total phenolic, and total polysaccharide contents of each fraction were determined by spectrophotometry. Further, the in vitro free radical (diphenylpicrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), superoxide, and hydroxyl radicals) scavenging activities, and antioxidant capacity in endothelial cells were evaluated for each fraction. RESULTS: After optimizing the extraction protocol to maximize the total flavonoid yield from L. christinae Hance, the NB fractions had the highest total flavonoid (39.4 ± 4.55 mg RE/g), total phenolic (41.1 ± 3.07 mg GAE/g) and total polysaccharide (168.1 ± 7.07 mg GE/g); In addition, the NB fraction of the ethanolic extract of L. christinae Hance reveal the strongest radical-scavenging activity, antioxidant activity and protective effects against H2O2-induced injury in HUVECs. CONCLUSIONS: Among the four fractions of L. christinae Hance, the NB fraction showed the most potent antioxidant and endothelial protective effects, which may be attributed to its high flavonoid, phenolic contents and optimal portfolio of different active ingredients of NB fractions of the ethanolic extract of L. christinae Hance. This study might improve our understanding of the pharmacological activities of L. christinae Hance, thereby facilitating its use in disease prevention and treatment.

Anti-tumor components from Lysimachia christinae Hance / 军事医学

Objective To isolate and identify anti-tumor constituents from Lysimachia christinae Hance.Methods The compounds were isolated and purified by chromategraphy on AB-8 macroporous resin, silica gel, Sephadex LH-20, ODS and preparative HPLC.Their structures were elucidated using chemical and spectroscopic methods.All thecompounds obtained were evaluated on their growth inhibitory effects in vitro using the MTT method.Results Eleven compounds were isolated from the whole plant of L.christinae Hance and were identified as 3,3'-dimethoxy-6,6'-di[(Z)-pentadec-10-en-1-yl]-[1,1'-bi(cyclohexane)]-3,3',6,6'-tetraene-2,2',5,5'-tetraone(1),physcion(2),munjistin(3), 9,16-dioxooctadec-10,12,14-trienoic acid(4),cyclamiretinA-3-O-{β-D-xylopyranosyl-(1→2)-β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-α-L-arabinopyranoside(5),lysikoianoside(6),quercetin-3-O-glucopyranoside(7),isorhamnetin3-O-β-D-galactopyranoside(8),3',4',5',5,7-pentahydroxyflavone(9), rutin(10), and quercetin(11).The screening results of antitumor activity in vitro showed that IC50 of compound 5 for human cervical carcinoma cells HeLa, human osteosarcoma cells U20S, human lung adenocarcinoma cells PC-9,and human colorectal carcinoma cells CT-26 was 2.29, 1.22, 1.43, and 1.29 μmol/L respectively.Conclusion Compound 1 is a new compound and compounds 2-4 are isolated from the plants of Lysimachia for the first time.Compound 5 shows obvious inhibitory effects on tumor cell growth in vitro.

Comparison on HPLC fingerprints of Lysimachia christinae and Desmodium styracifolium / 中草药

Objective: The differences in the compounds between Lysimachia christinae and Desmodium styracifolium were compared in order to provide the references for pharmacodynamic differences of them. Methods: The RP-HPLC-UV and RP-HPLC-ELSD fingerprints of L. christinae and D. styracifolium were established, and the common peaks of two kinds of fingerprints were compared. Results: The HPLC-UV fingerprints of L. christinae were obviously different from the HPLC-ELSD fingerprints, and the strong peaks detected in UV were not the same as what detected in ELSD, so were those of D. styracifolium. There were 14 and 5 common peaks, respectively in the HPLC-UV and HPLC-ELSD fingerprints of L. christinae, while 28 and 18 common peaks, respectively in the HPLC-UV and HPLC-ELSD fingerprints of D. styracifolium. The similarities of chromatograms of D. styracifolium were better than those of L. christinae. Four common peaks were identified in the HPLC-UV fingerprints of L. christinae and D. styracifolium, and two common peaks in their HPLC-ELSD fingerprints. Conclusion: It is better using HPLC-ESLD to establish the fingerprints of L. christinae and D. styracifolium. The chemical constituents of L. christinae were significantly different between the different batches. However, little variation was found in the chromatograms between the different batches of D. styracifolium. The two plants may contain two same compounds with high content, which may have the function as the substantial bases of their treatments for the same indication. However, they may have their special efficacies.

Genetic diversity of Lysimachia christinae by ISSR / 中国药学杂志

OBJECTIVE: To analyze the genetic diversity of Lysimachia christinae Hance germplasm resources from 36 different places of origin with ISSR DNA markers. METHODS: Ten primers screened from 45 ISSR primers wrea mplified with PCR and the amplification products were examined by 1.5% agarose gel electrophoresis. POPGENE32 software was used for statistical analysis of genetic parameters and UPGMA method (NTSYS2.10 software) for cluster analysis and dendrogram constructing. RESULTS: A total of 91 sites were amplified from the 10 ISSR primers, of which 80 sites were polymorphic loci. The percentage of total polymorphic loci (PBB) was 87.91%. The number of observed alleles (Na) was 1.879 1 and the number of effective alleles was number (Ne) 1.381 7. The Nei's genetic diversity index (He) was 0.239 0 and the Shannon information index (I) was 0.374 5. The genetic similarity coefficient of the 36 germplasm resources varied from 0.69 to 0.97 and the 36 materials couldbe divided into six categories at the similarity coefficient of 0.76. Clustering results showed that the samples within small geographic range clustered together in the dendrogram, indicating the nearer genetic relationships of them. Meanwhile, the samples from different provinces or cities could not be differentiated from each other through cluster analysis. CONCLUSION: The germplasm resources of Lysimachia christinae have rich genetic diversities. The genetic diversities of Lysimachia christinae have no significant correlation with their geographic distribution.

Hepatoprotective effects of Lysimachia christinae Hanceextracts on acute liver injury induced by tripterygium glycosides in mice / 中国药学杂志

OBJECTIVE: To investigate the hepatoprotective effects of Lysimachia christinae Hance against acute liver injury induced by tripterygium glycosides (TG) in vivo and the related mechanism for the first time. METHODS: The mice were administered ethanol extract of L. christinae (JE), water extract of L. christinae (JW), and bifendate by ig for seven consecutive days. 12 h after the last dose, the mice were orally given TG 270 mg · kg-1 except those in the normal group. Serum and liver tissue samples were collected at 18 h after TG treatment. Besides, the amounts of quercetin and kaempferol in JE and JW were analyzed by high-performance liquid chromatography (HPLC). RESULTS: TG-induced elevated serum alanine transferase(ALT) and aspartate transaminase (AST) activities were significantly reduced by JE(100, 200 mg · kg-1) in dose-dependent manners, but not by JW. Further analysis demonstrated that lipid peroxidation(LPO) level significantly decreased, while superoxide dismutase(SOD) and catalase(CAT) activities increased in livers of JE-treated mice. Besides, the amounts of quercetin and kaempferol were 9.8 and 8.9 mg · g-1 in JE, and 2.8 and 1.9 mg · g-1 in JW, respectively. CONCLUSION: This study for the first time demonstrates that L. christinae can protect against TG-induced acute liver injury in dose-dependent manners in vivo, the potential mechanism may be related to inhibiting liver oxidative stress injury, and the hepatoprotective activity may be correlated with the contents of quercetin and kaempferol. Copyright 2013 by the Chinese Pharmaceutical Association.