Chemical and bioactive comparison of Panax notoginseng root and rhizome in raw and steamed forms.

BACKGROUND: The root and rhizome are historically and officially utilized medicinal parts of Panax notoginseng (PN) (Burk.) F. H. Chen, which in raw and steamed forms are used differently in practice. METHODS: To investigate the differences in chemical composition and bioactivities of PN root and rhizome between raw and steamed forms, high-performance liquid chromatography analyses and pharmacologic effects evaluated by tests of anticoagulation, antioxidation, hemostasis, antiinflammation, and hematopoiesis were combined. RESULTS: With the duration of steaming time, the contents of ginsenosides Rg1, Re, Rb1, Rd, and notoginsenoside R1 in PN were decreased, while those of ginsenosides Rh1, 20(S)-Rg3, 20(R)-Rg3, Rh4, and Rk3 were increased gradually. Raw PN samples steamed for 6 h at 120°C with stable levels of most constituents were used for the subsequent study of bioeffects. Raw PN showed better hemostasis, anticoagulation, and antiinflammation effects, while steamed PN exhibited stronger antioxidation and hematopoiesis activities. For different parts of PN, contents of saponins in PN rhizome were generally higher than those in the root, which could be related to the stronger bioactivities of rhizome compared with the same form of PN root. CONCLUSION: This study provides basic information about the chemical and bioactive comparison of PN root and rhizome in both raw and steamed forms, indicating that the change of saponins may have a key role in different properties of raw and steamed PN.

Quality Evaluation of Gastrodia Elata Tubers Based on HPLC Fingerprint Analyses and Quantitative Analysis of Multi-Components by Single Marker.

Gastrodia elata (G. elata) tuber is a valuable herbal medicine used to treat many diseases. The procedure of establishing a reasonable and feasible quality assessment method for G. elata tuber is important to ensure its clinical safety and efficacy. In this research, an effective and comprehensive evaluation method for assessing the quality of G. elata has been developed, based on the analysis of high performance liquid chromatography (HPLC) fingerprint, combined with the quantitative analysis of multi-components by single marker (QAMS) method. The contents of the seven components, including gastrodin, p-hydroxybenzyl alcohol, p-hydroxy benzaldehyde, parishin A, parishin B, parishin C, and parishin E were determined, simultaneously, using gastrodin as the reference standard. The results demonstrated that there was no significant difference between the QAMS method and the traditional external standard method (ESM) (p > 0.05, RSD < 4.79%), suggesting that QAMS was a reliable and convenient method for the content determination of multiple components, especially when there is a shortage of reference substances. In conclusion, this strategy could be beneficial for simplifying the processes in the quality control of G. elata tuber and giving references to promote the quality standards of herbal medicines.

Steamed Panax notoginseng Attenuates Anemia in Mice With Blood Deficiency Syndrome via Regulating Hematopoietic Factors and JAK-STAT Pathway.

Panax notoginseng (Burk.) F. H. Chen is a medicinal herb used to treat blood disorders since ancient times, of which the steamed form exhibits the anti-anemia effect and acts with a "blood-tonifying" function according to traditional use. The present study aimed to investigate the anti-anemia effect and underlying mechanism of steamed P. notoginseng (SPN) on mice with blood deficiency syndrome induced by chemotherapy. Blood deficiency syndrome was induced in mice by cyclophosphamide and acetylphenylhydrazine. A number of peripheral blood cells and organs (liver, kidney, and spleen) coefficients were measured. The mRNA expression of hematopoietic function-related cytokines in the bone marrow of mice was detected by RT-qPCR. The janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway was screened based on our previous analysis by network pharmacology. The expression of related proteins and cell cycle factors predicted in the pathway was determined by Western blot and RT-qPCR. SPN could significantly increase the numbers of peripheral blood cells and reverse the enlargement of spleen in a dose-dependent manner. The quantities of related hematopoietic factors in bone marrow were also increased significantly after SPN administration. SPN was involved in the cell cycle reaction and activation of immune cells through the JAK-STAT pathway, which could promote the hematopoiesis.

Identification of anti-inflammatory components of raw and steamed Panax notoginseng root by analyses of spectrum-effect relationship.

To identify bioactive components of Panax notoginseng (PN) roots in raw and steamed forms, chemometrics analyses including bivariate correlation analysis and partial least squares regression were used to establish the relationships between the chromatograms and anti-inflammatory effect of PN samples. The chemical fingerprints of PN were obtained by high performance liquid chromatography. The anti-inflammatory activity was evaluated by the TNF-α and IL-6 inhibiting test. According to the results, raw PN (RPN) displayed a stronger anti-inflammatory effect compared with steamed PN (SPN) samples. Chemometrics analyses showed that notoginsenoside R1, ginsenosides Rg1, Re and Rb1 were the major anti-inflammatory components of RPN, whereas ginsenosides 20(S)-Rg3 was the active one of SPN treating inflammation. The pharmacologic verification was consistent with the predicted results. In addition, the contents of anti-inflammatory components in RPN were higher than those in SPN, which might be the reason that RPN exhibited stronger effect on the inflammation induced by lipopolysaccharide in RAW264.7 cells. These findings could be beneficial for choosing differential markers for the quality control of RPN and SPN.

Optimisation of Ethanol-Reflux Extraction of Saponins from Steamed Panax notoginseng by Response Surface Methodology and Evaluation of Hematopoiesis Effect.

The present study aims to optimize the ethanol-reflux extraction conditions for extracting saponins from steamed Panax notoginseng (SPN). Four variables including the extraction time (0.5⁻2.5 h), ethanol concentration (50⁻90%), water to solid ratio (W/S, 8⁻16), and times of extraction (1⁻5) were investigated by using the Box-Behnken design response surface methodology (BBD-RSM). For each response, a second-order polynomial model with high R² values (>0.9690) was developed using multiple linear regression analysis and the optimum conditions to maximize the yield (31.96%), content (70.49 mg/g), and antioxidant activity (EC50 value of 0.0421 mg/mL) for saponins extracted from SPN were obtained with a extraction time of 1.51 h, ethanol concentration of 60%, extraction done 3 times, and a W/S of 10. The experimental values were in good consistency with the predicted ones. In addition, the extracted SPN saponins could significantly increase the levels of blood routine parameters compared with the model group (p < 0.01) and there was no significant difference in the hematopoiesis effect between the SPN group and the SPN saponins group, of which the dose was 15 times lower than the former one. It is suggested that the SPN saponins extracted by the optimized method had similar functions of "blood tonifying" at a much lower dose.

Promotion of quality standard of Chinese herbal medicine by the integrated and efficacy-oriented quality marker of Effect-constituent Index.

BACKGROUND: Multiple constituents have been applied currently as markers to control the quality of Chinese herbal medicine (CHM). However, those constituents are isolated from each other, failed to present their contribution differences to the bioeffect of CHM. Besides, a CHM for different clinic uses is often controlled by the same quality marker (Q-marker), which cannot correlate its efficacies differentially. PURPOSE: The study aims to promote the quality standard of CHM by the integrated and efficacy-oriented Q-marker of Effect-constituent Index (ECI). METHODS: With Coptidis Rhizoma (C. Rhizoma) as a case study, the Q-marker of ECI based on the integration of bioeffect and active constituents was developed. According to the efficacies of C. Rhizoma, we investigated its antibacterial and antineoplastic effects by microcalorimetry and MTT assay, respectively. High performance liquid chromatography was performed to determine the active constituents of C. Rhizoma extract simultaneously. ECIS of inhibition on Shigella dysenteriae (S. dysenteriae) and ECIH of inhibition on HepG2 cells were established by multi-indicator synthetic evaluation method. The organoleptic evaluation scores of C. Rhizoma samples were given by Delphi method. RESULTS: The correlation analysis showed that ECIS and ECIH were significantly correlated with the inhibiting effects of C. Rhizoma extract on the growth of S. dysenteriae (P < 0.01) and proliferation of HepG2 cells (P < 0.01), respectively. Moreover, ECI showed a good ability to distinguish and predict the bioeffect-based quality grade, whereas the organoleptic evaluation and chemical analysis failed to achieve it. Plus, some samples with lower ECIS showed higher ECIH and vice versa. CONCLUSIONS: The Q-marker of ECI is useful to associate different pharmacologic effects of C. Rhizoma containing multiple active constituents, which is beneficial for the improvement of quality standard of the CHM in an integrated, convenient, and differentiated way.

Unveiling Active Constituents and Potential Targets Related to the Hematinic Effect of Steamed Panax notoginseng Using Network Pharmacology Coupled With Multivariate Data Analyses.

Steamed Panax notoginseng (SPN) has been used as a tonic to improve the blood deficiency syndrome (BDS) in the theory of traditional Chinese medicine. Here, we aim to unveil active constituents and potential targets related to the hematinic effect of SPN, which has not been answered before. In the study a constituent-target-disease network was constructed by combining the SPN-specific and anemia-specific target proteins with protein-protein interactions. And the network pharmacology was used to screen out the underlying targets and mechanisms of SPN treating anemia. Also, the multivariate data analyses were performed for the double screening. According to the results, 11 targets related to chemical constituents of SPN were found to be closely associated with the hematinic effect of SPN. Among them, the direct target protein of mitochondrial ferrochelatase (FECH) had the major role through the metabolic pathway. Meanwhile, Rk3 and 20(S)-Rg3 were predicted to be major constituents related to the hematinic effect of SPN by both multivariate data analyses and network pharmacology. And it was been validated by the pharmacologic tests that Rk3 and 20(S)-Rg3 could significantly increase the levels of blood routine parameters, FECH and its downstream protein of heme in mice with BDS. The study provides evidences for the mechanism understanding and drug development of SPN for the treatment of anemia.

Uncovering Active Constituents Responsible for Different Activities of Raw and Steamed Panax notoginseng Roots.

Although Panax notoginseng (PN) roots in raw and steamed forms were historically supposed to be different in the efficacies, the raw materials and steamed ones were often undifferentiated in the use and market circulation, which might bring unstable curative effects or even adverse reactions. To uncover chemical constituents responsible to different activities of raw and steamed PN, chemometrics analyses including partial least squares regression (PLSR) and multi-linear regression analysis (MLRA) were used to establish the relationships between the chromatographic fingerprints and activities of PN samples. Chemical fingerprints of PN were determined by HPLC. Anticoagulant and antioxidant activities were evaluated by the thromboplastin inhibiting test and hydroxyl radical scavenging assay, respectively. Results showed that there was a significant difference in the chemical composition between raw and steamed PN, which could be discriminated by principle component analysis according to different steaming temperatures. Compared with the steamed PN, raw PN exhibited stronger anticoagulation and weaker antioxidation. By chemometrics analyses, notoginsenoside R1, ginsenosides Rg1, Re, Rb1, and Rd were found to be the major active constituents of raw PN, whereas ginsenosides Rh1, Rk3, Rh4, and 20(R)-Rg3 had the key role in the activities of steamed PN, which could be used as new markers for the quality control (QC) of steamed PN.