Study on the Fingerprint Spectrum and the Spectrum-Effect Relationship of Analgesic and Anti-Inflammatory Effects of the Aqueous Extract from Dalbergia hancai Benth.

Dalbergia hancai Benth. (D. hancai) is one of the most frequently utilized traditional Chinese medicine in Zhuang medicine. Simultaneously, it has been included in the "Quality Standard of Zhuang medicine in Guangxi Zhuang Autonomous Region (Vol. 2)" and possessed outstanding pharmacological effects. However, the pharmacodynamic material basis of D. hancai still remains unclear. In this study, the high-performance liquid chromatography (HPLC) method had been employed to establish the fingerprint of 10 batches of aqueous extract of D. hancai originated from different parts of China. At the same time, similarity evaluation, cluster analysis, and principal component analysis (PCA) had also been conducted to evaluate the common peaks. The acetic acid-induced writhing in mice had been employed as an analgesic model, and the carrageenan-induced toe swelling in mice was utilized as an anti-inflammatory model for pharmacodynamic experiments. The gray relational analysis (GRA) and partial least squares regression (PLSR) were applied to correlate the fingerprint and pharmacodynamic data to thoroughly examine its spectrum-effect relationship, whereby its analgesic and anti-inflammatory material basis had been comprehensively explored. The results revealed that the HPLC fingerprint of the aqueous extract of D. hancai had successfully identified 12 common peaks whereby two of which were further identified as protocatechuic acid and vitexin. Subsequently, through the analysis of GRA and PLSR, the chromatographic peaks that possess a critical correlation degree with the analgesic and anti-inflammatory effects of D. hancai had also been successfully discovered. Ultimately, the analgesic and anti-inflammatory effects of the 10 batches of D. hancai aqueous extract had been conclusively proved, and it was evidently indicated that these effects were attributable to the synergistic interactions between various components. Therefore, this study aims to serve as an effective analytical method for screening and predicting the effective substances of traditional Chinese medicine on the basis of the spectrum-effect relationship.

Study on the Spectrum-Effect Correlation of Anti-Inflammatory Active Extract of Sauropus spatulifolius Beille.

Sauropus spatulifolius Beille (S. spatulifolius) is a commonly used medicine of the Bourau and Yao nationalities. However, the composition of S. spatulifolius is complex, and simple chemical fingerprints cannot accurately evaluate the relationship between its composition and efficacy. In this study, high-performance liquid chromatography (HPLC) method was used to establish the fingerprint of the ethyl acetate extract of S. spatulifolius. Based on the evaluation of the similarity of chromatographic fingerprints of traditional Chinese medicine, combined with cluster analysis and principal component analysis (PCA), the common peaks of fingerprints were evaluated. The anti-inflammatory effect data were extracted through the dimethylbenzene-induced ear-swelling model in mice. The gray relational analysis (GRA) combined with partial least squares regression (PLSR) was used to study the spectrum-effect correlation of S. spatulifolius. As a result, the HPLC fingerprint of the ethyl acetate extract of S. spatulifolius was established, and 18 common peaks were identified. Except for S6, the other similarities are all above 0.915. The reference substance control method was used to identify two absorption peaks, namely, protocatechuic acid and caffeic acid. The cluster analysis results showed that 10 samples from different origins were grouped into four categories, which was consistent with the PCA results. Ethyl acetate extract of 10 batches of S. spatulifolius could significantly inhibit the ear swelling of mice (P < 0.01). Through GRA, the order of the contribution of each chemical component to the anti-inflammatory efficacy was obtained. The results of PLSR showed that the VIP values of peaks 3, 4, and 12 were greater than 1 and were positively correlated with anti-inflammatory activity. In this study, the HPLC fingerprint of the ethyl acetate extract of S. spatulifolius was established. Through the study of the spectrum-effect correlation, the anti-inflammatory active substance of the ethyl acetate extract of S. spatulifolius was obtained. The anti-inflammatory effect of S. spatulifolius was the result of the joint action of multiple ingredients. This research helps to quickly and accurately discover the active ingredient groups of traditional Chinese medicine and provides new ideas and methods for studying the effective substances of traditional Chinese medicine.

Promotion of HepG2 cell apoptosis by Sedum emarginatum Migo and the mechanism of action.

BACKGROUND: Sedum emarginatum Migo(S. emarginatum) has anti-tumor and anti-oxidant effects. This study aimed to screen the extractions of S. emarginatum against liver cancer in vitro and explore its anti-liver cancer mechanism. METHODS: The CCK-8(Cell Counting Kit-8) method was used to detect the inhibitory effect of different extracts of S. emarginatum on the proliferation of liver cancer HepG2 cells. The morphological changes of the cells after administration were observed with microscopy, cell apoptosis was detected by flow cytometry, and the expression of Bax, Bcl-2 and Caspase-3 mRNA in the cells were detected by RT-PCR (Reverse Transcription-Polymerase Chain Reaction) to explore the mechanism of action. RESULTS: CCK-8 method test results showed that among the different extracts of S. emarginatum, the ethyl acetate extract(1000 µg/ml, 2000 µg/ml, 2500 µg/ml, 3000 µg/ml) and n-butanol extract(1000 µg/ml, 2000 µg/ml, 2500 µg/ml, 3000 µg/ml) have the strongest inhibitory effect on the proliferation of HepG2 cells. In these 4 concentrations, the inhibitory effect increased as the concentration increased. The IC50 of the ethyl acetate extract on HepG2 cells was less than that of the n-butanol extract, so the ethyl acetate extract has a better proliferation inhibitory effect on HepG2 cells than the n-butanol extract, followed by the 70% ethanol extract(3000 µg/ml) and the water extract(3000 µg/ml), petroleum ether extract was the weakest. The results of microscopy showed that ethyl acetate extract caused hepatocarcinoma HepG2 cell morphology changed, cell density decreased, and suspension cells increased. Moreover, the results of flow cytometry showed that the ethyl acetate extract of S. emarginatum could induce HepG2 cell apoptosis at the concentrations of 2500µg/ml and 3000µg/ml. RT-PCR results showed that the expression of Bax mRNA was up-regulate by the middle(2500 µg/ml) and high(3000 µg/ml) dose groups of ethyl acetate extract. The expression of Caspase-3 mRNA was up-regulated by the low(2000 µg/ml), medium(2500 µg/ml) and high(3000 µg/ml) dose groups of ethyl acetate extract. The expression of Bcl-2 mRNA was down-regulated by the high(3000 µg/ml) dose group of ethyl acetate extract. CONCLUSION: The ethyl acetate extract of S. emarginatum has the best effect on human liver cancer HepG2 cells. Its anti-hepatocellular mechanism may be related to affect the expression of apoptosis genes (Bax, Bcl-2 and Caspase-3mRNA) and promote the apoptosis of liver cancer cells. It provided a reference for the research and development of drugs for the treatment of liver cancer.

[Macroscopic and microscopic identification of Zhuang medicine Tetrastigma planicaule].

OBJECTIVE: To study the macroscopic and microscopic characteristics of Zhuang medicine Tetrastigma planicaule. METHODS: Macroscopic and microscopic identification were studied. RESULTS: The microscopic characteristics were significant. The root had more than 10 layers of cork cells arranging in line. The starch grain and calcium oxalate cluster crystals distributed in the phloem parenchymas, and mucilage cells scattered in the cortex of the stem. There were 8 vascular bundles in the vein of leaves. CONCLUSION: This study provides a scientific basis for the identification and utilizing of Tetrastigma planicaule.

[Studies on the chemical constituents of the petroleum ether portion of Nervilia fordii].

OBJECTIVE: To study the chemical constituents of the petroleum ether portion of Nervilia fordii. METHODS: The constituents were separated and purified by using column chromatography with silica gel. These compounds were identified by their physical and spectral data. RESULTS: Six compounds were isolated and identified as Cyclohomonervilol (I), Octacosanoicacid (II), Stigmasterol (III), Cyclohomonervilol-(E)-p-hydroxy cinnamate (IV), 24(R/alpha)-dihydrocycloeucalenol-(E)-p-hydroxy cinnamate (V), Docosanoic acid (VI). CONCLUSION: Compounds I-VI are isolated from this plant for the first time.

[Study on fingerprints of the stems of Actinidia deliciosa by HPLC].

OBJECTIVE: To establish the fingerprint spectrum of the stems of Actinidia deliciosa by HPLC. METHODS: HPLC fingerprint analysis of the stems of Actinidia deliciosa was developed and the thermo Hypersil BDS C18 column (250 mm x 4.6 mm, 5 microm) was used. The mobile phase consisted of Acetonitril-0.1% phosphoric acid with gradient elution. The column temperature was 20 degrees C, the detective wavelength was 296 nm, the flow rate was 1.0 mL/min, and the sample injection was 20 microL. RESULTS: Fingerprint spectrum of the stems of Actinidia deliciosa was established, and 16 samples of different origin Actinidia deliciosa were detected. Twenty-eight peaks in chromatogram were common. There was a high similarity and each chromatographic peak was obtained with good separation correlation according to the technical requirements of fingerprint of Chinese traditional medicine. CONCLUSION: This method is accurate, reliable and provides a scientific basis for controlling the quality of the stems of Actinidia deliciosa.

[Research on chemical constituents from stem of Gymnema sylvestre].

OBJECTIVE: To study on the chemical constituents from the stem of Gymnema sylvestre. METHODS: The constituents were extracted by percolation with ethanol. Then the extract was separated by systemic solvent separation methods. The part of n-butanol extract was isolated and purified by macroporous adsorptive resins, silica gel column chromatography, sephadex gel column chromatography and recrystallization. The isolated compounds were identified by spectrum methods. RESULTS: Eight compounds were isolated and identified as fallows: Conduritol A(I), 1-Heptadecanol(II), Stigmasterol glucoside(III), 1-Quercitol(IV), 1-Octadecanol(V), Potassium nitrate(VI), Lupeol cinnamate(VII), Stigmasterol(VIII). CONCLUSION: Chemical compounds II, III, V, VII are firstly obtained from this plant.

[Analysis of the volatile oils chemical constituents of roots of Actinidia deliciosa].

OBJECTIVE: To analyze the volatile oils chemical constituents of roots of Actinidia deliciosa. METHODS: The volatile oils fraction of roots of Actinidia deliciosa. were extracted by water vapor distilling, and then the constituents were separated and identified, by GC-MS. RESULTS: 16 compounds were identified, accounting for 89.37% of all quantity. CONCLUSION: The principal volatile oils chemical constituents are Phenol, 2,4-bis(1,1-dimethylethl)-; 2-Propenoic acid, 3-(4-methox yphenyl)-, ethyl ester; 9-Octadecenoic acid (Z)-, methyl ester; Cyclotetrasiloxane, octamethyl-.

[Studies on chemical constituents in n-butyl alcohol extract of roots of Actinidia deliciosa in Guangxi].

OBJECTIVE: To study the chemical constituents of n-butyl alcohol extract in the roots of Actinidia deliciosa in Guangxi. METHOD: The constituents were separated with various chromatographic techniques and their structures were elucidated by means of physicochemical properties and the analysis of their spectral data. RESULT: Six compounds were isolated and identified as eriantic acid B (1), 2alpha, 3beta, 24-trihydroxyursa-12-en-28-oic acid (2), 2alpha, 3alpha, 24-trihydroxyursa-12-en-28-oic acid (3), 2alpha, 3alpha, 23-tri-hydroxyursa-12, 20 (30)-dien-28-oic acid (4), 2alpha, 3alpha, 24-trihydroxyursa-12, 20 (30)-dien-28-oic acid (5), n-butyl-O-beta-D-fruto-pyranoside (6). CONCLUSION: Compounds 1-4, 6 were obtained from this plant for the first time. Compound 6 was obtained from the genus Actinidia for the first time.

[Experimental [corrected] study of hypoglycemic activity of conduritol A of stems of Gymnema sylvestre].

OBJECTIVE: To investigates the mechanism of hypooglycemic effect of conduritol A of stems of Gymnema sylvestre. METHOD: Fourteen days later after administration, observation is taken on the change of these mice and rats weight, the FBG, TG, CHO, SOD, MDA, INS, TNF in serum were also detected with enzymology method and Radioimmuoassay method. Take the liver to determine the disposal of glucose. Take the pancreas to do the HE and immunohistochemistrial staining, and show pancreas islet beta-cell. Calulate thymus, pancreas, splenica index. RESULT: Compared with diabetic model mice, high and middosage of conduritol A could remarkably reduce fasted blood sugar in diabetic rats induced by alloxan (P < 0.01). Significantly increase the level of serum insulin (P < 0.05). Activity of SOD was obviously increased, and amount of MDA was obviously decreased (P < 0.05). The amount of conduritol A disposal of glucose was obviously increased (P < 0.05). Significantly increase thymus, pancreas, splencia index (P < 0.01 or 0.05); inhibited the atrophy of thymus, pancreas, splencias of the diabetic rats induced by alloxan. Compared with diabetic model group, cell structure and form of conduritol A had been some way improved. The immunohistochemistry results showed that beta-cells numbers of pancreas in each conduritol A group were more than those in the model group. CONCLUSION: Conduritol A could have an effect on regulating the metabolism of blood lipid, free-radical scavenging, enhancing the antioxidant ability, potentiating immune function. Promoting synthesis of hepatic to decrease fasted blood suger.

[Studies on the chemical constituents of the ethyl acetate portion of Nervilia fordii].

OBJECTIVE: To study the chemical constituents of the ethyl acetate portion in the herb of Nervilia fordii from guangxi. METHODS: The constituents were separated and purified by using column chromatography with silica gel. These compounds were identified by their physical and spectral data. RESULTS: Five compounds were isolated and identified as norleucine (crystal I), 24 (S/beta)-dihydrocycloeucalenol-(E)-p-hydroxy cinnamate (crystal II) , rhamnocitrin (crystal III), rhamnazin (crystal IV), daucosterol (crystal V). CONCLUSION: Compounds I , II, III, IV, V were isolated from this plant for the first time.

[Study on anti-tumor effect of extractions from roots of Actinidia deliciosa].

OBJECTIVE: To study the anti-tumor effect of ADEE and ADBE in mice. METHODS: The models of S180 entity tumor mice and H22 bearing tumor mice were established to observe the effect of ADEE and ADBE on inhibiting S180 entity tumor growth and prolonging life time of H22-mice, and the effect on spleen and thymus index to S180 entity tumor. RESULTS: Both ADEE and ADBE had ohvious anti-tumor effects to S180-mice (P < 0.05, P < 0.01) and could prolong H22-mice life (P < 0.05, P < 0.01). In certain degree, ADEE and ADBE could improve the immune function of bearing tumor mice. CONCLUSION: ADEE and ADBE have obvious anti-tumor effect.

[Study on anticancer effect in vivo of active fraction from Nervilia fordii].

OBJECTIVE: To determine the active fraction with anticancer effect in vivo from Nervilia fordii. METHODS: The effective petroleum ether extract and ethyl acetate extract parts preliminary were selected in vitro, then anticancer experiments in vivo were done by S180-mice and H22-mice models. RESULTS: Petroleum ether extract and ethyl acetate extract parts both had obvious anticancer effects to S180-mice and H22-mice, and could prolong H22-mice life. Meanwhile, they could improve the immunoloregulation of mice. CONCLUSION: It is the first time that the petroleum ether extract and ethyl acetate extract of Nervilia foadii are proved to be the effective anticancer fractions in vivo. On this basis, the further studies are needed on active principles or principle group with anticancer effect and the characteristics of this effect in Nervilia foadii.