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.

Fingerprint and multi-index content determination of ethyl acetate extract of Sedum emarginatum Migo.

Sedum emarginatum Migo (Aoyejingtian) is a perennial succulent herb of the sedum genus in the family Crassulaceae, which has the fountion of treating furuncle, swelling and haematemesis, hematochezia, menorrhagia and hepatitis. Preliminary studies of our research group had showed that the ethyl acetate extract of Sedum emarginatum Migo could inhibit the proliferation of liver cancer HepG2 cells. The establishment of a reasonable and feasible quality evaluation method for the effective parts of Sedum emarginatum Migo can provide a scientific basis for the further development and utilization of Sedum emarginatum Migo. In this study, a multi-wavelength conversion method was used to establish high-performance liquid chromatography (HPLC) fingerprints of the ethyl acetate extract of Sedum emarginatum Migo, and the method was also used to simultaneously determine the gallic acid, protocatechuic acid, caffeic acid, and ferulic acid, isoquercitrin and luteolin in the ethyl acetate extract of Sedum emarginatum Migo. The similarity of the fingerprints of the ethyl acetate extract of Sedum emarginatum Migo from different origins and the content of 6 components were compared. The established method was simple, accurate, table and reliable, which could provide a fast, accurate and reliable method for comprehensive evaluation of the quality of Sedum emarginatum Migo.