Marein reduces lipid levels via modulating the PI3K/AKT/mTOR pathway to induce lipophagy.

ETHNOPHARMACOLOGICAL RELEVANCE: The capitulum of Coreopsis tinctoria Nutt. (CT, Xue-Ju in Chinese) is a precious medicine in Xinjiang Uygur Autonomous region of China. The Coreopsis tinctoria Nutt. is used to prevent and treat dyslipidemia, coronary heart disease, etc. Recent studies have shown that its extract has a pharmacological effect on hyperlipidemia and hyperglycemia. AIM OF THE STUDY: The study aimed to systematically evaluate the lipid-lowering activity of CT through a mice model of hyperlipidemia and a human hepatoma G2 (HepG2) cells model of lipid accumulation, and to investigate its main active components and mechanism. MATERIALS AND METHODS: Biochemical analysis of blood/liver lipids and liver histopathology were used to evaluate the effect of the aqueous extract of Coreopsis tinctoria Nutt. (AECT) on hyperlipidemia mice. High-performance liquid chromatography (HPLC) analysis was used to identify the main components in the AECT. Oil red O staining, immunofluorescence, western blotting, and determination of the total cholesterol (TC), total triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were used to further study the effect and potential mechanism of the AECT main components on sodium oleate-induced lipid accumulation in HepG2 cells. RESULTS: We confirmed the lipid-lowering activity of the aqueous extract and further identified flavonoids as its main components. Among them, five Coreopsis tinctoria Nutt. flavonoids mixture (FM) significantly reduced lipid droplet area, lipid content, TC, TG, and LDL-C levels, and elevated HDL-C levels in HepG2 cells induced by sodium oleate. Furthermore, they increased lipophagy in HepG2 lipid-accumulating cells, while decreasing the ratio of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR. Most importantly, marein may be a key component. CONCLUSIONS: Our study demonstrated that AECT, with flavonoids as the main component, can improve diet-induced hyperlipidemia in obese mice. Among the main five flavonoids, marein plays a key role in promoting lipophagy by regulating the PI3K/AKT/mTOR pathway, resulting in a lipid-lowering effect.

[Botanical origin identification of Rhei Radix et Rhizoma based on color digitalization].

The color characteristic information of Rhei Radix et Rhizoma powder was obtained by spectrophotometer, the feasibility of rapid identification of Rhei Radix et Rhizoma origin based on chromaticity value was studied by statistical analysis. The results of rank correlation analysis showed that a~*(P<0.01), b~*(P<0.01) had significantly correlation with the origin of medicinal herbs, which could be used as two important parameters to distinguish the origin of Rhei Radix et Rhizoma, the larger the a~* value, the more red the powder color,and the greater the b~* value, the more yellow the powder color. Meanwhile, through Fisher discriminant analysis, the linear discriminant functions of different genus Rhei Radix et Rhizoma were established, which was Rheum tanguticum=40.666a~*+0.019b~*-213.303, Rh. palmatum=34.121a~*+0.061b~*-151.770, Rh. officinale=28.071a~*+0.113b~*-104.604 3, the coincidence rate of cross-validation was over 95%, among them, the discriminant rate of Rh. tanguticum and Rh. officinale reached 100%;In addition, using the percentile method to analyze the 90% reference value range of three different origin of Rhei Radix et Rhizoma, as a result, Rh. tanguticum a~*(10.236 5-10.604 7), b~*(32.294 8-34.841 7); Rh. palmatum a~*(8.602 7-8.770 0), b~*(27.534 8-28.968 6), and Rh. officinale a~*(6.825 7-7.464 3),b~*(21.001 6-27.716 4). According to this study, rank correlation analysis and Fisher discriminant analysis are feasible to distinguish the base of Rhei Radix et Rhizoma in a certain range, and provide some theoretical basis for the identification of Rhei Radix et Rhizoma. It also provides a new method and idea for the identification of other multi-base Chinese medicine.

[Grey correlation analysis and path analysis between isoflavones content in Astragali Radix and climate factors].

In this study, HPLC was used to determine the content of the four isoflavones of Astragalus membranceus var. mongholicus from different regions(calycosin-7-glucoside, ononin, calycosin and formononetin), and gray correlation analysis and path analysis were used to explore the influence of climate factors on the content of isoflavone components in A. membranceus var. mongholicus. The results showed that there were significant differences in the content of the four isoflavones in different areas(P<0.05); grey correlation analysis showed that the highest temperature in July, the lowest temperature in January and the daily average temperature had a greater impact on the content of flavonoid glycosides, meanwhile precipitation and relative humidity were the more important factors for the accumulation of flavonoid aglycones. According to the general analysis, the direct positive effects of the lowest temperature in January and altitude on the contents of four isoflavones in A. membranceus var. mongholicus were significant. High altitude and extreme temperature conditions might be more adverse to the formation and accumulation of isoflavone components. Therefore, the religions of A. membranceus var. mongholicus with high contents of isoflavones should be chosen the low altitude region with higher minimum temperature in January. This study provides a reference basis for the quality evaluation of A. membranceus var. mongholicus, and basic data for the selection of suitable habitat, construction of planting standards and directional cultivation of medicinal materials in A. membranceus var. mongholicus.

[Analysis of flavonoids in Coreopsis tinctoria by integrating 2D-TLC and HPLC-IT-TOF-MS].

The qualitative analysis of flavonoids in Coreopsis tinctoria was carried out by a combination of 2 D-TLC and HPLC-IT-TOF-MS. The separation was conducted on 2 D-TLC and a Phenomenex Kinetex Evo C_(18) column(2.1 mm×100 mm, 2.6 µm) with methanol-0.05% aqueous formic acid by gradient elution. Electrospray ionization-(ESI) source was applied and operated in both positive and negative ionization modes. Eighteen flavonoids including three flavonoids, one flavonol, nine flavonones, one flavanonol and four chalcones, were putatively identified from the flavone-enriched fraction of C. tinctoria. 2 D-TLC could separate the flavonoids from C. tinctoria. HPLC-IT-TOF-MS was able to quickly and accurately analyze the flavonoids in C. tinctoria. The results would provide experimental information for the efficacy material basis clarification of C. tinctoria.