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1.
China Journal of Chinese Materia Medica ; (24): 6075-6081, 2023.
Article in Chinese | WPRIM | ID: wpr-1008806

ABSTRACT

With the continuous exploration of microemulsions as solvents for traditional Chinese medicine extraction, polyoxyethy-lene(35) castor oil(CrEL), a commonly used surfactant, is being utilized by researchers. However, the problem of detecting residues of this surfactant in microemulsion extracts has greatly hampered the further development of microemulsion solvents. Based on the chemical structures of the components in CrEL and the content determination method of castor oil in the 2020 edition of the Chinese Pharmacopoeia(Vol. Ⅳ), this study employed gas chromatography(GC) and single-factor experiments to optimize the preparation method of methyl ricinoleate from CrEL. The conversion coefficient between the two was validated, and the optimal sample preparation method was used to process microemulsion extracts of Zexie Decoction from three batches. The content of methyl ricinoleate generated was determined, and the content of CrEL in the microemulsion extracts of Zexie Decoction was calculated using the above conversion coefficient. The results showed that the optimal preparation method for CrEL was determined. Specifically, 10 mL of 1 mol·L~(-1) KOH-methanol solution was heated at 60 ℃ for 15 min in a water bath. Subsequently, 10 mL of boron trifluoride etherate-methanol(1∶3) solution was heated at 60 ℃ for 15 min in a water bath, followed by extraction with n-hexane twice. CrEL could stably produce 20.84% methyl ricinoleate. According to this conversion coefficient, the average mass concentration of CrEL in the three batches of Zexie Decoction microemulsion extracts was 11.94 mg·mL~(-1), which was not significantly different from the CrEL mass concentration of 11.57 mg·mL~(-1) during microemulsion formulation, indicating that the established content determination method of this study was highly accurate, sensitive, and repeatable. It can be used for subsequent research on microemulsion extracts of Zexie Decoction and provide a reference for quality control of other drug formulations containing CrEL.


Subject(s)
Polyethylene Glycols/chemistry , Castor Oil , Methanol , Surface-Active Agents/chemistry , Solvents , Water/chemistry , Emulsions/chemistry
2.
China Journal of Chinese Materia Medica ; (24): 5851-5862, 2023.
Article in Chinese | WPRIM | ID: wpr-1008783

ABSTRACT

This study investigated the mechanism of Zexie Decoction(ZXD) in promoting white adipose tissue browning/brown adipose tissue activation based on the GLP-1R/cAMP/PKA/CREB pathway. A hyperlipidemia model was induced by a western diet(WD) in mice, and the mice were divided into a control group, a model group(WD), and low-, medium-, and high-dose ZXD groups. An adipogenesis model was induced in 3T3-L1 cells in vitro, and with forskolin(FSK) used as a positive control, low-, medium-, and high-dose ZXD groups were set up. Immunohistochemistry and immunofluorescence results showed that compared with the WD group, ZXD promoted the expression of UCP1 in white and brown adipose tissues, and also upregulated UCP1, CPT1β, PPARα, and other genes in the cells. Western blot analysis showed a dose-dependent increase in the protein expression of PGC-1α, UCP1, and PPARα with ZXD treatment, indicating that ZXD could promote the white adipose tissue browning/brown adipose tissue activation. Hematoxylin-eosin(HE) staining results showed that after ZXD treatment, white and brown adipocytes were significantly reduced in size, and the mRNA expression of ATGL, HSL, MGL, and PLIN1 was significantly upregulated as compared with the results in the WD group. Oil red O staining and biochemical assays indicated that ZXD improved lipid accumulation and promoted lipolysis. Immunohistochemistry and immunofluorescence staining for p-CREB revealed that ZXD reversed the decreased expression of p-CREB caused by WD. In vitro intervention with ZXD increased the protein expression of CREB, p-CREB, and p-PKA substrate, and increased the mRNA level of CREB. ELISA detected an increase in intracellular cAMP concentration with ZXD treatment. Molecular docking analysis showed that multiple active components in Alismatis Rhizoma and Atractylodis Macrocephalae Rhizoma could form stable hydrogen bond interactions with GLP-1R. In conclusion, ZXD promotes white adipose tissue browning/brown adipose tissue activation both in vivo and in vitro, and its mechanism of action may be related to the GLP-1R/cAMP/PKA/CREB pathway.


Subject(s)
Mice , Animals , Adipose Tissue, Brown , Molecular Docking Simulation , PPAR alpha/metabolism , Adipose Tissue, White , RNA, Messenger/metabolism
3.
China Journal of Chinese Materia Medica ; (24): 453-460, 2022.
Article in Chinese | WPRIM | ID: wpr-927989

ABSTRACT

The present study investigated the pharmaceutical effect and underlying mechanism of Zexie Decoction(ZXD) on nonalcoholic fatty liver disease(NAFLD) in vitro and in vivo via the LKB1/AMPK/PGC-1α pathway based on palmitic acid(PA)-induced lipid accumulation model and high-fat diet(HFD)-induced NAFLD model in mice. As revealed by the MTT assay, ZXD had no effect on HepG2 activity, but dose-dependently down-regulated alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the liver cell medium induced by PA, and decreased the plasma levels of ALT and AST, and total cholesterol(TC) and triglyceride(TG) levels in the liver. Nile red staining showed PA-induced intracellular lipid accumulation, significantly increased lipid accumulation of hepatocytes induced by PA, suggesting that the lipid accumulation model in vitro was properly induced. ZXD could effectively improve the lipid accumulation of hepatocytes induced by PA. Oil red O staining also demonstrated that ZXD improved the lipid accumulation in the liver of HFD mice. JC-1 staining for mitochondrial membrane potential indicated that ZXD effectively reversed the decrease in mitochondrial membrane potential caused by hepatocyte injury induced by PA, activated PGC-1α, and up-regulated the expression of its target genes, such as ACADS, CPT-1α, CPT-1β, UCP-1, ACSL-1, and NRF-1. In addition, as revealed by the Western blot and immunohistochemistry, ZXD up-regulated the protein expression levels of LKB1, p-AMPK, p-ACC, and PGC-1α in vivo and in vitro. In conclusion, ZXD can improve NAFLD and its mechanism may be related to the regulation of the LKB1/AMPK/PGC-1α pathway.


Subject(s)
Animals , Mice , AMP-Activated Protein Kinases/metabolism , Alanine Transaminase/metabolism , Diet, High-Fat , Liver/metabolism , Mice, Inbred C57BL , Non-alcoholic Fatty Liver Disease/genetics , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
4.
Chinese Herbal Medicines ; (4): 290-297, 2018.
Article in Chinese | WPRIM | ID: wpr-842119

ABSTRACT

Objective: To investigate the effects of fluidized bed granulation with dextrin on moisture sorption and diffusion of Zexie Decoction granules. Methods: The particle characterization was studied by the particle size, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR). The moisture sorption isotherm, equilibrium moisture content (EMC), and moisture diffusion coefficients were determined by using the saturated salt solution method. Results: The particle size increased from 6.04 µm (powder) to 1201.47 µm (granule). The glass transition temperature of dextrin, Zexie Decoction powder, and granule was 107.13 °C, 94.82 °C, and 126.25 °C. As the increase of temperature, the initial rate of moisture sorption become higher. Furthermore, the initial rate of moisture sorption of Zexie Decoction granules was lower than those of powders and dextrin. The EMC and moisture diffusion coefficients were reduced significantly after granulation (P < 0.01). Critical relative humidity and diffusion activation energy of granules were higher than powders. Conclusion: Results suggested that fluidized bed granulation with dextrin could reduce the hygroscopicity of the Zexie Decoction extract powders and inhibited moisture diffusion, which is mainly related to the microstructure reorganization by fluidized bed granulation and anti-plasticizing effects of dextrin.

5.
Tianjin Medical Journal ; (12): 1193-1196,1253, 2014.
Article in Chinese | WPRIM | ID: wpr-600056

ABSTRACT

Objective To explore the effect of modified Zexie Decoction on renal aquaporin-2 (AQP2) expression in high-salt hypertensive rat. Methods Hypertensive rats model was established by feeding rat with 8%high salt. Rats (n=50) were divided into model group, modern medicine group, traditional Chinese medicine groups of high, medium, low dose, with 10 rats in each group. The other 10 rats were fed with ordinary diet as normal group. Rats in traditional Chinese medi?cine of high, medium, low groups were given Zexie Decoction suspension of 16.2, 10.8 and 5.4 g/(kg·d) respectively;Rats in modern medicine group was given Valsartan hydrochlorothiazide 16.65 mg/(kg·d);the model group and normal group was ad?ministered with equal volume of distilled water. Animals were feed with medications at 1 mL/100 g by gavage for 4 weeks. On the 1st , 4th , 7th , 14th and, 28th day of administration, we measured SBP and collected 24 h urine. We employed immunohis?tochemistry to detect renal AQP-2 protein expression level and RT-PCR to detect renal AQP-2 mRNA transcription level. Results The rank of SBP from high to low is:model group>traditional Chinese medicine medium and low dose groups>traditional Chinese medicine high dose group and western medicine group>normal group. The rank of urine volume from high to low is:Western medicine group and traditional Chinese medicine high dose group>traditional Chinese medicine me?dium and low dose group > normal group, the difference was not statistically significant between traditional Chinese medi?cine medium and low dose group , or between western medicine group and traditional Chinese medicine high dose group. The renal AQP-2 in epithelial cells along the collecting duct wall of rats in model group show brown particles which are darker and wider distributed than those in normal group and traditional Chinese medicine of high, medium, low dose groups. RT-PCR results show that AQP-2 mRNA expression is highest in rats of model group and lowest in rats of traditional Chi?nese medicine high dose group (P<0.05). No statistical significance of AQP mRNA level was found between traditional Chi?nese medicine low group and model group (P < 0.05). Conclusion Modified Zexie Decoction can lower blood pres?sure by inhibiting the expression of AQP-2.

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