1.Research on the fingerprint and three active components assay in Shenshuaining granules by HPLC
Xinhua SONG ; Xujiao CHEN ; Fengyi DENG ; Shouhong GAO ; Hui PENG
Journal of Pharmaceutical Practice 2020;38(3):259-263
Objective To establish the fingerprint spectrum and assay three active components (hesperidin, salvianolic acid B and chrysophanol) in Shenshuaining granule by HPLC method. Methods The chromatographic separation was achieved on SunFireTM C18 column with acetonitrile-0.1% formic acid aqueous solution as mobile phase. Gradient elution program was applied with flow rate of 1.0 ml/min, detection wavelength at 254 nm and the column temperature at 25 ℃. The fingerprint spectrum was established and three active components in Shenshuaining granule were assayed. Results There were 22 common peaks on the fingerprints after analyzing chromatograms from 10 batches of Shenshuaining granules. Good fingerprint similarities (≥0.9) between different batches and the control chromatogram were found. This method has great repeatability, stability and precision, which meets all the assay requirements. Conclusion A simple and reliable HPLC method was developed, which is suitable for the fingerprint establishment of Shenshuaining granules. It provides a method for the quality control of Shenshuaining granules.
2.Fingerprint establishment of Huangqin decoction and study on spectrum-effect relationship of its antidermatophytic activity in different phase states
Chengying SHEN ; Wenming WU ; Xiongjun HOU ; Fengyi DENG ; Jie HE ; Jianxin HU
China Pharmacy 2023;34(6):687-692
OBJECTIVE To establish the fingerprint of Huangqin decoction (HQD), to separate the phase states and screen the active phase states of antidermatophytic activity so as to study the spectrum-effect relationship. METHODS HPLC method was adopted using baicalin as reference, the fingerprints of 10 batches of HQD were drawn and the similarity evaluation was carried out using the Similarity Evaluation System of Chromatographic Fingerprint of TCM (2012 edition) to determine the common peak; the phase states of HQD were separated and characterized by high-speed centrifugation and membrane dialysis. The minimum inhibitory concentrations (MIC) of HQD and its different phase states against Trichophyton mentagrophytes were determined simultaneously. Using the peak area of 37 common peaks as independent variable, MIC as dependent variable, Pearson correlation analysis was performed by using SPSS 21.0 software. RESULTS A total of 37 common peaks were obtained in HPLC fingerprints of 10 batches of HQD, with the similarity higher than 0.99. Ten components were identified, such as albiflorin, paeoniflorin, liquiritin apioside, baicalin, melaleuca glycoside A, wogonoside, baicalein, glycyrrhizic acid, wogonin and oroxylin A. HQD was split into 3 phase states, such as precipitation phase (HQD-P), solution phase (HQD-S) and nano phase (HQD-N). The morphology of HQD-P was irregular granular, and the average particle size was 4.670-91.522 μm. The morphology of HQD-S was uniform flakes, and no particle size was detected. HQD-N was spherical in shape and the particle size was (129.0±12.9) nm. MIC values of each phase state of HQD against T. mentagrophytes in different phase states were HQD-N (4.64 mg/mL) <HQD (5.85 mg/mL) <HQD-P (7.37 mg/mL) <HQD-S (12.89 mg/mL) at the same dosage. Pearson correlation analysis showed that the peak area of 25 of the 37 common peaks (including identified components) was significantly negatively correlated with MIC (absolute values of correlation coefficient>0.95 and P<0.05). CONCLUSIONS The chemical composition of 10 batches of HQD is consistent; HQD-N is the active phase state of HQD. Ten components such as paeoniflorin, liquiritin apioside and baicalin may be the main active components of HQD. The antidermatophytic effect of HQD is closely related to its component content and physical phase state.
3.Study on the antifungal activity and mechanism of Huangqin decoction against Trichophyton mentagrophytes
Chengying SHEN ; Zhong LUO ; Pei ZHANG ; Fengyi DENG ; Baode SHEN ; Jianxin HU
China Pharmacy 2024;35(3):311-315
OBJECTIVE To study the antifungal activity of Huangqin decoction (HQD) against Trichophyton mentagrophytes and explore its mechanism. METHODS Minimal inhibitory concentration (MIC), minimal fungicidal concentration (MFC), mycelial length, spore germination rate, biomass and mycelium ultrastructure observation were performed to evaluate the antifungal activity of HQD against T. mentagrophytes. The effects of HQD on the cell wall of T. mentagrophytes were detected through sorbitol protection experiment. By measuring the content of ergosterol and the activities of squalene epoxide (SE) and lanosterol 14α-demethylase (CYP51), the activity of HQD on the cell membrane of T. mentagrophytes was investigated. The effects of HQD on T. mentagrophytes mitochondria were investigated by determining the activities of malate dehydrogenase (MDH), succinate dehydrogenase (SDH), and ATPases (including sodium potassium ATPase, calcium magnesium ATPase, and total ATPase). RESULTS HQD exhibited significant antifungal activity against T. mentagrophytes with MIC of 3.13 mg/mL and MFC of 25 mg/mL. After intervention with HQD, the mycelial length of T. mentagrophytes was significantly shortened (P<0.05); spore germination rate, biomass, the content of ergosterol in the cell membrane, the activities of SE and CYP51 in the cell membrane and MDH, SDH and ATPase in mitochondria were all decreased significantly (P<0.05); cell structure had been ;damaged to a certain extent, but the integrity of the cell wall had not been affected. CONCLUSIONS HQD shows significant antifungal activity against T. mentagrophytes, the mechanism of which may be associated with reducing the 0791- content of ergosterol in the cell membrane and the activities of SE, CYP51, and mitochondria-related enzymes.
4.Protective effect of dulaglutide against sepsis⁃induced acute kidney inj ury in mice
Fengyi Deng ; Yue Wang ; Xingyu Fan ; Hujing Deng ; Xing Zhong ; Yijun Du ; Hong Su ; Tianrong Pan
Acta Universitatis Medicinalis Anhui 2023;58(8):1329-1334
Objective :
To investigate the protective effect of dulaglutide on acute kidney injury (AKI) induced by
lipopolysaccharide (LPS) .
Methods :
Twenty⁃four male C57BL/6 mice were randomly divided into Control group (normal saline) , LPS group (LPS 15 mg/kg) , LPS + Dul group (LPS 15 mg/kg + Dulaglutide 0. 6 mg/kg) and Dul group (Dulaglutide 0. 6 mg/kg) with 6 mice in each group. The drug was administered by intraperitoneal injection. After drug intervention for 24 h , the body weight and kidney weight of mice were recorded , and kidney tissue and serum samples were collected. The pathological changes in kidney tissue were observed by HE staining.
The serum urea nitrogen (BUN) and creatinine (CRE) levels were detected by the kit. The levels of cytokines interleukin (IL⁃6) , tumor necrosis factor (TNF⁃α ) and IL⁃1β in the kidney were detected by qRT⁃PCR. The contents of macrophage marker F4/80 and myeloperoxidase (MPO) in kidney were determined by immunohistochemistry.
Results :
Compared with Control group , mice in LPS group lost weight and increased kidney weight ( P <
0. 001) . Moreover, the levels of BUN and CRE increased (P < 0. 001 , P < 0. 01) . Meanwhile , the mRNA levels of IL⁃6 , IL⁃1β and TNF⁃α increased (P < 0. 05) . There was obvious pathological damage in kidney tissue. In addition , macrophage and neutrophil infiltration increased in LPS group ( P < 0. 001) . Compared with LPS group , mice in LPS + Dul group gained weight and lost kidney weight (P < 0. 05 , P < 0. 001) . Moreover, the levels of BUN and CRE in LPS + Dul group decreased (P < 0. 01) . The renal histological scores were reduced (P < 0. 05) . In addition , the levels of IL⁃6 , IL⁃1β and TNF⁃α in kidney tissue decreased (P < 0. 05 or P < 0. 01) . Moreover, the infiltration of macrophages and neutrophils in kidney was reduced (P < 0. 01) .
Conclusion
Dulaglutide has a protective effect on LPS⁃induced sepsis AKI , which may be related to reduce the expression of inflammatory media⁃ tors and decrease the infiltration of inflammatory cell.