ABSTRACT
The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.
Subject(s)
Animals , Cattle , Mice , Apoptosis , Cells, Cultured , Fatty Liver , Fructose , Gallstones , Chemistry , Hepatocytes , Cell Biology , Metabolism , Lipid Metabolism , Lipid Peroxidation , Liver , Medicine, Chinese Traditional , Reactive Oxygen Species , Metabolism , Serum , Chemistry , Sterol Regulatory Element Binding Protein 1 , Metabolism , TriglyceridesABSTRACT
Objective To develop an HPLC-MS/MS method for the simultaneous determination of 26 bile acids in Calculus Bovis Sativus (CBS) and Calculus Bovis (CB). Methods Chromatographic separation was performed on a Symmetry C18 column (150 mm × 2.1 mm, 3.5 μm) with linear gradient elution of water (0.1% formic acid + 10 mmol/L ammonium acetate) and methanol (0.1% formic acid + 10 mmol/L ammonium acetate) at a flow rate of 0.15 mL/min, and the injection volume was 10 μL. Switching electrospray ion source polarity in negative mode. The ion spray voltage was set at - 4 500 V, the turbo spray temperature was maintained at 350 ℃, and multiple-reaction monitoring (MRM) was employed. Results The 26 bile aicds showed good linearity within the linear ranges (r ≥ 0.991 4). The average recoveries of the bile acids ranged from 98.2% to 102.3%. The precisions (RSD) for the investigated components were less than 0.95%. There are some differences in variety and chemical components between CBS and CB. Conclusion The proposed method is reliable and has high sensitivity and good specificity, which allows the determination of various bile acids in CB.
ABSTRACT
Objective To observe the effect of saline gauze tamponade of calculus bovis sativus on the level of IL-6 and IL-8 in the tissue of patients with perianal abscess after operation,and to explore its possible mechanism of action in promoting wound healing.Methods Patients with 84 cases of anal abscess surgery,were randomly divided into treatment group(n=42)and control group(n=42).Treatment group calculus bovis sativus research pressed into powder,take 0.10~0.15g evenly scattered in the saline gauze on the wound packing,the control group was treated with physiological saline gauze wound.After treatment,3,7 and 10d were treated with local granulation tissue about 0.2cm×0.2cm×0.1cm size,the IL-6 and IL-8 cytokines in local tissues were detected by enzyme-linked immunosorbent assay.Results There was no significant difference in the expression of IL-6 and IL-8 between the two groups 3 days after treatment(P>0.05),IL-6 and IL-8 in the treatment group(7,10d)were significantly lower than that in control group,and the difference was statistically significant(P<0.05).Conclusion Calculus bovis sativus in the treatment group,IL-6 and IL-8 levels were significantly lower than those of the control group,indicating that calculus bovis sativus can reduce local inflammation response,promote healing of wound after perianal abscess operation.
ABSTRACT
Objective To make a qualitative and a quantitative control on the Calculus Bovis in Houzao Niuhuang Powder.Methods Hyodeoxycholic acid and bilirubin in Calculus Bovis were separately identified by TLC,and the cholic acid content was determined by TLCS with the scan wavelength of ?s=380 nm,?R=650 nm.Results TLC can be used for the identification in-vitro cultured Calculus Bovis and artificial Calulus Bovis,the samples containing cultured Calulus Bovis showed the specific spots of bilirubin but no hyodeoxycholic acid spot while the samples containing artifical Calulus Bovis showed the hyodeoxycholic acid spot but no bilirubin spot.TLC-scanning can be used to measure the quantity of cholic acid,with the linear range being 0.2～1.8 ?g,average recovery rate of 99.5 %,and RSD being 3.3 %(n=6),and the results accorded with the criterion of the cultured Calculus Bovis in Chinese pharmacopoeia.Conclusion The methods can effectively control the quality of Calculus Bovis in Houzao Niuhuang Powder.