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Objective To investigate the effect of Shuganning injection (SGN) in alleviating drug-induced cholestasis and the possible mechanisms involved. Methods The liver of Sprague-Dawley rats was decellularized to prepare collagen scaffolds, and then the scaffolds were recellularized with human HepG2 cells to obtain the tissue-engineered liver (normal control group). The tissue-engineered liver was perfused with 10 μmol/L chlorpromazine (CPZ) and bile salt mixture to establish a model of drug-induced cholestasis (CPZ group), and the model was further treated with Shuganning injection (10 3 -fold dilution) as the injury protection group (SGN+CPZ group). The markers for hepatocellular injury [alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP)] and the antioxidant and oxidative stress markers [glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS)] were measured for all groups, and the normal control group, the CPZ group, and the SGN+CPZ group were compared in terms of the mRNA and protein expression levels of the enzymes associated with liver bile salt metabolism and the enzymes associated with hepatic cholestasis. HE staining was performed to observe liver pathology. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the CPZ group, the SGN+CPZ group had significant reductions in the markers for hepatocellular injury ALT, AST, LDH, and ALP (all P < 0.000 1), significant increases in the oxidative stress markers GSH and SOD ( P < 0.000 1 and P < 0.001), and significant reductions in the markers MDA and ROS ( P < 0.000 1 and P < 0.001). Compared with the CPZ group, the SGN+CPZ group had significant reductions in the mRNA expression levels of cholesterol 7α-hydroxylase (CYP7A1) and sterol 12α-hydroxylase (CPY8B1) in hepatocytes (all P < 0.001) and significant increases in the mRNA expression levels of farnesoid X receptor (FXR), small heterodimeric partner (SHP), bile salt export pump (BSEP), and multidrug resistance-associated protein 2 (MRP2) ( P < 0.000 1, P < 0.01, P < 0.000 1, and P < 0.000 1). HE staining showed that compared with the CPZ group, the SGN+CPZ group had a significant reduction in hepatocyte injury and a significant increase in the number of cells. Conclusion Shuganning injection can alleviate drug-induced cholestatic liver injury caused by chlorpromazine, and it exerts a protective effect by activating FXR in hepatocytes and increasing the expression of SHP to regulate bile salt balance. It also inhibits CYP7A1 and CYP8B1 to reduce the synthesis of hydrophobic bile acids and upregulates the expression of BSEP and MRP2 to promote the excretion of bile salts.
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This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.
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Animals , Rats , Chlorogenic Acid , Chromatography, High Pressure Liquid , Chromatography, Liquid , Plasma , Rats, Sprague-Dawley , Reproducibility of Results , Tandem Mass SpectrometryABSTRACT
Objective To investigate the clinical liver function protective effect of Shuganning injection for treatment of patients with drug poisoning.Methods One hundred and forty patients with drug poisoning consistent with thecriteria of enrollment into the study were admitted into the Department of Emergency Medicine in the First Hospital of Jilin University-the Eastern Division from January 2015 to August 2016, and they were randomly divided into observation group and control group by the computer generated random numbers, 70 cases in each group. After admission, all the cases were treated with routine treatment including detoxification, removal of toxin, organ protection, symptomatic and supporting treatment, etc. Based on the routine treatment, additionally, Shuganning injection 20 mL+ 10% glucose injection 250 mL, intravenous drip slowly, once a day was given in the observation group; the therapeutic course was 14 days in both groups. The changes of serum glutamic transaminase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) levels were determined before and 7 and 14 days after treatment respectively to evaluate the situation of liver function in the two groups.Results Before treatment, there were no statistical significant differences in the levels of serum ALT, AST and TBil in the two groups (allP > 0.05); with the prolongation of treatment, the above-mentioned indexes of the two groups were gradually reduced, on the 14th day after treatment, they reached the lowest levels, and the degree of decrease in level on the 14th day in the observation group was more significant than that in the control group [ALT (U/L): 32.6±10.8 vs. 98.3±34.5, AST (U/L): 39.4±14.3 vs. 138.5±25.6, TBil (μmol/L): 4.8±1.7 vs. 13.2±2.3, allP < 0.05].Conclusion Shuganning injection has a protective effect on liver function in patients with drug poisoning.
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OBJECTIVE:To investigate the preventive and therapeutic effect of Shuganning injection on alcoholic liver fibrosis (ALF) in model rats,and provide experimental basis for its clinical application for alcoholic liver disease. METHODS:50 rats were enrolled and intraperitoneally given mixed liquid of 60% alcohol-corn oil-pyrazole to reduce ALF model. Another 10 rats were enrolled and intraperitoneally given normal saline,as normal control group. After 16 weeks,survived model rats(n=40)were ran-domly divided into model group,positive control group(Anluo huaxian pill 0.75 g/kg,ig),Shuganning injection high-dose,medi-um-dose,low-dose groups(4.8,2.4,1.2 mL/kg,ip),8 in each group. Normal control group and model group were intraperitone-ally injected equal volume of normal saline (5 mL/kg),administration groups were given relevant medicines,once a day,for 8 weeks;and modeling was contiuously conducted at the same time. After administration,body mass of rats was weighed,and the levels of liver function indexes [aspartate aminotransferase(AST),alanine aminotransferase(ALT)] and liver fibrosis indexes [hyal-uronic acid(HA),laminin(LN),type Ⅲ procollagen(PⅢNP),type Ⅳ collagen(Ⅳ-C)] in serum of rats were detected. Liver index of rats was determined and pathological changes of liver tissue were observed. RESULTS:Compared with normal control group,body mass of rats in model group was significantly decreased(P<0.05);liver index,and liver function index,liver fibro-sis index levels in serum were significantly increased (P<0.05 or P<0.01). Liver tissue showed steatosis,hepatocyte vacuoliza-tion,a large number of fibrous tissue deposition around portal areas and other pathological changes. Compared with model group,above-mentioned changes were improved significantly in administration groups (P<0.05 or P<0.01). CONCLUSIONS:Shugan-ning injection can obviously improve liver tissue damage of model rats with ALF,showing certain preventive and therapeutic effect on alcoholic liver disease.
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OBJECTIVE:To observe the protective effect of Shuganning injection against liver injury induced by cisplatin in mice. METHODS:100 mice were randomized into normal control group (normal saline),model group,Shuganning injection high-dose,medium-dose and low-dose groups [41.5,20.8,10.4 g(crude drug)/kg] with 20 mice in each group. Except normal con-trol group,other groups were given cisplatin 3 mg/kg,ip,on Monday and Thursday to induce cisplatin toxicosis model;at the same time,they were given relevant medicines from Monday to Friday by intraperitoneal injection,once a day for consecutive 10 weeks. The activities of ALT and AST,the contents of albumin,total protein and globin in serum,total protein/globin(A/G)were determined,and hepatic index was calculated and hepatic histomorphology was observed. RESULTS:Compared with normal con-trol group,the activity of AST,the contents of ALB and globin in serum and A/G all increased in model group (P<0.01),un-clear hepatic cords range,hepatocellular albuminoid degeneration and endochylema puffing were common and obvious acidophilic change was found in hepatic tissue. Compared with model group,the serum content of ALB decreased in Shuganning injection low-dose and high-dose groups,while globin content and A/G increased;and the activity of AST in serum decreased in Shuganning high-dose group(P<0.05 or P<0.01),and pathological state of hepatic tissue was improved. CONCLUSIONS:Shuganning injec-tion possesses a protective effect on cisplatin-induced liver injury in mice.