The Role Of Shuanghuanglian Freeze-dried Powder For Injection Combined With Cefradine For Injection On HL-77O2

BackgroundsShuanghuanglian freeze-dried powder for injection composed by Forsythia, Honeysuckle, Radix scutellariae. its main components are bacalin, chlorogeni c acid, forsythin, wogonin. it is applied to treat with the diseases caused by the exogenous wind-heat, evil in the lung,such as fever,cough, breathl ess, Cough of yellow sputum,sore throat in TCM theory, and many kinds of a cute upper respiratory tract infection, acute bronchitis, acute tonsillitis, light pneumonia. Cefradine for injection is the first generation cephalos porin antibiotic.it is applied to treat repiratory tract infection,urina ry tract infections, skin soft-tissue infection such as acute pharynitis, tonsillitisotitis, media bronchitis, pneumonia caused by sensitive bacteria. Together they were used in treatment of bacteria combined with virus inf ection, what they caused liver toxicity assessment has not been reported.ObjectiveWe chose hepatocyte culture in vitro as our reseath object, detecting cell viability IC50, the indexes of liver functions and cell morphology, in order to study the effect of ShuangHuanglian freeze-dried powder for injection and with Cefradine for injection on HL-7702.Methods and Results1.Cell toxicity test of Shuanghuanglian freeze-dried powder for injection combined with cefradine for injection on HL-7702.1.1Study on biological characteristics of HL-7702cells HL-7702growth curve drawning, screening for drug concentration. MTT select the appropriate stimulation concentration of Shuanghuanglian freeze-dried powder for injection combined with Cefradine for injection.HL-7702cells were divided into ten group:solvent control group, cefradine (0.046mg/mL), SHL(0.25mg/mL), SHL(0.5mg/mL), SHL(1. Omg/mL), SHL(2. Omg/mL), SHL+cefradine(0.25mg/mL+0.046mg/mL), SHL+cefradine(0.5mg/mL+0.046mg/mL), SHL+c efradine(1. Omg/mL+0.046mg/mL), SHL+cefradine(2.0mg/mL+0.046mg/mL).1.2Detect liver function indicatorDetection of liver function indicator include LDH, AST, ALT, SOD, MDA. compared with solvent control group,the experimental group in dose-response manner, with the increase of dosage, the content of SOD decreased, and the content of ALT, AST, LDH, MDA increased gradually (p<0.05).1.3Detecting the pathological changes of Shuanghuanglian freeze-dried powder for injection combined with Cefradine for injection on HL-7702by HE staining.the result as follows:at low dose, the distance of HL-7702cells increase, the connection of the cells disappeared, the shape of the cells become irregular and a small number of the cells fall off;at high dose, many nuclei have become pyknotic(shrunken and dark), empty bubble formed inside the cell. Shuanghuanglian freeze-dried powder for injection combined with Cefradine for injection aggravate hepatotoxicity.2. The determination of ATP, ADP of Shuanghuanglian freeze-dried powder for injection combined with Cefradine for injection on HL-7702Experiment results show that the content of ADP and ATP decreased gradually as the drug concentration increased.3. Effect on the expression of Shuanghuanglian freeze-dried powder for injection combined with cefradine for injection on HL-7702cells HO-1, COX2Using RT-PCR detect the expression of HO-1,COX2mRNA.their expression increased gradually as the drug concentration increased, compared with solvent control group, SHL+cefradine group is especially remarkable.4. Possible way of Shuanghuanglian freeze-dried powder for injection combined with cefradine for injection on HL-7702.Determination content of Chlorogenic Acid, Forsythin, Baicalin and Wogonin in Shuanghuanglian freeze-dried powder for injection by HPLC and Determination of AST, ALT, ATP, ADP in Chlorogenic Acid, Forsythin, Baicalin and Wogonin on HL-7702Study revealed:compared with other groups, Forsythin combined with cefradine improve the content of AST、ALT, Wogonin combined with cefradine decreased the content of ATP、ADP.ConelusionShuanghuanglian freeze-dried powder for injection combined with cefradine for injection have certain toxic effect on HL-7702. the possible way are damaging cell membranes, increasing the express of HO-1、COX2and damaing mitochondrial. Forsythin damage cell membrane and wogonin damage mitochondrial are more significant than other group.