| Anti-inflammatory drugs are consumed in the world,second only to antibiotics.Commonly used anti-inflammatory drugs in clinic are steroids and non-steroidal anti-inflammatory drugs,but there are many adverse reactions.Traditional Chinese medicine anti-inflammatory drugs have a long history of application,abundant resources,and fewer adverse reactions,causing widespread concern.Drug Y is a hospital preparation that has good anti-inflammatory and antibacterial activity.The drug is not resistant to clinical pathogens and drug resistance,but its efficacy and mechanism of action have not been systematically studied.In this part of the study,Y drugs were used as research objects.The SD rat and KM mice,RAW264.7 cells and clinical pathogens were studied to investigate the in vitro and in vivo responses of Y drug to acute and chronic inflammation.The effects of anti-inflammatory mechanisms and antibacterial activity are as follows:1.Pharmacology Research:The safety and maximum gavage dose of Drug Y were tested with acute toxicity test.Meanwhile,the anti-acute and chronic inflammatory activity of Drug Y were tested with four classical models of mice or rat,namely xylene-induced ear swelling test,cotton pellet-induced granuloma test,acetic acid-induced vascular permeability test and carrageenan-induced paw edema test.The results demonstrated that the maximum dose of Drug Y is 100mg/kg and the weight of mice in the Drug Y group significantly increased.Also,Drug Y could increase the mouse thymus index,inhibition of atrophy of the spleen.Drug Y significantly reduced mouse ear swelling,peritoneal capillary permeability increased in rats,cotton ball granuloma and the best time in inhibiting toe swelling was 3 h.The toxicity of Drug Y is low,and Drug Y had a good therapeutic effect on acute and chronic inflammation.2.Animal anti-inflammatory mechanism:The influences of Drug Y on LPS-induced ALI mice were studied through testing the changes in the pathological of lung tissue in mice,the contents of IL-1β,PGE2 in serum,the protein expression of COX-2,NLRP3,p65 and the phosphorylate of p65 in lung tissue.The results demonstrated after pre-treatment Drug Y,inflammatory infiltration and erythrocyte proliferation in lung tissue were relieved,and alveolar structural integrity in lung tissue increased.At the same time,the levels of IL-1βand PGE2 in serum and the protein expressions of COX-2,NLRP3 and phosphorylated p65 expression in lung tissue were inhibited.Drug Y could regulate the expression of inflammatory proteins and the secretion of inflammatory cytokines by affecting the NF-κB signaling pathway,relieve lung tissue damage and exert anti-inflammatory effects in vivo.3.Cells anti-inflammatory mechanism:The activity of RAW264.7 cells were detected by MTT assay,and the dose of 08 mg/mL were safety dosage.At 12h after LPS induction,the model of RAW264.7 cells inflammation was the best.The influences of Drug Y on LPS-induced RAW264.7 cells inflammatory response in vitro were studied through the changes in the contents of IL-1β,PGE2 in cell supernatant,the protein expression of COX-2,NLRP3,p65 and the phosphorylate of p65 in RAW264.7 cells.The results showed that after treatment with Drug Y,the levels of IL-1βand PGE2 in cell supernatant decreased.The protein expressions of COX-2,NLRP3 and phosphorylated p65 expression in RAW264.7 cells were inhibited.It all shows that Drug Y had a low cytotoxicity.And the anti-inflammatory activity of Drug Y in vitro was related to NF-κB signaling pathway,which could relieve inflammation by inhibiting the secretion of inflammatory cytokines and inflammatory proteins.4.Antibacterial test:The anti-bacterial activity of Drug Y were studied through testing the diameter of inhibition zone,the minimum inhibitory concentration（MIC）and the bacterial growth curve after Drug Y treatment.The inhibition effect of Drug Y on Staphylococcus aureus is the most obvious,the diameter of inhibition zone is 26.04±0.87 mm.The inhibitory effect on Escherichia coli,Enterococcus faecalis,Pseudomonas aeruginosa,and Streptococcus pneumoniae was not obvious.After Drug Y treatment 0-4 h,the growth of Staphylococcus aureus was significantly inhibited,and MIC was 25 mg/mL.Acute liver failure（ALF）is a multi-factorial clinical syndrome of critically ill patients with liver dysfunction.Oxidative stress and inflammatory response mediate the occurrence and development of ALF,and play an important role in ALF.This study investigated the protective effect of TAENN in ALF mice.Details are as follows:1.The influences of TAENN on LPS/Gal N-induced ALF mice were studied through testing the changes in the survival curve of mice,the pathological of liver tissue in mice,liver index,the contents of aminotransferase,SOD,GSH,MDA,ROS,CAT,GSH-Px in serum.The results demonstrated after pre-treatment TAENN,miec mortality and the liver microstructure alleviated.The contents of AST,ALT,MDA in serum markablely decreased.The contents of GSH,SOD in serum increased significantly.Meanwhile,the contents of ROS,MDA in liver tissue decreased,and the levels of GSH,SOD,CAT,GSH-Px in liver tissue sharply increased.It shows that TAENN could inhibit oxidative stress in ALF mice.2.The influences of TAENN on LPS/Gal N-induced ALF mice were studied through testing the changes in the levels of TNF-α,IL-1β and IL-6 in serum,the contents of TNF-α,IL-1β,IL-6,PGE2 and NO in liver tissue,the m RNA expressions of TNF-α,IL-1β,IL-6,i NOS and COX-2 in liver tissue,the protein expressions of i NOS,COX-2,p65 and the phosphorylate of p65 in liver tissue.The results showed that after administration of TAENN,the contents of TNF-α,IL-1β and IL-6 in serum or liver tissue and the contents of NO and PGE2 in liver tissue were significantly decreased,the m RNA expressions of TNF-α,IL--6,i NOS,COX-2 reduced,the protein expressions of i NOS,COX-2 and phosphorylated p65 were inhibited.It proved that TAENN could inhibit inflammatory response. |
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