OBJECTIVE:To dynamically observe the efficacy of Jieduan Niwan formula(JDNW)on a rat model of acute-on-chronic liver failure(ACLF).METHODS:Seventy Wistar rats were divided into control group(6 rats),model group(22 rat...OBJECTIVE:To dynamically observe the efficacy of Jieduan Niwan formula(JDNW)on a rat model of acute-on-chronic liver failure(ACLF).METHODS:Seventy Wistar rats were divided into control group(6 rats),model group(22 rats),JDNW group(21 rats),and SP600125 group(21 rats).13 weeks'porcine serum injection followed with D-galactosamine and lipopolysaccharide joint acute attack was used to establish ACLF model.Rats in JDNW group were orally given JDNW formula for 3 days before acute attack;rats in SP600125 group were injected with SP60012530 min ahead of acute attack.Rats were sacrificed respectively at 4,8 and 12 h after model established.Alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),Creatinine(CR),blood urea nitrogen(BUN),prothrombin activity(PTA)were examined by biochemical process,Tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),transformed growth factor-beta 1(TGF-β1),High mobility group box-1(HMGB-1),CD3,CD4,CD8 were analyzed by enzyme-linked immunosorbent assay,apoptotic index(AI)was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining,expression of Bad,phosphorylated Jun N-terminal kinases(p-JNK)and Cytochrome C(Cyt C)were detected by immunohistochemical analysis,Bax and Bid were detected by Western blot analysis.RESULTS:In model group,the levels of ALT,AST,TBIL,CR,BUN,IL-1β,IL-6,IL-10,TGF-β1 and HMGB-1 remarkably increased and PTA decreased compared with control group(P<0.05),as time goes on,ALT,AST,TBIL,CR,BUN,continued to grow,while IL-1β,IL-6,IL-10,HMGB-1,TGF-β1 and PTA gradually decreased;massive necrosis could be seen;the levels of TNF-α,CD3,CD4,CD8,AI,p-JNK,Bax,Bad,Bid and Cyt C increased at 4 h and peaked at 8 h,but decreased at 12 h(P<0.05).JDNW group,by contrast,showed less pathological injury,increased PTA level,and reduced ALT,AST,TBIL,TNF-α,IL-1β,IL-6,IL-10,TGF-β1,HMGB-1,CD3,CD4 and CD8 levels(P<0.05),moreover,the AI and expression of p-JNK,Bax,Bad,Bid and Cyt C were lower than model group at 4 and 8 h but were higher at 12 h(P<0.05).Similar results were observed in SP600125 group.CONCLUSION:An ACLF rat model with low mortality can be established by porcine serum joint with D-galactosamine+lipopolysaccharide induction;JDNW decoction can effectively suppress the inflammatory reaction,improve the immune system,and protect the liver of ACLF rats,the mechanism might involve the inhibition of the JNK-induced mitochondrial apoptotic pathway.展开更多
OBJECTIVE: To investigate the protective effects and underlying mechanism of Qingdu decoction(QDD) on experimental rats with severe liver injury induced by thioacetamide(TAA).METHODS: A total of 40 Wistar rats were ra...OBJECTIVE: To investigate the protective effects and underlying mechanism of Qingdu decoction(QDD) on experimental rats with severe liver injury induced by thioacetamide(TAA).METHODS: A total of 40 Wistar rats were randomly divided into normal group(n = 10) and experimental group(n = 30). Rats were administrated the same content of saline in normal group. The rats inthe experimental group were pretreated with TAA at dose of 12 mg/kg lasting 8 weeks, and from 9th week to 12 th week, with TAA at concentration of 36mg/kg. During the 9th week to 12 th week period,the rats were randomly divided into three subgroups(n = 10 each) simultaneously based on the treatment categories: model group, lactulose(LA,3.5 m L/kg) group and QDD(5.95 g/kg) group, orally once per day respectively. At the 12 th week, the content of serum alanine transaminase(ALT), aspartate transaminase(AST), total bilirubin(TBIL), endotoxin(ET) and tumor necrosis factor α(TNF-α) was detected by automatic biochemical analyzer. The plasma prothrombin time(PT), prothrombin time-international normalized ratio(PTR) and prothrombin time activity(PTA) were measured by automatic coagulation analyzer. The level of lipopolysaccharide(LPS)-binding protein(LBP), cluster differentiation 14(CD14) and Toll-like receptor 4(TLR4) expressions was measured by both western blot(WB) and real-time polymerase chain reaction(real-time PCR).RESULTS: Compared with the model group, hepatic morphology in the QDD group was improved under light microscope and transmission electron microscope; at the same time, the contents of serum ALT, AST, TBIL, ET and TNF-α, and level of LBP, CD14 and TLR4 expressions in liver tissues were significantly decreased compared with the model group(P < 0.05), while PTA in the QDD group was enhanced(P < 0.05).CONCLUSION: QDD has the functional effect on improving the injured liver through inhibiting the LPS/TLR4 signaling pathway thus decreasing the level of the inflammatory medicators.展开更多
基金National Natural Science Foundation of China:Mechanisms of Heprescription of Truncation and Inverse Draft on Preventing and Treating Acute-on-Chronic Liver Failure Based on E2F1 Induced Hepatocytic Proliferation and Apoptosis(No.81573767)。
文摘OBJECTIVE:To dynamically observe the efficacy of Jieduan Niwan formula(JDNW)on a rat model of acute-on-chronic liver failure(ACLF).METHODS:Seventy Wistar rats were divided into control group(6 rats),model group(22 rats),JDNW group(21 rats),and SP600125 group(21 rats).13 weeks'porcine serum injection followed with D-galactosamine and lipopolysaccharide joint acute attack was used to establish ACLF model.Rats in JDNW group were orally given JDNW formula for 3 days before acute attack;rats in SP600125 group were injected with SP60012530 min ahead of acute attack.Rats were sacrificed respectively at 4,8 and 12 h after model established.Alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),Creatinine(CR),blood urea nitrogen(BUN),prothrombin activity(PTA)were examined by biochemical process,Tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),transformed growth factor-beta 1(TGF-β1),High mobility group box-1(HMGB-1),CD3,CD4,CD8 were analyzed by enzyme-linked immunosorbent assay,apoptotic index(AI)was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining,expression of Bad,phosphorylated Jun N-terminal kinases(p-JNK)and Cytochrome C(Cyt C)were detected by immunohistochemical analysis,Bax and Bid were detected by Western blot analysis.RESULTS:In model group,the levels of ALT,AST,TBIL,CR,BUN,IL-1β,IL-6,IL-10,TGF-β1 and HMGB-1 remarkably increased and PTA decreased compared with control group(P<0.05),as time goes on,ALT,AST,TBIL,CR,BUN,continued to grow,while IL-1β,IL-6,IL-10,HMGB-1,TGF-β1 and PTA gradually decreased;massive necrosis could be seen;the levels of TNF-α,CD3,CD4,CD8,AI,p-JNK,Bax,Bad,Bid and Cyt C increased at 4 h and peaked at 8 h,but decreased at 12 h(P<0.05).JDNW group,by contrast,showed less pathological injury,increased PTA level,and reduced ALT,AST,TBIL,TNF-α,IL-1β,IL-6,IL-10,TGF-β1,HMGB-1,CD3,CD4 and CD8 levels(P<0.05),moreover,the AI and expression of p-JNK,Bax,Bad,Bid and Cyt C were lower than model group at 4 and 8 h but were higher at 12 h(P<0.05).Similar results were observed in SP600125 group.CONCLUSION:An ACLF rat model with low mortality can be established by porcine serum joint with D-galactosamine+lipopolysaccharide induction;JDNW decoction can effectively suppress the inflammatory reaction,improve the immune system,and protect the liver of ACLF rats,the mechanism might involve the inhibition of the JNK-induced mitochondrial apoptotic pathway.
基金Supported by Beijing Natural Science Foundation of China(Investigation of the Mechanism on Qingdu Decoction in Repairing Injured Liver with TAA in Rat Based on Decreasing Intestinal Permeability,No.7142023)
文摘OBJECTIVE: To investigate the protective effects and underlying mechanism of Qingdu decoction(QDD) on experimental rats with severe liver injury induced by thioacetamide(TAA).METHODS: A total of 40 Wistar rats were randomly divided into normal group(n = 10) and experimental group(n = 30). Rats were administrated the same content of saline in normal group. The rats inthe experimental group were pretreated with TAA at dose of 12 mg/kg lasting 8 weeks, and from 9th week to 12 th week, with TAA at concentration of 36mg/kg. During the 9th week to 12 th week period,the rats were randomly divided into three subgroups(n = 10 each) simultaneously based on the treatment categories: model group, lactulose(LA,3.5 m L/kg) group and QDD(5.95 g/kg) group, orally once per day respectively. At the 12 th week, the content of serum alanine transaminase(ALT), aspartate transaminase(AST), total bilirubin(TBIL), endotoxin(ET) and tumor necrosis factor α(TNF-α) was detected by automatic biochemical analyzer. The plasma prothrombin time(PT), prothrombin time-international normalized ratio(PTR) and prothrombin time activity(PTA) were measured by automatic coagulation analyzer. The level of lipopolysaccharide(LPS)-binding protein(LBP), cluster differentiation 14(CD14) and Toll-like receptor 4(TLR4) expressions was measured by both western blot(WB) and real-time polymerase chain reaction(real-time PCR).RESULTS: Compared with the model group, hepatic morphology in the QDD group was improved under light microscope and transmission electron microscope; at the same time, the contents of serum ALT, AST, TBIL, ET and TNF-α, and level of LBP, CD14 and TLR4 expressions in liver tissues were significantly decreased compared with the model group(P < 0.05), while PTA in the QDD group was enhanced(P < 0.05).CONCLUSION: QDD has the functional effect on improving the injured liver through inhibiting the LPS/TLR4 signaling pathway thus decreasing the level of the inflammatory medicators.
