| Objective: Using rat liver cold ischemia from non cyclic reperfusion model ofimproved simulated liver in liver transplantation of preserved，In the animalexperiments in vitro of Compound Glycyrrhizin (CG) protective effect on livercold ischemia injury and of the high mobility group protein B1(HMGB1)regulation, and to investigate its possible mechanism, in order to furtherimprove the preservation of donor liver in liver transplantation and modifiedtransplant organs preservation solution provides the theory support.Method:100healthy adult SD rats were randomly divided into fivegroups. According preservation solution used for cold preservation wererandomly divided into CG-intervention group, sham operation group and thecontrol group， CG intervention group were HTK+CGI (1/100) group,HTK+CGI (1/75) group, HTK+CGI (1/50) group. The model of rat liver coldischemia modified using AIPRL model. Given the liver cold ischemia after12h0.4%trypan blue solution Determination of portal vein perfusion of hepaticmicrocirculation; Determination of enzyme activity and liver cold preservation12h perfused rat liver outflow liquid. Expression of inflammatory factors in ratswith cold preservation of liver12h after using enzyme linked immunosorbentassay; Determination of cell apoptosis in the liver tissue levels of rats with terminal transferase labeling method; Immunohistochemical techniquealtimetry mobility protein B1(HMGB1), Toll like receptor4(TLR4), nuclearfactor kappa binding (NF-κ b). Use SPSS Statistics19statistical software forstatistical analysis, statistical data in (x±s)expression, when P <0.01said theresults there are statistical differences.Results:1. The use of0.4%trypan blue solution given portal vein perfusionafter12h cold storage，to portal vein infusion, time of CGL group, CGM group,CGH group, hepatic left lateral lobe to uniform Aizen required were (83.9±2.3)S,(77.6±0.6) S,(73.5±1.1) S, obviously lower than the CON group (104±12.7) S (P<0.01), higher than that in SHAM group (41.6±2.8) S (P<0.01). Theintervention group, the CGH group was lower than that of CGM group, CGLgroup (P<0.01), CGM group than that in CGL group (P<0.01).2. Pathology: pathology scores in group CGL, group CGM, group CGH scoreswere (2.00±0.41),(1.20±0.46),(0.80±0.25) was significantly lower thanthat in CON group (2.60±0.51)(P <0.01), higher than that of SHAM group(0.20±0.12pathological score)(P <0.01). The intervention group had nostatistical difference.3. Results enzyme index shows: cold preservation after12h CGL group, CGMgroup, CGH group, ALT content in liver perfusion outflow liquid were:(155.10±17.36) U/L,(117.92±11.51) U/L,(47.96±20.99) U/L, significantly lowerthan that in CON group (244.76±71.59) U/L (P <0.01), higher than that inSHAM group (3.94±1.43) U/L (P <0.01); intervention group, CGH group than that in CGM group, CGL group (. P<0.01), the CGM group was lowerthan that of CGL group (the P<0.01). CGL group, CGM group, CGH group,AST content in liver perfusion outflow liquid were:(180.61±23.67) U/L,(135.65±20.74) U/L,(88.52±16.67) U/L, significantly lower than that inCON group (311.05±65.88) U/L (P <0.01), higher than that in SHAM group(9.46±1.06) U/L (P <0.05); intervention group, CGH group than that in CGMgroup, CGL group (P<0.01), the CGM group was lower than that of CGL group(P<0.01). In CGL group, CGM group, CGH group, LDH were (1513.70±154.23) U/L,(1190.60±127.87) U/L,(527.80±239.78) U/L, significantlylower than that in CON group (2549.70±631.76) U/L (P <0.01), higher thanthat in SHAM group (63.30±7.50) U/L (P <0.01); intervention group, CGHgroup than that in CGM group, CGL group (P<0.01), the CGM group waslower than that of CGL group (P<0.01).4. ELISA results show: Rat liver by in vitro cold ischemia preservation of12h,TNF-α in CGL group, CGM group, CGH in liver tissue of the group were133.09±45.86) pg/ml,(123.81.50±46.56) pg/ml,(115.63±44.28) pg/ml wassignificantly lower than that in CON group (243.89±25.16) pg/ml (P<0.01),but higher than that of SHAM group (53.56±4.74) pg/ml (P<0.05);intervention group, CGH group than that in CGL group (P<0.01). The contentof IL-6CGL group, CGM group, CGH in liver tissue of the group were:(110.51±13.78) pg/ml,(100.36±15.12) pg/ml,(73±16.64) pg/ml wassignificantly lower than that of CON group (157.71±13.28) pg/ml (P<0.01), but higher than that in SHAM group (35.13±9.76) pg/ml (P<0.01);intervention group, CGH group than that in CGL group (P<0.01).5. TUNEL results show： CGL group, CGM group, CGH group apoptosis celldensity were:(36.7±2.1) cell number/mm2,(30.3±3.4) cell number/mm2,(23.1±2.1) CON groups of cell number/mm2less than (42.6±2.1) cellnumber/mm2(P <0.01), but significantly lower than that in SHAM group (7.6±1.1) cell number/mm2(p <0.01). The intervention group, compared withthe CGH group (.P<0.01); CGL group compared with CGM group group(P<0.01).6. Immunohistochemistry showe,：each liver cold ischemia after12h liver cellsare NF-κ B p65protein expressed in different degrees, CGL group, CGMgroup, CGH group and NF-κ B expression p65protein index were:(6.33±1.45),(5.54±1.63),(3.17±1.52) lower than that of CON group (10.65±2.57P <0.01), but significantly higher than that in SHAM group and0.67±0.48(P<0.01). The intervention group, CGH group and CGL group (P <0.01).7. Each liver cold ischemia after12h TLR4expression in liver cells aredifferent, CGL group, CGM group, CGH group, TLR4protein expression indexis respectively:(5.26±1.10),(4.14±1.23),(2.41±0.55) higher than CONgroup (9.09±1.73)(P <0.01), but higher than that in SHAM group (0.23±0.45) and high (P <0.01). The intervention group, CGH group and CGL group(P <0.01).8. Each liver cold ischemia after12h HMGB1expression in liver cells are different, CGL group, CGM group, CGH group, the protein expression of TLR4index were:5.16±0.93,4.44±0.63,2.01±0.55lower than that of CON group8.09±1.73(P <0.01). The intervention group, CGL group, CGM group andCGH (P <0.01).Conclusions:1. Compound Glycyrrhizin intervention to ease the inflammatory injury ofliver cold ischemia time and improve liver cell function.2. Protective effect of Compound Glycyrrhizin may be through inhibition ofHMGB1–TLR4–NF-κB p65–TNF-a, IL-6expression to reduceinflammation injury of liver cold ischemia time... |
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