The Relationship Between BSEP/MRP2Changi-Ng And Cholestasis In Cirrhotic Rat Liver After Ischemia-Reperfusion

Objective: Through the cirrhotic rat bile producing rate bile (TB, CB) contents serum(AST, ALT, TB, CB) contents and Bsep/Mrp2proteins expression changeing after pringlemanenuer liver ischemia-reprfuion,to reveal part of molecular mechanism of cholestasis happenedafter liver pringle manenuer ischemia-reperfusion in cirrhosis rats.Methods: Experiment divided into three groups:group A:nomal rats liver pringle manenuerischemia-reperfusion;groupB:cirrhotic rats Sham operation group; Group C: cirrhotic rats pringlemanenuer ischemia-reperfusion group,each group collect relevant laboratory specimen(the Firstday、the third day、the five day) after operation respectively.In.personalized medicine way,takingSD rats TAA to make cirrhotic rats model,after12weeks, models production were completed.wecollected each experimental rat1hour of bile contents4ml Venous blood and liver tissuespecimens,calculated each rat bile producing rate,conventional biochemical analysis was used forbile (TB, CB)contents and serum (AST, ALT, TB, CB) contents. analysis Each group Bsep/Mrp2expression on different time points analysed by Western blotting and Immunohistochemistry.Results:1Cirrhotic rats animal model production:30rats were used for liver cirrhosis modelestablishment,after12weeks,models had been established by drinking TAA,number of successwere25,failures5,no deaths, Success rate88.3%.Cirrhotic liver appears texture partial hard coloruneven appear nodules and surface roughness.2Liver ischemia-reperfusion model established:Group A:12normal rats were used for models established situations,Successes11（91.7%),deaths1（8.3%).Group B (shamming operation group):12cirrhotic rats were used for models establishedsituations,Successes12(100%),deaths0(0%).Group C:12cirrhotic rats were used for models established situations,Successes9(75%),deaths3(15%). 3Bile producing rate：the first day after surgery,C compared with A (*P<0.01),C andB(*P>0.01), reached the third day after surgery A and B compared with C (P<0.01),but after thefifth days of operation,the comparison of A and B with C (*P>0.01).(P<0.01have Statisticaldifferences)4Biochemical results:(1) TB and CB contents of bile:the(first third)days after operation,A compare with C haveStatistical differences (▲P<0.01).The third after surgery C compare with B(﹡P<0.05),but Ccompare with B on the (first fifth)day after surgery and with A on the fifth day have no Statisticaldifferences.(▲P<0.01﹡P<0.05have Statistical differences)(2) Serum ALT, AST, TB, CB content：a、ALT:A and C (1day3day)((P<0.05),the five day after operation (▲P<0.01).B and Ccomparison (1d,3d,5d)were no statistically significant differences.b、AST:the first day after operation A and B compare with C (▲P<0.01),reach the third day A andC comparison (▲P<0.01),but B and C （▲P>0.01).The five day after Operation A and Bcompare with C (▲P>0.01).c、TB:A and B compare with c (1day3day5day)(▲P<0.01)d、CB:A and B compare with c (1day3day5day)(▲P<0.01)(*P<0.05)5Western bloting analysis results are shown:appear protein banding at160KDa and190KDa,meanwhile,at43KDa presents control banding.In the same conditions of control opticaldensity value;(1) Different groups the same time point Bsep expression:the (first third) day after surgery,theoptical density value of C group (30.33±2.18,25.40±3.97) compare with A(56.52±1.92,50.71±2.92)and B (40.85±4.35、35.33±2.51) reduced significantly,C groupcompared with A and B P<0.01, the fifth day after surgery C group (24.61±1.05) and A(50.17±4.71）comparison P <0.01,but C compare with B(30.23±2.12)P>0.01.(2) Different groups the same time point Mrp2expression:the (first third) day after surgery,theoptical density value of C group (28.71±2.79,26.12±3.15) compare with A (43.51±3.48，42.54±3.56) and B (33.52±2.09,29.71±3.06) reduced Significantly, C group compared with A andB P<0.01, among,.the fifth day after surgery C group (30±3.45) and A (50.17±4.71) P<0.01,Ccompare with B (35±2.43) P<0.01.6Immunohistochemical analysis results: different groups the same time points Bsepexpression:Tthe experiment result shows:Bsep expression decreased after liver cirrhosis and/orischemia-reperfusion,meanwhile,showing Bsep have tendency to shift into cells.The results of each group:the (first third fifth) day after operation, C compare with A P<0.01. The third day C and Bcomparison (p<0.01),B and A (P<0.01.). The fifth day C and B comparison p>0.01.Conclusions:Through cirrhotic rats liver bile producing rate bile(TB CB) contents andserum(ALT AST TB CB) contents after pringle manenuer ischemia and reperfusion compare withControl groups,indicating cirrhotic liver tolerance decrease significantly,after reperfusion liverfunction easy to disorder and cause cholestasis occurs,whicih often the serious more compare withnormal live constandly,meanwhile, after cirrhotic pringle manenuer liver ischemia-reperfusion,theassociation between Bsep and Mrp2expression decrease and the degree of cholestasis happeningare corresponded.