The Mechanism Of Lipid Related Genes On Different Medicines Preventing Gallbladder Cholesterol Gallstone

Aims:GD is one of the most common biliary diseases in abdominal surgery and the genes involved with lipid metabolism play the important roles in the development of GD.We attempted to detect the expression levels of lipid-metabolic genes during the different periods of LD feeding and analyze the discrepancies at the selected time points in order to reveal the possibility of a potential pathway existed.Furthermore,UDCA,YCHD,CPH,DCHD and EMO were utilized as therapeutic agents to feed the mice models of gallbladder cholesterol gallstone and we aimed to elucidate how the agents affect the expressions of lipid genes and the development of GD.Methods: After an adoptive feeding for 2W,C57BL/6J mice were randomized into 9 groups as below: i)0W,ii)4W,iii)8W,iv)RD,v)UDCA,vi)YCHD,vii)CPH,viii)DCHD,ix)EMO.Except for RD group that was fed with RD,the remaining groups were administrated with LD;the feeding time of previous two groups was 0W,4W,and other groups were 8W.Meanwhile,all mice groups were intervened with intragastricly equivoluminal distilled water,UDCA,YCHD,CPH,DCHD,EMO,respectively.Of note,the RD and 0W groups were served as “normal” controls,8W as model or GD control.Once the experimental periods for each of the groups ended,the blood,bile,gallstone were collected and liver tissues,gallbladder were harvested for further investigation.The concentrations of lipid profiles,chemical components of gallstone and CSI were examined.A portion of liver tissues was stained with HE for histological observation.Moreover,RT-PCR was performed to measure the relative expression levels of hepatic lipid genes,including Cyp7a1,Lxr,Abcb4,Cyp27a1,Abcg5,Cyp7b1,Abcb11,Srebp2,Abcg8,Hmgcr,Ppar m RNA.Results: In pre-test,the 100% of gallstone incidence was successfully obtained from the models mice.The gallstone incidence of 0W,RD,4W,8W group was 0,0,50% and 100%,respectively,while those of UDCA,YCHD,CPH,DCHD and EMO group were 30%,40%,40%,60% and 70%.The quality of gallbladder stone was identified as cholesterol gallstone.As feeding time progressed,the levels of BC,PL,TBA and CSI were shown an increasing tendency,additionally,there was a special peak concentration of TBA in 4W group,significantly higher than that in 0W and 8W groups.Conversely,in the UDCA,YCHD,CPH feeding groups,the levels of TC,PL,CSI decreased and TBA concentration increased.Thereafter,the histological observation of liver tissues demonstrated that there were no obvious abnormalities in 0W and RD groups.However,serious liver injury,such as cells arranging disorderly,fat droplets full of cytoplasm,suspicious changes of nucleo-cytoplasmic ratio were emerged at 8W group,the liver conditions in UDCA,YCHD,CPH,DCHD,EMO and 4W groups were mild and slight injury,which was better than that of 8W and less than RD group.RT-PCR detection indicated that Abcb4,Abcg5,Abcg8,Ppar m RNA were significantly increased and Srebp2,Cyp7b1,Cyp7a1 m RNA were decreased with lithogenic time going on.Strikingly,the transcriptional levels of Abcb11,Abcg8 and Cyp27a1 m RNA in UDCA group were notably increased,while the transcriptional levels of Ppar and Lxr were less than those in 8W group;the relative expressions of Cyp7a1,Cyp7b1,Lxr and Hmgcr m RNA in YCHD treatment group were approximately 3 folds higher than those in 8W group;CPH treatment may significantly decrease the expressions of Abcb4,Ppar and Lxr m RNA;the transcriptions of Cyp7a1 and Cyp27a1 m RNA in DCHD group were shown higher levels than those in 8W,while Srebp2 m RNA was decreased;under the administration of EMO treatment,Fxr,Cyp7a1,Cyp27a1,Ppar m RNA were increased to some extent.Conclusions: Changed expressions of Abcg5,Srebp2,Abcg8,Abcb4,Cyp7a1,Cyp7b1,Ppar m RNA have been related to the development of cholesterol gallstone.The experimental therapies of UDCA,YCHD and CPH have shown the effects on decreasing gallstone incidence,protecting liver function,reducing bile lipid composition.In addition,UDCA may regulate the expressions of Abcb11,Abcg8,Cyp27a1,Ppar,Lxr m RNA,eventually reversing the process of gallstones formation.Cyp7a1,Cyp7b1,Lxr and Hmgcr m RNA might be the effective targets for YCHD therepy.CPH might play a choleretic effect on down-regulating the expressions of Lxr,Abcb4,Ppar m RNA.DCHD showed significant influence on the transcriptions of Cyp7a1 and Cyp27a1,Srebp2 m RNA.The administration of EMO might inhibit the formation of gallstone via regulating Fxr,Cyp7a1,Cyp27a1 and Ppar m RNA.