CAJ | 학술논문

目的探讨熊脱氧胆酸( Ursodeoxycholic acid,UDCA)通过减轻内质网应激保护链脲佐菌素诱导的糖尿病大鼠胰岛β细胞凋亡的作用.方法链脲佐菌素(50 mg/kg)一次性腹腔注射建立糖尿病大鼠模型(n=40),并将14只造模成功大鼠按随机数字表法随机分为糖尿病组7只,UDCA组7只,另取正常对照组10只.自造模成功第10天开始以UDCA(40 mg·kg-1·d-1)给大鼠灌胃30 d.饲养期间观察大鼠血搪.处死后留取大鼠血清和组织标本,测定空腹胰岛素,TUNEL检测胰岛β细胞凋亡的形态学改变.胰腺组织总RNA采用定制基因芯片对89条凋亡相关基因的表达进行检测,以RT-PCR和Western印迹法验证相关基因的表达.结果糖尿病大鼠血糖在给予UDCA治疗后逐渐下降,但仍然未降到正常水平.糖尿病大鼠空腹胰岛素降低,给予UDCA治疗后空腹胰岛素水平有所增加.TUNEL显示糖尿病组大鼠予UDCA治疗后减少了由链脲佐菌素引起的胰岛β细胞凋亡(P＜0.01).在89条基因中,糖尿病组上调基因42条,下调基因46条.UDCA可以逆转部分基因的影响.与对照组相比,糖尿病组大鼠Bax、Caspase-3、Bip、CHOPmRNA和CHOP蛋白显著上调(P＜0.05),而抗凋亡蛋白Bcl-2 mRNA显著下调(P＜0.05),给予UDCA治疗后这些参数均有明显改善(P＜0.05).结论熊脱氧胆酸可能通过减轻内质网应激达到保护链脲佐菌素诱导的糖尿病大鼠胰岛β细胞凋亡的作用.
목적 탐토웅탈양담산( Ursodeoxycholic acid,UDCA)통과감경내질망응격보호련뇨좌균소유도적당뇨병대서이도β세포조망적작용.방법 련뇨좌균소(50 mg/kg)일차성복강주사건립당뇨병대서모형(n=40),병장14지조모성공대서안수궤수자표법수궤분위당뇨병조7지,UDCA조7지,령취정상대조조10지.자조모성공제10천개시이UDCA(40 mg·kg-1·d-1)급대서관위30 d.사양기간관찰대서혈당.처사후류취대서혈청화조직표본,측정공복이도소,TUNEL검측이도β세포조망적형태학개변.이선조직총RNA채용정제기인심편대89조조망상관기인적표체진행검측,이RT-PCR화Western인적법험증상관기인적표체.결과 당뇨병대서혈당재급여UDCA치료후축점하강,단잉연미강도정상수평.당뇨병대서공복이도소강저,급여UDCA치료후공복이도소수평유소증가.TUNEL현시당뇨병조대서여UDCA치료후감소료유련뇨좌균소인기적이도β세포조망(P＜0.01).재89조기인중,당뇨병조상조기인42조,하조기인46조.UDCA가이역전부분기인적영향.여대조조상비,당뇨병조대서Bax、Caspase-3、Bip、CHOPmRNA화CHOP단백현저상조(P＜0.05),이항조망단백Bcl-2 mRNA현저하조(P＜0.05),급여UDCA치료후저사삼수균유명현개선(P＜0.05).결론 웅탈양담산가능통과감경내질망응격체도보호련뇨좌균소유도적당뇨병대서이도β세포조망적작용.
Objective To clarify the protective effect of nrsodeoxycholic acid ( UDCA ) on endoplasmic reticulum stress-mediated apoptosis in pancreatic β-cell of streptozotocin ( STZ )-induced diabetic rats.Methods Rats( n =40) received a single injection STZ( 50 mg/kg) intra-peritoneally and formed a β-cell injury model.Weight-matched normal rats( the control group,n =10 ) were injected with the buffer alone.STZ-treated rats with persistent random blood glucose higher than 16.7 mmol/L for 1 week were considered as diabetic status( n=14 ),then divided randomly into STZ-induced diabetes mellitus ( DM ) group ( n =7 ) and UDCA-treated DM group ( n =7 ).UDCA (40 mg· kg- 1,d-1 ) was administered daily by intragastric intubations throughout the experimental period (30 d).During the entire experiment,blood glucose in all rats was assessed.By the end of the experiment,all rats were sacrificed with the pancreas removed and the blood sample collected immediately.Fasting insulin levels were assayed by radioimmunoassay.The morphological changes of pancreatic β-cells apoptosis were determined by TUNEL assay.RNA in pancreas was abstracted and microarray containing 89 pieces of apoptosis related genes was applied.The related gene expressions were detected by RT-PCR and Western blot.Results The concentration of blood glucose in diabetic rats was gradually decreased after UDCA treatment,but at the end of the experiment it was still higher than that in the normal control group.The treatment with UDCA raised the fasting insulin level in diabetic rats,but this concentration was significantly lower as compared to the control group.Based on TUNEL stained tissue sections,the percentage of β-cell apoptosis of UDCA-treated DM group was significantly lower than that of STZ-induced diabetic group(P＜0.05 ).Among 89 genes,42 genes up-regulated and 46 genes down-regulated in diabetic rats,some of which were ameliorated by UDCA treatment.The expressions of Caspase-3,Bax,Bip,and CHOP mRNA in pancreas of DM group were significantly up-regulated as compared with those in the control group ( P ＜ 0.05 ) ; while the expression of Bcl-2 mRNA was markedly down-regulated (P＜0.05 ).However,these parameters in the U DCA-treated animals showed a marked improvement.Conclusion Ursodeoxycholic acid seems to protect pancreatic β-cell from apoptosis in STZ-induced diabetes by attenuating the severity of endoplasmic reticulum stress.