CAJ | 학술논문

目的观察小檗碱预处理减轻大鼠肝脏冷缺血再灌注(IR)损伤的作用,探讨磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/雷帕霉素靶蛋白(mTOR)信号通路在该作用中的分子机制.方法采用随即数字表法将雄性SD大鼠分为4组,小檗碱组(给予大鼠小檗碱100 mg·kg-·d-1灌胃2周后建立大鼠肝脏冷IR损伤模型)、二甲基亚砜(DMSO)组(操作同小檗碱组,用DMSO代替小檗碱)、IR组(操作同小檗碱组,用生理盐水代替小檗碱)及假手术组(给予生理盐水灌胃2周后行开腹和关腹处理).再灌注后3、6、24 h,通过血清学和组织学指标的检测,观察肝脏的损伤情况;比较各组肝细胞凋亡情况,检测PI3K/Akt/mTOR通路相关蛋白的表达情况.结果与假手术组比较,IR组和DMSO组肝细胞出现肿胀、坏死,肝脏氧化应激水平升高,细胞凋亡明显增加,Akt和mTOR磷酸化显著上调;小檗碱组大鼠肝功能得到明显改善,肝细胞肿胀、水肿减轻,缓解氧化应激损伤,肝细胞凋亡率显著降低,Bcl-2/Bax比值及caspase-3的表达水平显著升高,与此同时,Akt磷酸化也显著上调,而mTOR磷酸化水平相对降低.结论小檗碱可通过抑制肝脏氧化应激反应等途径缓解细胞大鼠肝脏后IR后的凋亡,改善大鼠肝功能,其机制与PI3K/Akt/mTOR信号通路的激活有关.
목적 관찰소벽감예처리감경대서간장랭결혈재관주(IR)손상적작용,탐토린지선기순3격매(PI3K)/단백격매B(Akt)/뢰파매소파단백(mTOR)신호통로재해작용중적분자궤제.방법 채용수즉수자표법장웅성SD대서분위4조,소벽감조(급여대서소벽감100 mg·kg-·d-1관위2주후건립대서간장랭IR손상모형)、이갑기아풍(DMSO)조(조작동소벽감조,용DMSO대체소벽감)、IR조(조작동소벽감조,용생리염수대체소벽감)급가수술조(급여생리염수관위2주후행개복화관복처리).재관주후3、6、24 h,통과혈청학화조직학지표적검측,관찰간장적손상정황;비교각조간세포조망정황,검측PI3K/Akt/mTOR통로상관단백적표체정황.결과 여가수술조비교,IR조화DMSO조간세포출현종창、배사,간장양화응격수평승고,세포조망명현증가,Akt화mTOR린산화현저상조;소벽감조대서간공능득도명현개선,간세포종창、수종감경,완해양화응격손상,간세포조망솔현저강저,Bcl-2/Bax비치급caspase-3적표체수평현저승고,여차동시,Akt린산화야현저상조,이mTOR린산화수평상대강저.결론 소벽감가통과억제간장양화응격반응등도경완해세포대서간장후IR후적조망,개선대서간공능,기궤제여PI3K/Akt/mTOR신호통로적격활유관.
Objective To confirm the protective effect of berberine (BBR) on cold ischemia reperfusion (I/R)-induced liver injury and to show whether the hepatic protection conferred by BBR involves the activation of phosphatidylinositol 3 kinase (PI3K) / protein kinase B (Akt)/mammalian target of rapamycin(mTOR) signal pathway.Method Adult male Sprague-Dawley rats were assigned randomly to four groups:BBR group (BBR was intragastrically administered at a dose of 100 mg·kg-1 · d-1 2 weeks before hepatic cold I/R treatment),dimethyl sulfoxide (DMSO) group (BBR was replaced by DMSO,and others were the same as BBR group),I/R group (BBR was replaced by normal saline,and others were the same as BBR group) and sham group (normal saline was administered 2 weeks before opening and closing abdomen treatment).Then the rats were sacrificed at 3,6,and 24 h after reperfusion.The liver function,oxidative stress level,apoptosis rate,and the expression of PI3K/Akt/mTOR related pathway proteins were assayed.Result As compared with sham group,the I/R-induced liver tissue displayed severe lobular distortion with widespread necrosis,high level of oxidative stress and apoptosis rate.As compared with I/R group,BBR dramatically attenuated the histopathologic damage,restored the liver function and decreased the oxidative stress level.Simultaneously,BBR significantly ameliorated the apoptosis by decreasing the apoptosis rate,increasing the Bcl-2/Bax ratio and inhibiting caspase-3 activity in rats subjected to hepatic I/R.The expression of p-Akt was effectively upregulated with the inhibited expression of p-mTOR.Conclusion Our result provides robust in vivo evidence that BBR can prevent I/R-induced oxidative stress and apoptosis.The mechanisms involved can be attributed to the activation of P]3K/Akt/mTOR signal pathway.