红景天苷对肝缺血再灌注损伤模型大鼠肝细胞自噬作用的影响

Salidroside in hepatic ischemia-reperfusion injury in rats and its induced autophagy reaction

目的观察红景天苷对肝缺血再灌注损伤（hepatic ischemia-reperfusion injury,HIRI）模型大鼠肝细胞自噬作用的影响。方法将90只大鼠按随机数字表法分为假手术组，模型组，红景天苷低、中、高剂量组，每组18只。低、中、高剂量组大鼠分别灌胃红景天苷水溶液7．5、15、30mg／kg，假手术组与模型组灌胃等体积生理盐水，1次／d，连续给药7d。除假手术组外，其余各组大鼠制备HIRI模型。分别于再灌注后4、8、16h取材，观察大鼠肝组织病理学改变，检测大鼠血清ALT、AST水平，采用Western Blot法检测缺血肝组织中自噬相关基因（autophagy-relatedgene，Beclin-1）蛋白及微管相关蛋白1轻链3-Ⅱ（microtubule-associate protein 1 lightchain3-Ⅱ，LC3-Ⅱ）表达；电镜观察各组肝细胞中自噬小体数量及形态。结果再灌注后4、8、16h，与模型组比较，红景天苷高剂量组大鼠血清Au[分别为（662．36±5．82）U／L比（983．67±8．96）U／L、（436．49±12．93）U／L比（1536．65±10．77）U／L、（168．61±8-34）U／L比（280．42±17.37）U／L]、AST[分另0为（513．29±11．74）U／L比（656．38±7．67）U／L、（276．29±9．21）U／L比（930．19±15．62）U／L、（97．83±4．29）U／L比（211．23±7．87）U／L]水平降低（P〈0．05）；再灌注后8、16h，红景天苷高剂量组LC3-Ⅱ[分别为（1．21±0．16）比（1．91±0．12）、（2．00±0．14）比（1．09±0．11）]、Beclin-Ⅱ分别为（3．53±0．19）比（7．15±0．14）、（2．65±0．27）比（7．60±0．21）]表达降低（P〈0．05）。再灌注后8h，红景天苷高剂量组电镜下肝细胞中自噬小体数量[（3．24±0．62）个比（7．84±O．45）个]降低（P〈0．05）。结论红景天苷对HIRI模型大鼠有保护作用，其作用机制之一可能与抑制肝细胞自噬有关。

Objective To research the effect and autophagy in hepatic ischemia-reperfusion injury based on relevant indicators of the specimens of rat liver which ischemia reperfusion model by salidroside pretreatment. Methods A total of 90 male SD rats were randomly divided into the sham group, the model group, the low, medium and high dose group, 18 rats in each group. The low, medium and high dose group rats were treated with 7.5, 15, 30 mg/kg salidroside solution by gavage, and the sham group and model group and model group were filled with saline in the same volume,one time per day. After 7 days, all the rats were set up with the model of IR except the rats in sham groups. The AST and ALT of serum, contrast between groups liver tissue by Optical microscope with HE dyeing at 4, 8, 16 h after reperfusion. Western Blot was used to detect the expression of protein of LC3 and Beclin-1. The number and morphology of autophagy in each group of liver cells were observed by electron microscopy. Results After reperfusion 4, 8, 16 h, the level of ALT （662.36 ± 5.82 U/L vs. 983,67 ± 8.96 U/L, 436.49 ± 12.93 U/L vs. 1 536 ± 10.77 U/L, 168.61 ± 8.34 U/L vs. 280.42 ±17.37 U/L） of the high dose group weresignificantly lower than the model group, and the AST （513.29±11.74 U/L vs. 656.38±7.67 U/L, 276.29± 9.21 U/L vs. 930.19 ± 15.62 U/L, 97.83 ± 4.29 U/L vs. 211.23 ± 7.87 U/L） of the high dose group were significantly lower than the model group. After reperfusion 8, 16 h, the expression of LC3-Ⅱ （1.21 ± 0.16 vs. 1.91 ± 0.12, 2.00 a： 0.14 vs. 1.09± 0.1l） in the high dose group were significantly lower than the model group, and the results were same to Beclinl （3.53 ± 0.19 vs. 7.15 ± 0.14, 2.65 ± 0.27 vs. 7.60 ± 0.21） （P〈0.05）. After reperfusion 8 h, the number of autophagosome （3.24± 0.62 vs.7.84 ± 0.45） in the high dose group were significantly lower than the model group （P〈0.05）. Conclusions The hepatic ischemia-reperfusion injury was serious, and inhibiting autophagy was one of possible mechanisms to protect liver cells by salidroside.