基于分子对接和内质网应激探讨老鼠簕生物碱A对小鼠急性肝损伤的作用

The effect of Acanthus ilicifolius alkaloid A on acute liver injury mice based on endoplasmic reticulum stress and molecular docking

目的:探讨老鼠簕生物碱A(HBOA)对脂多糖(LPS)所致小鼠急性肝损伤的作用及机制。方法:采用AutoDock Tools软件将HBOA与GRP78、IL-1β、TNF-α、IL-6蛋白进行分子对接。腹腔注射LPS(10 mg/kg)建立小鼠急性肝损伤模型,分为正常组、模型组、联苯双酯阳性药组及HBOA高、中、低剂量组。连续灌胃给药10 d,除正常组外,其余5组小鼠均腹腔注射LPS,诱导急性肝损伤模型。检测各组肝组织中乳酸脱氢酶(LDH)、谷胱甘肽(GSH)水平,免疫组化法检测白细胞介素(IL)-1β的表达,Western blotting法测定内质网应激相关蛋白葡萄糖调节蛋白78(GRP78)、C/EBP同源体蛋白(CHOP)、Caspase-12及炎症相关蛋白肿瘤坏死因子(TNF)-α、IL-6的表达。结果:HBOA与GRP78、IL-1β、TNF-α、IL-6蛋白之间的最优对接构象结合能分别为-5.77 kcal/mol、-5.48 kcal/mol、-5.83 kcal/mol、-5.36 kcal/mol。与正常组相比,模型组小鼠肝组织中LDH含量及GRP78、CHOP、Caspase-12、IL-1β、TNF-α和IL-6表达显著增高,GSH水平显著降低(均P<0.05)。与模型组相比,HBOA高、中剂量组和阳性药组LDH含量及GRP78、CHOP、Caspase-12、IL-1β、TNF-α和IL-6表达明显降低,GSH水平显著升高(均P<0.05)。结论:HBOA能在一定程度上减轻LPS所致的小鼠急性肝损伤,其保肝机制可能与减轻炎症、减少内质网应激、抑制肝细胞凋亡相关。

Objective:To investigate the effects and mechanisms of Acanthus ilicifoliusalkaloid A(HBOA)on endoplasmic reticulum stress(ERS)inlipopolysaccharide(LPS)-induced acute liver injury in mice.Methods:Molecular docking between HBOA with GRP78 or IL-1βor TNF-αor IL-6 protein was performed using AutoDock Tools.Acute liver injury mouse model was established by intraperitoneal injection of LPS(10 mg/kg).Six groupswere set up as follows:the normal group,model group,positive control group(Bifendate),and HBOA high-,medium-,and low-dose groups.Mice were given intragastric administration for 10 days.Except for the normal control group,the rats in other five groups were given LPS via intraperitoneal injection to induce acute liver injury.The LDH and GSH levels in liver tissue were determined.The immunohistochemical method was performed to detect the expression of IL-1β.Western blotting analysis was used to determine the expressionlevels of GRP78,CHOP,Caspase-12,TNF-αand IL-6 proteins.Results:The molecular docking result showed that the binding energy of optimal conformational between HBOA and GRP78 or IL-1βor TNF-αor IL-6 were-5.77 kcal/mol,-5.48 kcal/mol,-5.83 kcal/mol and-5.36 kcal/mol,respectively.Compared with the normalgroup,the LDH content and the expression levelsof GRP78,CHOP,Caspase-12,IL-1β,TNF-αand IL-6 protein were increased significantly,while the GSH level was decreased in the model group(P<0.05).Compared with the model group,the LDH content and the expression of GRP78,CHOP,Caspase-12,IL-1β,TNF-αand IL-6 protein were decreased,whereas the GSH level was notably increased in the positive control group,high-dose and middle-dose HBOA groups(P<0.05).Conclusion:HBOA can reduce the degree of liver injury caused by LPS in mice to some extent,the mechanism of its protective effect on liver may be related to alleviate the inflammation,and the ability of reducing ERS and liver cell apoptosis.