铁皮石斛无菌纳米粉抗肝损伤作用及机制

徐博; 吴畏难; 赵丽晶; 陈雪

doi:10.11847/zgggws1124478

铁皮石斛无菌纳米粉抗肝损伤作用及机制

Effect and mechanism of sterile Dendrobium officinale nanoparticles on liver injury in mice

摘要

摘要:
目的探讨铁皮石斛无菌纳米粉抗肝损伤的药理作用并探讨其可能的机制。
方法 50只小鼠随机分为对照组、模型组、铁皮石斛纳米粉低、高剂量组和铁皮石斛普通粉组。试剂盒法检测小鼠血清碱性磷酸酶(ALP)、乙醇脱氢酶（ADH）活性和总抗氧化能力(T-AOC)，肝组织谷胱甘肽过氧化物酶（GPx）、还原型谷胱甘肽(GSH)、过氧化氢酶(CAT)活力；利用流式细胞仪检测肝细胞凋亡率；免疫组化法检测肿瘤坏死因子-α（TNF-α）表达；蛋白质免疫印迹法(WB)检测肝细胞Bcl-2、Bax、caspase-3、caspase-8蛋白表达。
结果与模型组比较，高剂量铁皮石斛纳米粉组小鼠血清ALP、ADH活力分别为（31.82 ± 3.35）U/L、（54.37 ± 2.89）U/mL均明显降低（P < 0.01），T-AOC活力（8.30 ± 2.06）U/mL明显升高（P < 0.05）；肝组织CAT、GPx活力和GSH含量分别为（68.56 ± 4.07）U/mg、（523.65 ± 32.67）U/mg、（26.28 ± 3.98）μmol/g均明显升高（P < 0.01）；肝细胞凋亡率明显下降（P < 0.01）,肝组织中TNF-α表达下调（P < 0.01），Bcl-2蛋白表达显著增加（P < 0.01），Bax、caspase-3、8蛋白表达显著减少（P < 0.01，P < 0.001）。与铁皮石斛普通粉组比较，铁皮石斛纳米粉组小鼠各项指标改善更为明显。
结论铁皮石斛无菌纳米粉对小鼠肝损伤的保护作用明显优于普通粉，其作用机制可能与抗氧化损伤、减轻肝细胞凋亡、抑制炎性细胞因子表达有关。

Abstract:
Objective To explore pharmacological effect and mechanism of sterile Dendrobium officinale nanoparticles (DON) on liver injury.
Methods Fifty Kunming mice were randomly divided into a normal control group, model group, low- and high-DON groups (LDON and HDON with gastric gavage of DON at doses of 0.125 and 0.250 g/kg once a day continuously for 7 days), and common Dendrobium officinale powder group (DOP with DOP of 0.250 g/kg). Levels of alkaline phosphatase (ALP), alcohol dehydrogenase (ADH) and total antioxidant capacity (T-AOC) in plasma, the glutathione peroxidase (GPX) activity, glutathione (GSH) and catalase (CAT) contents in liver tissues were detected with kit assay. Cell apoptosis rate in hepatocyte suspension was determined with flow cytometry. Expression of tumor necrosis factor-α (TNF-α) was detected with immunohistochemistry. The expression of B cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), caspase-3 and caspase-8 in liver tissues were detected with Western blot.
Results Compared with those in the model group, the levels of of serum ALP (31.82 ± 3.35 U/L) and ADH (54.37 ± 2.89 U/mL) was significantly decreased (both P < 0.01) and T-AOC capability (8.30 ± 2.06 U/mL) was improved (P < 0.05) in HDON group; the activity of CAT (68.56 ± 4.07 U/mg), GP_X (523.65 ± 32.67 U/mg) and GSH (26.28 ± 3.98 μmol/g) in liver tissues were increased (all P < 0.01); the apoptosis rate of hepatocytes was significantly decreased (P < 0.01); in liver tissues, the expression of TNF-α was significantly down-regulated (P < 0.01); Bax, caspase-3 and caspase-8 protein expression decreased (both P < 0.01) but bcl-2 (1.13 ± 0.12) protein expression increased (P < 0.01). Compared with those of the DOP group, the improvements in all the indexes of the DON group were more obvious.
Conclusion The protective effect of DON on liver injury in mice is significantly better than that of DOP, and the mechanism of the effects may be related to antioxidant damage, inhibited expressions of inflammatory cytokines and reduced liver cell apoptosis.

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