纳米姜黄素对高原环境急性肝损伤的保护作用及氧化应激的影响

Protective effect of Nanocurcumin on acute liver injury and oxidative stress of rats in high altitude environment

目的观察姜黄素及纳米姜黄素对高原大鼠急性肝损伤的保护作用。方法选择雄性SD大鼠80只,按随机数字表法分为平原对照组、高原对照组、普通姜黄素组、纳米姜黄素组,每组20只。普通姜黄素组和纳米姜黄素组将普通姜黄素粉末及纳米姜黄素溶解于0.9%氯化钠注射液中,分别按200 mg/kg和25 mg/kg的药物剂量灌胃;平原对照组和高原对照组均给予0.9%氯化钠注射液灌胃,均每日1次,连续3 d。将除平原对照组外的大鼠置于高原环境模拟舱制备高原急性肝损伤大鼠模型,舱内条件设置为12~12 h的光暗循环,白天12 h温度15℃,夜间12 h温度5℃,湿度(45±5)%。在舱内继续每日灌胃1次。于高原缺氧暴露48 h和72 h将大鼠麻醉后,取腹腔静脉血和肝脏组织,光镜下观察肝组织病理学改变,并比较各组血清肝功能酶学指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST),以及肝组织氧化应激指标过氧化氢酶(CAT)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)水平的差异。结果与平原对照组比较,高原对照组各时间点肝损伤病理评分、ALT、AST及MDA均明显升高〔暴露48 h:肝损伤病理评分(分)为3.90±0.20比1.91±0.16,ALT(U/L)为74.59±3.65比59.29±4.99,AST(U/L)为212.84±10.80比110.29±8.68,MDA(nmol/mg)为60.71±7.58比41.62±7.78;暴露72 h:肝损伤病理评分(分)为3.76±0.20比1.88±0.20,ALT(U/L)为72.99±4.32比60.05±4.97,AST(U/L)为207.12±7.43比104.71±8.31,MDA(nmol/mg)为58.61±7.08比43.31±6.42,均P<0.05〕,CAT、SOD及GSH水平明显下降〔暴露48 h:CAT(U/mg)为41.11±2.59比52.86±3.21,SOD(U/mg)为225.67±3.83比252.85±7.49,GSH(U/mg)为34.94±1.44比38.77±1.56;暴露72 h:CAT(U/mg)为42.91±2.72比53.40±2.71,SOD(U/mg)为228.14±6.31比255.67±7.09,GSH(U/mg)为36.30±2.08比40.15±2.45,均P<0.05〕;与高原对照组比较,普通姜黄素组和纳米姜黄素组肝损伤病理评分、ALT、AST及MDA水平均明显下降〔暴露48 h:肝损伤病理评分(分)为3.54±0.16、3.49±0.20比3.90±0.20,ALT(U/L)为65.10±5.54、64.50±5.28比74.59±3.65,AST(U/L)为193.60±9.09、194.00±10.89比212.84±10.80,MDA(nmol/mg)为48.12±5.64、48.28±7.40比60.71±7.58;暴露72 h:肝损伤病理评分(分)为3.48±0.12、3.42±0.18比3.76±0.20,ALT(U/L)为64.78±4.53、63.97±4.52比72.99±4.32,AST(U/L)为198.25±7.84、196.91±7.23比207.12±7.43,MDA(nmol/mg)为49.76±5.10、49.85±4.53比58.61±7.08,均P<0.05〕,CAT、SOD及GSH水平均明显增高〔48 h:CAT(U/mg)为45.86±2.09、45.72±2.35比41.11±2.59,SOD(U/mg)为240.34±4.24、239.86±5.20比225.67±3.83,GSH(U/mg)为38.24±1.11、38.21±1.34比34.94±1.44;72 h:CAT(U/mg)为46.31±3.50、46.35±3.41比42.91±2.72,SOD(U/mg)为240.90±4.06、240.43±5.98比228.14±6.31,GSH(U/mg)为38.38±2.40、38.29±1.68比36.30±2.08,均P<0.05〕,但普通姜黄素组与纳米姜黄素组间各指标比较差异均无统计学意义(均P>0.05)。结论纳米姜黄素与姜黄素预处理可缓解高原大鼠急性肝损伤程度,其作用可能与抑制氧化应激有关;且纳米姜黄素在用量明显少于普通姜黄素的情况下可发挥同样的肝损伤保护作用。

Objective To observe the protective effects of Curcumin and Nanocurcumin on acute liver injury in rats at high altitude.Methods A total of 80 male Sprague-Dawley(SD)rats were randomly divided into four groups:plain control,high altitude control,ordinary Curcumin and Nanocurcumin groups,with 20 rats in each group.In ordinary Curcumin and Nanocurcumin groups,the powders of ordinary Curcumin and Nanocurcumin were dissolved in 0.9%sodium chloride injectial solution,and gavaged at 200 mg/kg and 25 mg/kg drug doses,respectively;plain control group and high altitude control group were given 0.9%sodium chloride injection by gavage,once a day for 3 consecutive days.Except the plain control group,the rest of rats were placed in a simulated cabin at high altitude to prepare the rat model of acute liver injury at high altitude.The cabin conditions were set as light dark cycle for 12-12 hours,the temperature of 12 hours in the day time was 15℃,the temperature of 12 hours at night was 5℃,and the humidity was(45±5)%.The gavage in the cabin was continued,once a day.After the rats exposure to the hypoxia condition at high altitude for 48 hours and 72 hours,they were taken out from the cabin and anesthetized,then the celiac vein blood and liver tissue were taken to observe the histopathological changes of liver tissue under light microscope,and the differences between the index levels of liver function enzymes,alanine transaminase(ALT),aspartate transaminase(AST),oxidative stress indexes of liver tissue,catalase(CAT),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione(GSH)among the groups were compared.Results Compared with the plain control group,the histopathological liver-injury score,the levels of ALT,AST and MDA of the high altitude control group at each time point showed an obvious upward trend[exposure 48 hours:the histopathological liver-injury score was 3.90±0.20 vs.1.91±0.16,ALT(U/L)was 74.59±3.65 vs.59.29±4.99,AST(U/L)was 212.84±10.80 vs.110.29±8.68,MDA(nmol/mg)was 60.71±7.58 vs.41.62±7.78;exposure 72 hours:the histopathological liver-injury score was 3.76±0.20 vs.1.88±0.20,ALT(U/L)was 72.99±4.32 vs.60.05±4.97,AST(U/L)was 207.12±7.43 vs.104.71±8.31,MDA(nmol/mg)was 58.61±7.08 vs.43.31±6.42,all P<0.05],the CAT,SOD and GSH levels of the high altitude control group at each time point showed an downward trend[exposure 48 hours:CAT(U/mg):41.11±2.59 vs.52.86±3.21,SOD(U/mg):225.67±3.83 vs.252.85±7.49,GSH(U/mg):34.94±1.44 vs.38.77±1.56;72 hours:CAT(U/mg):42.91±2.72 vs.53.40±2.71,SOD(U/mg):228.14±6.31 vs.255.67±7.09,GSH(U/mg):36.30±2.08 vs.40.15±2.45,all P<0.05];compared with the high altitude control group,the histopathological liver-injury score,ALT AST and MDA levels in ordinary Curcumin group and Nanocurcumin group were decreased significantly[exposure 48 hours:the histopathological liver-injury score was 3.54±0.16,3.49±0.20 vs.3.90±0.20,ALT(U/L)was 65.10±5.54,64.50±5.28 vs.74.59±3.65,AST(U/L)was 193.60±9.09,194.00±10.89 vs.212.84±10.80,MDA(nmol/mg)was 48.12±5.64,48.28±7.40 vs.60.71±7.58;exposure 72 hours:the histopathological liver-injury score was 3.48±0.12,3.42±0.18 vs.3.76±0.20,ALT(U/L)was 64.78±4.53,63.97±4.52 vs.72.99±4.32,AST(U/L):198.25±7.84,196.91±7.23 vs.207.12±7.43,MDA(nmol/mg):49.76±5.10,49.85±4.53 vs.58.61±7.08,all P<0.05],CAT,SOD and GSH levels in ordinary Curcumin group and Nanocurcumin group were significantly increased[exposure 48 hours:CAT(U/mg)was 45.86±2.09,45.72±2.35 vs.41.11±2.59,SOD(U/mg)was 240.34±4.24,239.86±5.20 vs.225.67±3.83,GSH(U/mg)was 38.24±1.11,38.21±1.34 vs.34.94±1.44;exposure 72 hours:CAT(U/mg)was 46.31±3.50,46.35±3.41 vs.42.91±2.72,SOD(U/mg):240.90±4.06,240.43±5.98 vs.228.14±6.31,GSH(U/mg)was 38.38±2.40,38.29±1.68 vs.36.30±2.08,all P<0.05].However,there were no significant differences between ordinary Curcumin group and Nanocurcumin in terms of various indicators(all P>0.05).Conclusion Pretreatment with Curcumin and Nanocurcumin can alleviate the acute liver injury in rats at high altitude,which may be related to the inhibition of oxidative stress;in addition,Nanocurcumin has the same protective effect on liver injury when its dosage is significantly less than that of ordinary curcumin.