目的 基于沉默信息调节因子1(silent information regulator 1，SIRT1)/高迁移率族蛋白B1(high mobility group protein B1，HMGB1)/核转录因子-κB(nuclear transcription factor-KB，NF-κB)信号轴探讨丹参绞股蓝茶对四氯化碳(carbon tetrachloride，CCl4)所致小鼠急性肝损伤肝细胞炎症凋亡反应的抑制作用和机制研究。方法 C57BL/6小鼠随机分为正常组、模型组、白藜芦醇组、丹参绞股蓝茶低、中、高剂量组、阳性药组，连续灌胃给药14 d。采用腹腔注射0。5％CCl4橄榄油溶液(5 mL/kg)的方法建立小鼠急性肝损伤模型。生化法检测小鼠血清中丙氨酸转移酶(alanine transferase，ALT)、天冬氨酸转移酶(aspartate transferase，AST)、乳酸脱氢酶(lactate dehydrogenase，LDH)及肝组织羟脯氨酸(hydroxyproline，Hyp)、丙二醛(malonaldehyde，MDA)和超氧化物歧化酶(superoxide dismutase，SOD)水平;酶联免疫吸附法检测血清炎性因子肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素-6(interleukin-6，IL-6)和白细胞介素-1β(interleukin-1β，IL-1 β)水平;苏木素-伊红(HE)染色和TUNEL染色检测肝组织病理形态结构和肝细胞凋亡情况;Western Blot检测SIRT1、HMGB1和NF-κB蛋白表达。结果 与正常组比较，模型组小鼠血清ALT、AST、LDH水平和肝组织Hyp活性显著升高，以及肝组织中MDA和SOD活性明显降低，血清炎性因子TNF-α、IL-6和IL-1β水平显著升高(P＜0。05或P＜0。01)，肝组织存在明显的病理损伤以及肝细胞凋亡情况，肝组织中SIRT1蛋白的表达明显降低，HMGB1和NF-κB蛋白的表达升高;与模型组比较，丹参绞股蓝茶高剂量组和白藜芦醇组小鼠血清肝功能指标ALT、AST、LDH水平和肝组织Hyp活性显著降低，以及肝组织中MDA和SOD活性明显增加，血清炎性因子TNF-α、IL-6和IL-1β水平显著降低，肝组织的病理损伤以及肝细胞凋亡情况得到明显改善，肝组织中SIRT1蛋白表达增加，HMGB1和NF-κB蛋白的表达降低(P＜0。05或P＜0。01)。结论 丹参绞股蓝茶能有效保护急性肝损伤，其作用机制可能与调控SIRT1/HMGB1/NF-κB信号通路，减轻肝细胞炎性凋亡有关。
Protective effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on apoptosis of mice cells with acute liver injury induced by carbon tetrachloride
Objective Use of silencing information regulatory factor 1(SIRT1)/high mobility group protein B1(HMGB1)/nuclear transcription factor-KB(NF-κB)to investigate the inhibitory effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on hepatocyte inflammatory apoptosis in mice with carbon tetrachloride(CC14)-induced acute liver injury.Methods C57BL/6 mice were randomly divided into a control group;model group;resveratrol group;Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea low-,medium-,and high-dose groups;and a positive drug group.The mice were given continuous intragastric administration of the treatments for 14 days.An acute liver injury model was established by intraperitoneal injection of 0.5％CC14 olive oil solution(5 mL/kg).The levels of alanine transferase(ALT),aspartate transferase(AST),lactate dehydrogenase(LDH)in serum and hydroxyproline(Hyp),malondialdehyde(MDA),and superoxide dismutase(SOD)in the liver were determined by biochemical method.Serum levels of inflammatory tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1 β were determined by enzyme-linked immunosorbent assay.Eosin-hematoxylin and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling(TUNEL)staining were used to examine the pathological morphology and apoptosis in liver tissues.The protein expression of SIRT1,HMGB1,and NF-κB were detected by Western Blot.Results Compared with those of the control group,the model group's serum ALT,AST,and LDH levels and liver tissue Hyp activity were significantly increased;MDA and SOD activities in liver tissue were significantly decreased;the levels of inflammatory factors TNF-α,IL-6,and IL-1 β in liver tissue were significantly increased;and there were obvious signs of pathological injury and hepatocyte apoptosis in the liver tissue.The expression of SIRT1 protein decreased significantly,while the expression of HMGB1 and NF-κB proteins increased,in liver tissue.Compared with those of the model group,the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high group and resveratrol group serum levels of ALT,AST,and LDH and liver tissue Hyp activity were significantly decreased;MDA and SOD activities in liver tissue were significantly increased;the levels of serum inflammatory factors TNF-α,IL-6,and IL-1β were decreased;and pathological injury to liver tissue and the apoptosis of liver cells were significantly improved.The expression of SIRT1 protein in liver tissue was increased,while the expression of HMGB1 and NF-κB protein were decreased in the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high-dose group and resveratrol group(P＜0.05 or P＜0.01).Conclusions Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea effectively protects against acute liver injury,and its mechanisms may be related to the regulation of the SIRT1/HMGB1/NF-κB signaling pathway and the alleviation hepatocyte inflammatory apoptosis.
acute liver injuryinflammatory apoptosissilent information regulator 1high mobility group protein B1nuclear transcription factor-KB
万方数据
维普
