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纸质出版日期:2013
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扈晓宇, 张扬. 清热化瘀中药复方对急性肝衰竭大鼠肝组织HMGB1表达的干预研究[J]. 中国实验方剂学杂志, 2013,19(2):172-177.
HU Xiao-yu, ZHANG Yang. Effect of Qingre Huayu Decoction on Expression of HMGB1 in Liver Tissue of Acute Liver Failure Rats[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(2): 172-177.
目的: 观察清热化瘀中药复方——清肝方对急性肝衰竭(acute liver failure
ALF)大鼠高迁移率族蛋白 B1(HMGB1)表达的影响
从mRNA水平探讨其抗ALF的机制。 方法: 采用D-氨基半乳糖(D-GalN)单次腹腔注射构建ALF大鼠模型
125只SD大鼠以是否接受造模和药物干预随机分为空白组、模型组、复方甘草酸苷组和清肝方组
每组再以36
96 h 2个时间点继续随机分为1
2两个亚组
共8组
其中亚组1用于造模后36 h取血及肝组织标本
亚组2用于96 h后观察大鼠的生存率。用全自动生化分析法检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆红素(TBiL);全自动血凝分析法检测血浆凝血酶原时间(PT);常规HE染色作肝组织病理学观察。实时荧光定量(RT-PCR)检测HMGB1 mRNA的表达变化
以管家基因β-actin为对照
2-ΔΔCt法计算目的基因相对表达量。 结果: 模型组、复方甘草酸苷组及清肝方组估计平均生存时间分别为64.6
71.9
83.3 h;log-rank检验提示清肝方组累积生存率高于模型组(P<0.05)。空白组、清肝方组和复方甘草酸苷组肝组织HMGB1 RNA/β-actin值均显著降低(0.006±0.003
0.067±0.033
0.112±0.027 vs 0.245±0.153
均P<0.01)
清肝方组低于复方甘草酸苷组(P<0.01)。与模型组相比
空白组、清肝方组和复方甘草酸苷组在血清ALT
AST
TBiL和血浆PT水平方面与模型组相比均明显下降(P<0.01)
清肝方组低于复方甘草酸苷组(P<0.01)。清肝方组和复方甘草酸苷组肝组织损害程度积分与模型组相比明显下降(1.84±0.13
2.85±0.20 vs 3.56±0.24
均P<0.01)
清热化瘀复方组低于复方甘草酸苷组(P<0.01)。 结论: 清肝方可有效改善D-GalN诱导的大鼠的肝功能、凝血功能及肝脏病理
降低死亡率
其发挥作用的分子机制与抑制HMBG1的表达有关。
Objective: To observe the effect of Qingre Huayu decoction(Qinggan decoction) on expression of high mobility group box 1 protein(HMGB1) in liver tissue of acute liver failure(ALF) rats
and to explore the mechanism of anti-ALF from mRNA level. Method: Rats were injected intraperitoneally with D-GalN 1.4 g· kg-1 to induce ALF rat model. One hundred and twenty five SD rats were randomly divided into four groups
that was normal group
model group
Stronger Neo-Minophagen C (SNMC) group and Qinggan decoction group according to whether or not to accept the modeling and drug intervention. Each group was further randomly divided into two subgroups according to different time points
a total of 8 groups. The first subgroup was used to collect blood and liver tissue samples at hour 36
while the second was applied to observe the survival rate of rats at hour 96.Serum alanine aminotransferase(ALT)
aspartate aminotransferase (AST) and total bilirubin(TBiL) levels were measured via automatic biochemical analysis; plasma prothrombin time(PT) level was measured via automatic coagulation analysis while for liver pathomorphology observation via HE staining. To detect high mobility group box 1 protein(HMGB1) mRNA expression changes via quantitative RT-PCR
and to calculate the target gene relative expression level of the housekeeping gene β-actin as control by 2-ΔΔCt method. Result: After 96-hour treatment
mean survival times of the three groups were 64.6
71.9
83.3 hours respectively; cumulative survival rate of rats in Qinggan decoction group was higher than that of rats in model group(P<0.05). Compared with the model group
the levels of HMGB1 mRNA/β-actin were remarkably reduced in the normal group
Qinggan decoction group and SNMC group(0.006±0.003
0.067±0.033
0.112±0.027 vs 0.245±0.153
all P<0.01). The level of ALT
AST
TBiL and PT were significantly lower in the Qinggan decoction group than in the SNMC group(P<0.01). Compared with the model group
the levels of ALT
AST
TBiL and PT were remarkably reduced in the normal group
Qinggan decoction group and SNMC group(P<0.01). The level of ALT
AST
TBiL and PT were significantly lower in the Qinggan decoction group than in the SNMC group(P<0.01). Compared with the model group
the level of liver tissue damage degree score was also significantly reduced in the Qinggan decoction group and SNMC group(1.84±0.13
2.85±0.20 vs 3.56±0.24
both P<0.01). The level of liver tissue damage degree score was significantly lower in the Qinggan group than in the SNMC group(P<0.01). Conclusion: Qinggan decoction can be effective in improving liver function
blood coagulation
liver pathology and reducing mortality in D-GalN-induced acute liver failure rats
and its molecular mechanisms may be related to inhibition of HMBG1 expression.
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