1. 河北省保定市传染病医院肝五科,河北,保定,071000
2. 河北省保定市传染病医院结核二科,河北,保定,071000
3. 河北省保定市传染病医院肝六科,河北,保定,071000
4. 河北省保定市传染病医院肝四科,河北保定,071000
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李木松, 张贵贤, 陈晖, 等. 茵陈蒿汤对酒精性肝纤维化模型大鼠内质网应激介导肝细胞凋亡相关分子的影响[J]. 上海中医药杂志, 2019,53(6):75-79.
LI Musong, ZHANG Guixian, CHEN Hui, et al. Effects of Yinchenhao Decoction on molecules associated with hepatocyte apoptosis mediated by endoplasmic reticulum stress in rats with alcoholic liver fibrosis [J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):75-79.
李木松, 张贵贤, 陈晖, 等. 茵陈蒿汤对酒精性肝纤维化模型大鼠内质网应激介导肝细胞凋亡相关分子的影响[J]. 上海中医药杂志, 2019,53(6):75-79. DOI: 10.16305/j.1007-1334.2019.06.019.
LI Musong, ZHANG Guixian, CHEN Hui, et al. Effects of Yinchenhao Decoction on molecules associated with hepatocyte apoptosis mediated by endoplasmic reticulum stress in rats with alcoholic liver fibrosis [J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):75-79. DOI: 10.16305/j.1007-1334.2019.06.019.
目的:观察茵陈蒿汤对酒精性肝纤维化大鼠肝组织内质网应激凋亡通路的影响,探讨茵陈蒿汤抗酒精性肝纤维化的作用机制。 方法:将40只SD大鼠采用白酒-玉米油-吡唑混合液灌胃联合小剂量CCl,4,腹腔注射复制大鼠酒精性肝纤维化模型,8周后随机分为对照组、模型组和茵陈蒿汤低剂量组、茵陈蒿汤高剂量组,每组10只。模型组以白酒混合物低剂量(6 mL/kg)持续灌胃,茵陈蒿汤低剂量组、高剂量组分别给予茵陈蒿汤1.75 g/kg(5 mL/kg)、3.5 g/kg(10 mL/kg),对照组灌胃给予等量的0.9%NaCl溶液。4周后检测大鼠肝指数及血清 AST、ALT 水平;采用流式细胞学技术检测各组大鼠肝细胞悬液凋亡率的比例;采用Western blot 法检测各组大鼠肝脏GRP78、Cleaved-Caspase-12和Cleaved-Caspase-3蛋白表达情况。 结果:①与对照组比较,模型组大鼠肝指数水平显著升高(P,<,0.05);与模型组比较,茵陈蒿汤各剂量组大鼠肝指数水平显著降低(P,<,0.05);与茵陈蒿汤低剂量组比较,茵陈蒿汤高剂量组大鼠肝指数水平显著降低(P,<,0.05)。②与对照组比较,模型组大鼠血清中ALT、AST水平显著升高(P,<,0.05);与模型组比较,茵陈蒿汤各剂量组ALT、AST水平显著降低(P,<,0.05);与茵陈蒿汤低剂量组比较,茵陈蒿汤高剂量组大鼠血清中ALT、AST水平显著降低(P,<,0.05)。③与对照组比较,模型组大鼠肝细胞晚期凋亡率及总凋亡率显著升高(P,<,0.05);与模型组比较,茵陈蒿汤各剂量组晚期凋亡率及总凋亡率显著降低(P,<,0.05);与茵陈蒿汤低剂量组比较,茵陈蒿汤高剂量组大鼠肝细胞早期、晚期及总凋亡率均明显降低(P,<,0.05)。④与对照组比较,模型组大鼠肝组织Cleaved-Caspase-3蛋白表达水平显著上调(P,<,0.05);与模型组比较,茵陈蒿汤各剂量组大鼠肝组织GRP78、Cleaved-Caspase-12和 Cleaved-Caspase-3蛋白表达水平显著下调(P,<,0.05);与茵陈蒿汤低剂量组比较,茵陈蒿汤高剂量组大鼠肝组织GRP78和Cleaved-Caspase-12蛋白表达水平显著下调(P,<,0.05)。 结论:茵陈蒿汤可明显改善酒精性肝纤维化大鼠的肝组织损伤,抑制内质网应激介导的肝细胞凋亡。
Objective:To observe the effects of Yinchenhao Decoction on the endoplasmic reticulum stress apoptosis pathway in rats with alcoholic liver fibrosis, and to explore the mechanism of Yinchenhao Decoction against alcoholic liver fibrosis. Methods40 SD rats were given liquor-corn oil-pyrazole mixture by gavage and low-dose CCl,4, by intraperitoneal injection to replicate the rat model of alcoholic liver fibrosis. After 8 weeks, the model animals were randomly assigned into blank group, model group and low- and high-dose Yinchenhao Decoction groups, 10 rats in each group. The model group was given continuously with low dose of liquor mixture (6 mL/kg), Yinchenhao Decoction was given at 1.75 g/kg (5 mL/kg) and 3.5 g/kg (10 mL/kg) respectively in the low- and high-dose groups. The control group was given 0.9% NaCl solution of equal volume by gavage. 4 weeks later, the liver index and the levels of serum AST and ALT were determined; the proportion of hepatocyte suspension apoptosis rate in each Yinchenhao Dcoction group was determined by flow cytometry; Western blot was used to detect the expression of GRP78, Cleaved-Caspase-12 and Cleaved-Caspase-3 in liver of rats in each group. Results:①Compared with the control group, the liver index increased significantly in the model group (P,<,0.05); compared to the model group, the liver index decreased significantly in the Yinchenhao groups (P,<,0.05); compared with the low-dose Yinchenhao group, the index decreased significantly in the high-dose Yinchenhao group (P,<,0.05). ②Compared to the control group, the levels of ALT and AST in serum increased significantly in the model group (P,<,0.05); compared to the model group, the levels of ALT and AST in serum decreased significantly in the Yinchenhao groups (P,<,0.05); compared with the low-dose Yinchenhao group, the levels decreased significantly in the high-dose Yinchenhao group (P,<,0.05). ③Compared to the control group, the late and total apoptosis rates increased significantly in the model group (P,<,0.05); compared to the model group, the late and total apoptosis rates decreased significantly in the Yinchenhao groups (P,<,0.05); compared with the low-dose Yinchenhao group, the early, late and total apoptosis rates decreased significantly in the high-dose Yinchenhao group (P,<,0.05). ④Compared to the control group, the expression level of Cleaved-Caspase-3 was upregulated significantly in the model group (P,<,0.05); compared to the model group, the expression levels of GRP78, Cleaved-Caspase-12 and Cleaved-Caspase-3 were downregulated significantly in the Yinchenhao groups (P,<,0.05); compared with the low-dose Yinchenhao group, the expression levels of GRP78 and Cleaved-Caspase-12 were downregulated significantly in the high-dose Yinchenhao group (P,<,0.05). Conclusion:Yinchenhao Decoction can significantly improve liver tissue damage in rats with alcoholic liver fibrosis and inhibit hepatocyte apoptosis mediated by endoplasmic reticulum stress.
茵陈蒿汤酒精性肝纤维化大鼠内质网应激细胞凋亡
Yinchenhao Decoctionalcoholic liver fibrosisratsendoplasmic reticulum stresscell apoptosis
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