Effect of Serum Containing Huazhuo Jiedu Recipe on the Oxidative Stress of Cultured Hepatic Stellate Cells

GUO Min; LI Dian-gui

doi:10.11653/syfj2013170258

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Effect of Serum Containing Huazhuo Jiedu Recipe on the Oxidative Stress of Cultured Hepatic Stellate Cells

更新时间：2024-01-04

- Effect of Serum Containing Huazhuo Jiedu Recipe on the Oxidative Stress of Cultured Hepatic Stellate Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 19, Issue 17, Pages: 258-262(2013)
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- DOI：10.11653/syfj2013170258
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- Published：2013
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GUO Min, LI Dian-gui. Effect of Serum Containing Huazhuo Jiedu Recipe on the Oxidative Stress of Cultured Hepatic Stellate Cells[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(17): 258-262.
DOI：

GUO Min, LI Dian-gui. Effect of Serum Containing Huazhuo Jiedu Recipe on the Oxidative Stress of Cultured Hepatic Stellate Cells[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(17): 258-262. DOI： 10.11653/syfj2013170258.

摘要

目的: 研究化浊解毒方含药血清对H2O2诱导的大鼠肝星状细胞(HSC-T6)氧化应激的影响

以探讨化浊解毒方治疗肝纤维化的可能作用机制。方法: SD大鼠ig化浊解毒方不同剂量(0.3

0.6

1.2 g·kg-1)

连续给药10 d

末次给药2 h后取血

制备含药血清;常规培养活化的HSC-T6

采用0.1 mmol·L-1的H2O2制造HSC-T6氧化应激的模型

用不同浓度的含药血清进行干预

分为化浊解毒方组(高、中、低剂量组)、干扰素阳性对照组、模型组及空白对照组(不加任何药物的血清)。干预72 h

放射免疫法检测细胞上清液中超氧化物歧化酶(SOD)

丙二醛(MDA)

还原性谷胱甘肽(GSH)

谷胱甘肽过氧化物酶(GSH-Px)的含量

用RT-PCR及Western blot法检测HSC-T6培养上清液中I型胶原(COL I)及Ⅲ型胶原(COLⅢ)基因及蛋白表达。结果: 与空白对照组比较

经过H2O2处理的HSC-T6 上清液中SOD和GSH-Px的活性明显减低(P<0.05)

MDA和 GSH的含量明显增加(P<0.05);经化浊解毒方含药血清干预72 h后

其高、中剂量组能够提高SOD和GSH-Px的活性(P<0.05)

并降低细胞上清液中MDA和 GSH的含量(P<0.05)

以高剂量组作用明显(P<0.05);与空白对照组比较

HSC-T6 上清液中COL I及COLⅢ基因表达及蛋白含量经过H2O2刺激后明显增加(P<0.05)

经过化浊解毒方干预后

其胶原基因表达及蛋白水平明显降低

尤其高剂量组降低明显(P<0.05)。结论: 化浊解毒方可以减轻因H2O2刺激造成的HSC-T6氧化应激反应

减少氧化应激产物产生

其效果与剂量呈正相关;从基因水平

化浊解毒方能够降低胶原基因的表达

同时抑制其蛋白转录

从而降低细胞外胶原含量

达到抑制肝纤维化的目的。

Abstract

Objective: To observe the inhibitory effect of serum containing Huazhuo Jiedu recipe on oxidative stress in hepatic stellate cell(HSC)-T6 cells. Method: HSC-T6s were divided into control group

model group and serum containing Huazhuo Jiedu recipe group. HSC-T6s in control group were only incubated with high glucose DMEM

HSC-T6s in model group were treated with 0.1 mmol·L-1 H2O2. The methods of thiobarbituric acid

xanthine oxidase and digestion were respectively applied to determine the levels of malondialdehyde(MDA)

superoxide dismutase(SOD)and hydroxyproline(HyP)of culture mediums.The mRNA expression of collagen(COL) I and COLⅢ was determined by RT-PCR and the protein of COL I and COLⅢ was determined by Western blot. Result: Compared with control group

the level of SOD and GSH-Px decreased.the level of MDA and GSH increased significantly in model group(P<0.05). The activity of SOD and GSH-Px increased and the level of MDA and GSH decreased significantly in serum containing Huazhuo Jiedu recipe groups(P<0.05)

especially in the high does and middle does group.The mRNA and the protein expression of COL I and COLⅢ increased in model group

decreased in Huazhuo Jiedu recipe groups(P<0.05). Conclusion: The serum containing Huazhuo Jiedu recipe can dilute the level of oxidation

depress the content of collegen of HSC and lessen fibrosjs.

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