| 朱芳红,王 文,刘晓姿,苗鹏坤,张晓菊.7种茵陈单体化合物对黄疸大鼠血清培养的hiHep细胞中水通道蛋白8表达的影响[J].,2023,(12):2209-2214 |
| 7种茵陈单体化合物对黄疸大鼠血清培养的hiHep细胞中水通道蛋白8表达的影响 |
| Effects of Seven Monomers from Artemisia capillaris Thunb. on the Expression of Aquaporin 8 in hiHep Cells Cultured in Serum of Jaundiced Rats |
| 投稿时间：2022-11-27 修订日期：2022-12-23 |
| DOI：10.13241/j.cnki.pmb.2023.12.002 |
| 中文关键词: 黄疸 茵陈 水通道蛋白8 单体化合物 |
| 英文关键词: Jaundice Artemisia capillaris Thunb. Aquaporin 8 Monomer compound |
| 基金项目:国家自然科学基金项目（82104718）；空军军医大学第一附属医院2019年度学科助推计划项目（XJZT19MJ45） |
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| 中文摘要: |
| 摘要 目的：探究具有保肝利胆作用的7种茵陈单体化合物（6,7-二甲氧基香豆素、对羟基苯乙酮、绿原酸、咖啡酸、茵陈色原酮、东莨菪内酯和茵陈黄酮）对黄疸大鼠血清培养的hiHep细胞中水通道蛋白8（AQP8）表达的影响。方法：将SD大鼠分为正常组和模型组，通过?琢-萘异硫氰酸酯（ANIT）诱导构建黄疸大鼠模型。检测两组大鼠血清中碱性磷酸酶（ALP）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和总胆红素（TBIL）水平。将人源诱导型肝样细胞（hiHep）随机分为9组：对照组、黄疸血清组（JS组）、黄疸血清+6,7-二甲氧基香豆素组（JS+DME组）、黄疸血清+对羟基苯乙酮组（JS+4-HYD组）、黄疸血清+绿原酸组（JS+CHA组）、黄疸血清+咖啡酸组（JS+CAA组）、黄疸血清+茵陈色原酮组（JS+CAP组）、黄疸血清+东莨菪内酯组（JS+SCO组）和黄疸血清+茵陈黄酮组（JS+ARC组）。对照组细胞使用含10%正常大鼠血清的培养基培养，其他组细胞均使用含10%黄疸大鼠血清的培养基培养。对照组和JS组细胞不做药物处理，其他组细胞均分别使用20 mg/L的相应药物培养48 h。通过四甲基偶氮唑盐（MTT）检测细胞活力。检测各组细胞的蛋白质渗透压反射系数σ。通过qRT-PCR检测细胞中AQP8的mRNA水平，通过Western blot检测细胞中AQP8、环磷酸腺苷（cAMP）和蛋白激酶A（PKA）的蛋白表达水平。结果：与正常组比较，模型组大鼠血清中ALP、ALT、AST和TBIL水平均显著升高（P<0.05）。各组细胞活力差异无统计学意义（F=1.085，P=0.391）。与对照组比较，JS组细胞的σ值降低（P<0.05），与JS组比较，JS+DME组、JS+4-HYD组、JS+CHA组、JS+CAA组、JS+CAP组、JS+SCO组和JS+ARC组细胞的σ值均升高（P<0.05）。与对照组比较，JS组细胞AQP8的mRNA和蛋白相对表达量降低（P<0.05），cAMP和PKA的蛋白相对表达量降低（P<0.05）。与JS组比较，JS+DME组、JS+4-HYD组、JS+CHA组和JS+CAP组细胞AQP8的mRNA和蛋白相对表达量升高（P<0.05），cAMP和PKA的蛋白相对表达量升高（P<0.05）。JS组与JS+CAA组、JS+SCO组和JS+ARC组细胞AQP8的mRNA和蛋白相对表达量以及cAMP和PKA的蛋白相对表达量差异无统计学意义（P>0.05）。结论：本研究表明6,7-二甲氧基香豆素、对羟基苯乙酮、绿原酸和茵陈色原酮可上调黄疸大鼠血清培养的hiHep细胞中AQP8的表达，这可能是茵陈治疗黄疸的机制之一。 |
| 英文摘要: |
| ABSTRACT Objective: To reveal the effects of seven monomers of Artemisia capillaris Thunb. (6,7-dimethoxycoumarin, 4-Hydroxyacetophenone, Chlorogenic acid, Caffeic acid, Capillarisin, Scopoletin, Arcapillin) on the expression of aquaporin 8 (AQP8) in hiHep cells cultured in serum of jaundiced rats. Methods: SD rats were divided into normal group and model group. The jaundice rat model was induced by α-naphthalene isothiocyanate (ANIT). And the serum levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) and total bilirubin (TBIL) in the two groups were detected. Human induced hepatoid cells (hiHep) were randomly divided into 9 groups. They were normal group (Control), jaundice serum group (JS), jaundice serum + 6,7-dimethoxycoumarin group (JS+DME), jaundice serum + 4-Hydroxyacetophenone group (JS+4-HYD), jaundice serum + chlorogenic acid group (JS+CHA), jaundice serum + caffeic acid group (JS+CAA), jaundice serum + capillarisin group (JS+CAP), jaundice serum + scopoletin group (JS+SCO) and jaundice serum + arcapillin group (JS+ARC). The cells in the control group were cultured in the medium containing 10% normal rat serum, and the cells in the other groups were cultured in the medium containing 10% jaundice rat serum. The cells in control group and JS group were not treated with drugs, while the cells in other groups were cultured with corresponding drugs of 20 mg/L for 48 hours. Cell viability was detected by tetrazole salt (MTT). The protein osmotic pressure reflection coefficient σ of each group was detected. The mRNA level of AQP8 was detected by qRT-PCR, and the protein expression of AQP8, cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) were detected by Western blot. Results: Compared with normal group, serum levels of ALP, ALT, AST and TBIL in model group were significantly increased (P<0.05). There was no significant difference in cell viability among all groups (F=1.085, P=0.391). Compared with the control group, the σ value of cell in the JS group was decreased (P<0.05). Compared with JS group, σ value of cell in JS+DME group, JS+4-HYD group, JS+CHA group, JS+CAA group, JS+CAP group, JS+SCO group and JS+ARC group were all increased (P<0.05). Compared with control group, the mRNA and protein relative expression levels of AQP8 in JS group were decreased (P<0.05), and the relative expression levels of cAMP and PKA protein were decreased (P<0.05). Compared with JS group, mRNA and protein relative expressions level of AQP8 in JS+DME group, JS+4-HYD group, JS+CHA group and JS+CAP group were increased (P<0.05), and protein relative expressions of cAMP and PKA group were increased (P<0.05). The relative expression levels of AQP8 mRNA and protein, as well as the relative expression levels of cAMP and PKA protein in JS group and JS+CAA group, JS+SCO group and JS+ARC group were not statistically significant (P>0.05). Conclusion: This study shows that 6,7-dimethoxycoumarin, 4-Hydroxyacetophenone, chlorogenic acid and capillarisin can up-regulate the expression of AQP8 in hiHep cells cultured in serum of jaundice rats, which may be one of the mechanisms for A. capillaris treatment of jaundice. |
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