中国中药杂志

2020, v.45(21) 5256-5264

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兰坪虫草多糖组分分析及缓解小鼠肝纤维化研究
Component analysis of Ophiocordyceps lanpingensis polysaccharides and study on alleviation of hepatic fibrosis in mice by polysaccharides

柯迎妹;江敏;周树波;虞泓;王娟;葛锋;
KE Ying-mei;JIANG Min;ZHOU Shu-bo;YU Hong;WANG Juan;GE Feng;Faculty of Life Science and Technology, Kunming University of Science and Technology;School of Ecology and Environmental Science, Yunnan University;Research Center of Cordyceps Development and Utilization of Kunming;

摘要(Abstract):

兰坪虫草Ophiocordyceps lanpingensis作为云南省少数民族用药,主要用来治疗肝、肾等疾病,但其药理机制尚不明确。该研究分析了兰坪虫草多糖的单糖组分及含量,探讨了兰坪虫草多糖对小鼠肝纤维化的改善作用及机制。结果表明,兰坪虫草多糖主要由甘露糖、葡萄糖、半乳糖和阿拉伯糖组成,质量分数为19.1%,21.8%,21.1%,38.0%。基于四氯化碳诱导的小鼠肝纤维化模型,兰坪虫草多糖可明显缓解小鼠肝组织炎症状态和纤维化程度,逆转因四氯化碳导致的小鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)含量的升高,恢复肝脏正常功能。该研究证明了兰坪虫草多糖可显著降低肝组织中纤维化标志物α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)、Ⅰ型胶原(collagen type 1,Col-1)mRNA表达水平和羟脯氨酸(hydroxyproline,HYP)含量,同时升高肝组织中抗氧化剂超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽(glutathione,GSH)的含量,减少脂质过氧化产物丙二醛(malondialdehyde,MDA)的合成;此外,兰坪虫草多糖还可抑制肝纤维化小鼠肝脏中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和核因子κB(nuclear factor-kappa B,NF-κB)的基因表达水平;进一步研究表明,兰坪虫草多糖降低了肝组织的凋亡指数,下调了肝脏中Bcl-2关联X蛋白(Bcl-2 associated X protein,Bax)、半胱氨酸蛋白酶-3(cysteinyl aspartate specific proteinase-3,caspase-3)和半胱氨酸蛋白酶-9(cysteinyl aspartate specific proteinase-9,caspase-9)蛋白表达水平,促进B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白表达,具有抑制肝细胞凋亡的作用。因此,兰坪虫草多糖对小鼠肝纤维化的改善机制可能是通过缓解机体氧化应激、减轻炎症反应以及抗肝细胞凋亡等多层面共同作用的结果。
Ophiocordyceps lanpingensis is mainly used as an ethnic medicine to treat the diseases of liver, kidney and other diseases, but the pharmacological mechanism is not clear yet. In this study, the components and contents of monosaccharides in the O.lanpingensis polysaccharides(OLP) were analyzed. The results showed that the OLP were composed of mannose, glucose, galactose and arabinose, with mass percentages of 19.1%, 21.8%, 21.1%, and 38.0%, respectively. Based on the hepatic fibrosis model induced by CCl_4 in mice, OLP could significantly relieve the inflammation and fibrosis levels of hepatic tissues, reverse the CCl_4-induced increasing levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in mice serum, and recover the functions of liver to a normal state. This study proved that OLP significantly decreased the mRNA expression levels of fibrotic genes, alpha-smooth muscle actin(α-SMA) and collagen type 1(Col-1), as well as the content of hydroxyproline(HYP) in the liver tissues; meanwhile, the contents of antioxidants superoxide dismutase(SOD) and glutathione(GSH) were enhanced and the production of lipid peroxide malondialdehyde(MDA) was reduced. Moreover, OLP inhibited the gene expression levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and nuclear factor kappa B(NF-κB) in the livers of mice. Further study indicated that OLP could restrain the apoptosis of hepatic cells due to the decrease of the apoptosis index and down-regulations of protein expression levels of Bcl-2 associated X protein(Bax), cysteinyl aspartate specific proteinase-3(caspase-3) and cysteinyl aspartate specific proteinase-9(caspase-9), and the promotion of protein expression level of B-cell lymphoma-2(Bcl-2) in livers. To sum up, the mechanism of OLP for alleviating hepatic fibrosis was likely related to the synergy by remitting the oxidative stress of the body, alleviating inflammatory response, anti-apoptosis of hepatic cells, and so on.

关键词(KeyWords): 兰坪虫草;多糖;肝纤维化;氧化应激;凋亡
Ophiocordyceps lanpingensis;polysaccharides;liver fibrosis;oxidative stress;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 云南省万人计划项目(YNWR-QNBJ-2018-059);; 科技部项目——中央引导地方科技发展专项(KC1610530)

作者(Author): 柯迎妹;江敏;周树波;虞泓;王娟;葛锋;
KE Ying-mei;JIANG Min;ZHOU Shu-bo;YU Hong;WANG Juan;GE Feng;Faculty of Life Science and Technology, Kunming University of Science and Technology;School of Ecology and Environmental Science, Yunnan University;Research Center of Cordyceps Development and Utilization of Kunming;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200628.401

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