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覃妮,黄曙院,周柳,黄华萍,沈新辉,周琳,孙东琪,陆世银.岩黄连总生物碱对四氯化碳诱导肝纤维化大鼠的改善作用研究[J].湖南中医药大学学报,2023,43(12):2191-2198[点击复制] |
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| 岩黄连总生物碱对四氯化碳诱导肝纤维化大鼠的改善作用研究 |
| 覃妮,黄曙院,周柳,黄华萍,沈新辉,周琳,孙东琪,陆世银 |
| (柳州市中医医院(柳州市壮医医院), 中药(壮瑶药)制剂研发重点实验室, 中药(壮瑶药)制剂开发工程技术研究中心, 广西 柳州 545026) |
| 摘要: |
| 目的 研究岩黄连总生物碱对四氯化碳（carbon tetrachloride，CCl4）诱导的肝纤维化大鼠的保护作用及机制。方法 通过向SD大鼠腹腔给予30% CCl4橄榄油溶液（0.1 mL/100 g）诱导复制肝纤维化模型，2次/周，连续12周。将肝纤维化大鼠随机分为模型组，岩黄连总生物碱低、中、高剂量（25、50、100 mg/kg）组及联苯双酯（30 mg/kg）组，另设正常组大鼠，各组均为8只。各给药组于造模第11周按设定剂量灌胃相应药物（1.0 mL/100 g）干预，1次/d，连续2周。正常组与模型组大鼠每天灌胃等体积的生理盐水。末次给药24 h后，收集大鼠血清、肝脏及脾脏组织，测定大鼠肝脾指数；采用HE染色和Masson染色观察肝组织病理学变化。比色法检测各组血清丙氨酸转氨酶（alanine aminotransferase，ALT）、天冬氨酸转氨酶（aspartate aminotransferase，AST）、总蛋白（total protein，TP）、白蛋白（albumin，ALB）、丙二醛（malondialdehyde，MDA）、超氧化物歧化酶（superoxide dismutase，SOD）与谷胱甘肽过氧化物酶（Glutathione peroxidase，GSH-Px）的含量；通过ELISA检测血清透明质酸（hyaluronic acid，HA）、层粘连蛋白（laminin，LN）、Ⅲ型前胶原（typeⅢprocollagen，PCⅢ）及Ⅳ型胶原（collagenⅣ，Ⅳ-C）、转化生长因子-β1（transforming growth factor-β1，TGF-β1）、白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-8（interleukin-8，IL-8）及白细胞介素-6（interleukin-6，IL-6）表达水平。结果 与正常组相比，模型组大鼠反应迟钝，肝脾指数增大（P<0.01），肝组织呈现明显纤维化，血清ALT、AST、MDA、LN、HA、PCⅢ和Ⅳ-C水平显著升高（P<0.01），血清TP、ALB、SOD及GSH-Px水平显著下降（P<0.01）；血清中TGF-β1、IL-1β、IL-8及IL-6蛋白表达水平显著上调（P<0.01）。与模型组比较，岩黄连总生物碱中、高剂量组均能明显改善大鼠状态和肝脾指数（P<0.05，P<0.01），有效调节大鼠肝组织病理学损伤和纤维增生；显著降低大鼠血清ALT、AST、MDA含量（P<0.01），明显提高血清TP、ALB、SOD及GSH-Px的表达水平（P<0.05，P<0.01）；并能明显下调LN、HA、PCⅢ、Ⅳ-C、TGF-β1、IL-1β、IL-8及IL-6蛋白的表达水平（P<0.05，P<0.01）。结论 岩黄连总生物碱具有明显的抗肝纤维化作用，其作用机制可能与抗氧化、抑制炎症因子表达相关。 |
| 关键词: 岩黄连 总生物碱 四氯化碳 肝纤维化 抗氧化 炎症因子 |
| DOI：10.3969/j.issn.1674-070X.2023.12.008 |
| 投稿时间：2023-05-08 |
| 基金项目:柳州市科技攻关与新产品试制（2020NBAA0802）；广西中医药大学自然科学基金面上项目（2020MS055）。 |
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| Improvement effects of total alkaloids from Yanhuanglian (Corydalis saxicola Bunting) on carbon tetrachloride-induced liver fibrosis in rats |
| QIN Ni,HUANG Shuyuan,ZHOU Liu,HUANG Huaping,SHEN Xinhui,ZHOU Lin,SUN Dongqi,LU Shiyin |
| (Liuzhou Key Laboratory of Preparation Research and Development of Chinese Medicines (Zhuang and Yao), Liuzhou Engineering Research Center for Preparation Technology of Chinese Medicines (Zhuang and Yao), Liuzhou Chinese Medical Hospital, Liuzhou, Guangxi 545026, China) |
| Abstract: |
| Objective To investigate the protective effects and mechanism of total alkaloids of Yanhuanglian(Corydalis saxicola Bunting)(TACS) on carbon tetrachloride-induced liver fibrosis in rats.Methods The SD rats were injected intraperitoneally with 30% carbon tetrachloride(CCl4) olive oil solution(0.1 mL/100 g) to establish liver fibrosis model, twice a week for 12 consecutive weeks. Liver fibrosis rats were randomly divided into five groups with eight animals per group:model group, low-, medium-, and high-dose TACS(25, 50, 100 mg/kg) groups, and bifendate group(30 mg/kg). Additionally, a normal group(8 rats) was also set up.The normal group and model group were given constant volume of saline intragastrically, while the other groups were intervened by intragastric administration of the corresponding drug(1.0 mL/100 g) according to the set dose at the 11th week of modeling, once a day for two weeks. After 24 h of the last administration, the serum, liver and spleen tissues were collected to determine the liver and spleen indexes of rats. The pathological changes of liver tissues were observed via hematoxylin-eosin(HE) staining and Masson staining. Colorimetric method was used to check the content of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total protein(TP), albumin(ALB), malondialdehyde(MDA), superoxide dismutase(SOD), and Glutathione peroxidase(GSH-Px) in serum.The expression levels of serum hyaluronic acid(HA), laminin(LN), type Ⅲ procollagen(PCⅢ), collagen Ⅳ(Ⅳ-C), transforming growth factor-β1(TGF-β1), interleukin-1β(IL-1β), interleukin-8(IL-8), and interleukin-6(IL-6) in serum were measured by ELISA.Results Compared with the normal group, the rats in the model group were unresponsive, the liver and spleen indexes increased(P<0.01), the liver tissues showed obvious fibrosis, the levels of ALT, AST, MDA, HA, LN, PCⅢ, and Ⅳ-C increased significantly(P<0.01), the levels of TP, ALB, SOD, and GSH-Px in serum decreased significantly(P<0.01), and the expression levels of TGF-β1, IL-1β, IL-8, and IL-6 in serum were significantly up-regulated(P<0.01). Compared with the model group, the state and liver and spleen indexes of rats in medium-and high-dose TACS groups significantly improved(P<0.05, P<0.01), the pathological damage and fibrous hyperplasia of liver tissues were effectively regulated, the content of ALT, AST, and MDA in serum significantly decreased(P<0.01), the expression levels of TP, ALB, SOD, and GSH-Px significantly increased(P<0.05, P<0.01) and the expression levels of LN, HA, PCⅢ, Ⅳ-C, TGF-β1, IL-1β, IL-8, and IL-6 proteins significantly down-regulated(P<0.05, P<0.01).Conclusion TACS has obvious anti-hepatic fibrosis effects, and its mechanism of action may be related to antioxidation and inhibition of inflammatory factor expression. |
| Key words: Corydalis saxicola Bunting total alkaloid carbon tetrachloride carbon tetrachloride liver fibrosis antioxidation inflammatory factor |
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