复方灵丹胶囊对肝纤维化小鼠模型肝功能及纤维化程度的影响

涂芷馨; 王妍; 黄剑洁; 金清龙

doi:10.3969/j.issn.1001-5256.2023.04.010

复方灵丹胶囊对肝纤维化小鼠模型肝功能及纤维化程度的影响

DOI: 10.3969/j.issn.1001-5256.2023.04.010

吉林大学第一医院肝胆胰内科，长春 130000

基金项目:

国家自然科学基金 (81270484)

伦理学声明：本研究方案于2019年6月1日经由吉林大学实验动物伦理委员会审批，批号：SY201906010，符合实验室动物管理与使用准则。

利益冲突声明：本研究不存在研究者、伦理委员会成员以及与公开研究成果有关的利益冲突。

作者贡献声明：涂芷馨负责课题设计，资料分析，撰写论文；王妍、黄剑洁参与收集数据，修改论文；金清龙负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
金清龙，jinql@jlu.edu.cn (ORCID: 0000-0002-4626-9828)

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出版历程
- 收稿日期: 2022-09-07
- 录用日期: 2022-10-24
- 出版日期: 2023-04-20

Effect of Compound Lingdan Capsule on liver function and fibrosis degree in mice with liver fibrosis

Department of Hepatology, The First Hospital of Jilin University, Changchun 130000, China

Research funding:

National Natural Science Foundation of China (81270484)

More Information

Corresponding author: JIN Qinglong, jinql@jlu.edu.cn(ORCID: 0000-0002-4626-9828)

摘要

摘要: 目的研究复方灵丹胶囊对肝纤维化小鼠模型血清生化指标及肝脏纤维化程度的影响。方法 125只SPF级雄性C57BL/6小鼠随机分为正常对照组(n=5)；CCl₄模型组(n=15), CCl₄造模低、中、高剂量给药组(0.8、1.6、3.2 mg·g^-1·d^-1)(每组各15只)；DDC模型组(n=15)，DDC造模低、中、高剂量给药组(0.8、1.6、3.2 mg·g^-1·d^-1) (每组各15只)。造模成功后，给药组予以相应剂量复方灵丹胶囊混悬液灌胃，正常对照组及模型组以等量生理盐水灌胃，连续给药4周，眼球取血，留取血清，检测AST、ALT、白蛋白、胆红素，留取肝右叶同一部位组织进行病理观察及天狼星红、α1-Ⅰ型胶原蛋白抗体及α-平滑肌肌动蛋白抗体染色。计量资料多组间比较采用单因素方差分析，进一步两两比较采用Bonferroni法。结果与模型组对比，复方灵丹胶囊给药后，各剂量组小鼠血清ALT明显下降、白蛋白升高(P值均＜0.05)，中、高剂量给药组血清AST、胆红素水平均降低(P值均＜0.05)，且各指标高剂量组较低剂量组差异更显著(P值均＜0.05)。各剂量组小鼠肝组织病理变化均得到不同程度的改善，天狼星红染色阳性细胞显著减少，α-平滑肌肌动蛋白、α1-Ⅰ型胶原蛋白表达出现不同程度的下降。结论复方灵丹胶囊能够改善肝纤维化小鼠肝功能、降低其肝纤维化程度。
- 肝硬化 /
- 小鼠, 近交C57BL /
- 复方灵丹胶囊
Abstract: Objective To investigate the effect of Compound Lingdan Capsule on serum biochemical parameters and liver fibrosis degree in a mouse model of liver fibrosis. Methods A total of 125 specific pathogen-free male C57BL/6 mice were randomly divided into normal control group with 5 mice, CCl₄ model group with 15 mice, low-, middle-, and high-dose CCl₄ groups (0.8, 1.6, and 3.2 mg·g^-1·d^-1) with 15 mice in each group, DDC model group with 15 mice, and low-, middle-, and high-dose DDC groups (0.8, 1.6, and 3.2 mg·g^-1·d^-1) with 15 mice in each group. After successful modeling, the mice in the administration groups were given Compound Lingdan Capsule suspension at the respective doses by gavage, and those in the normal control group and the model group were given an equal volume of normal saline by gavage, for 4 consecutive weeks. Blood samples were collected from the eyeballs, and serum was used to measure aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, and bilirubin. Liver tissue samples were collected at the same site of the right lobe of the liver for pathological observation, Sirius Red staining, α-SMA antibody staining, and COL1A1 antibody staining. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Bonferroni method was used for further comparison between two groups. Results Compared with the model group, each dose group had significant reductions in the serum level of ALT and a significant increase in the serum level of albumin after the administration of Compound Lingdan Capsule (all P < 0.05), the levels of AST and bilirubin in the middle and high dose groups were lower (all P < 0.05), and the difference of each index in the high dose group was more significant than that in the low dose group (all P < 0.05). Each dose group had varying degrees of improvement in the pathological changes of the liver and a significant reduction in the number of Sirius Red staining-positive cells, as well as varying degrees of reduction in the protein expression of α-SMA and COL1A1. Conclusion Compound Lingdan Capsule can improve liver function and reduce liver fibrosis degree in mice with liver fibrosis.
- Liver Cirrhosis /
- Mice, Inbred C57BL /
- Compound Lingdan Capsule

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图 1 各组小鼠肝组织不同染色结果(×100)

Figure 1. Images of different staining of liver tissues(×100)

下载: 全尺寸图片幻灯片

表 1 小鼠分组、造模及给药情况

Table 1. Grouping, modeling and drug administration of mice

分组	动物数(只)	定义
正常对照组	5	给予基础饲料喂养。等量生理盐水灌胃，每日给药，连续4周
CCl₄造模组		基础饲料喂养的基础上予以CCl₄溶液腹腔注射。以5 mL/kg剂量皮下注射体积分数为20%的CCl₄油剂溶液(CCl₄∶橄榄油=1∶4)，每3 d 1次，连续6周
CCl₄模型组	15	等量生理盐水灌胃，每日给药，连续4周
CCl₄-L组	15	复方灵丹胶囊低剂量0.8 mg·g^-1·d^-1
CCl₄-M组	15	复方灵丹胶囊中剂量1.6 mg·g^-1·d^-1
CCl₄-H组	15	复方灵丹胶囊高剂量3.2 mg·g^-1·d^-1
DDC造模组		以0.1% DDC饲料喂养，连续6周
DDC模型组	15	等量生理盐水灌胃，每日给药，连续4周
DDC-L组	15	复方灵丹胶囊低剂量0.8 mg·g^-1·d^-1
DDC-M组	15	复方灵丹胶囊中剂量1.6 mg·g^-1·d^-1
DDC-H组	15	复方灵丹胶囊高剂量3.2 mg·g^-1·d^-1

下载: 导出CSV

表 2 CCl₄各组小鼠血清ALT、AST、胆红素和白蛋白水平比较

Table 2. Comparison of serum ALT, AST, bilirubin and albumin levels of mice in each group of CCl₄

组别	动物数 (只)	ALT (U/L)	AST (U/L)	胆红素 (μmol/L)	白蛋白 (ng/mL)
正常对照组	3	39.32±4.68¹⁾	34.53±8.47¹⁾	5.69±2.08¹⁾	399.26±39.89¹⁾
CCl₄模型组	10	91.40±11.63	89.87±12.77	16.66±2.41	145.14±23.70
CCl₄-L组	15	75.57±11.03¹⁾	77.31±11.28	14.57±2.38	185.51±30.21¹⁾
CCl₄-M组	15	67.04±9.97¹⁾	69.21±13.07¹⁾	13.23±2.23¹⁾	217.04±26.57¹⁾
CCl₄-H组	15	60.37±10.97¹⁾²⁾	62.50±12.05¹⁾²⁾	11.95±2.19¹⁾²⁾	240.33±34.97¹⁾²⁾
F值		20.68	15.72	16.08	47.69
P值		＜0.000 1	＜0.000 1	＜0.000 1	＜0.000 1
注：与CCl₄模型组比较，1)P＜0.05；与CCL₄-L组比较，2)P＜0.05。

下载: 导出CSV

表 3 DDC各组小鼠血清ALT、AST、胆红素和白蛋白水平比较

Table 3. Comparison of serum ALT, AST, bilirubin and albumin levels of mice in each group of DDC

组别	动物数 (只)	ALT (U/L)	AST (U/L)	胆红素 (μmol/L)	白蛋白 (ng/mL)
正常对照组	3	39.32±4.68¹⁾	34.53±8.47¹⁾	5.69±2.08¹⁾	399.26±39.89¹⁾
DDC模型组	10	94.76±17.26	90.72±14.31	16.57±2.25	136.72±33.50
DDC-L组	11	79.48±17.18	77.08±14.04	14.45±2.28	199.60±51.22
DDC-M组	11	71.36±17.09¹⁾	69.19±14.68¹⁾	13.08±2.23¹⁾	226.35±55.53¹⁾
DDC-H组	11	62.98±18.64¹⁾	61.97±13.64¹⁾	11.79±2.31¹⁾	247.61±62.51¹⁾
F值		8.20	11.82	15.68	16.74
P值		＜0.000 1	＜0.000 1	＜0.000 1	＜0.000 1
注：与DDC模型组比较，1)P＜0.05。

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