邢安丽;赵鲲鹏;张秋菊;李婕妠;陈世玉;郭嘉琪;张铭.舒肝化癥方对肝纤维化模型大鼠VEGF、CTGF、CD31及vWF表达的影响及其相关性研究[J].中医药信息,2024,41(6):19-27 |
舒肝化癥方对肝纤维化模型大鼠VEGF、CTGF、CD31及vWF表达的影响及其相关性研究 |
Effect and Correlation Study of Shugan Huazheng Formula on VEGF, CTGF, CD31, and vWF Expression in Liver Fibrosis Model Rats |
投稿时间:2023-11-06 录用日期:2023-12-01 |
DOI:10.19656/j.cnki.1002-2406.20240604 |
中文关键词: 血管内皮生长因子 结缔组织生长因子 血小板-内皮细胞黏附分子 血管性血友病因子 肝纤维化 舒肝化癥方 |
英文关键词: VEGF CTGF CD31 vWF Hepatic fibrosis Shugan Huazheng Formula |
基金项目:基于TGF-β11/smad与JAK/STAT多信号转导通路的舒肝和络醒脾法抗肝纤维化作用机制研究,项目批准号81660772 |
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中文摘要: |
目的:以四氯化碳(CCl4)致大鼠肝纤维化为研究对象,研究舒肝化癥方(SGHZD)对大鼠肝纤维化血瘀证的影响及机理,分析血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)、血小板-内皮细胞黏附分子(CD31)及血管性血友病因子(vWF)在SGHZD抗肝纤维化过程中的表达及其相关性。方法:SPF级SD大鼠,雄性,54只,按随机原则分成空白组、模型组(2.0 mg/kg 40%CCl4橄榄油溶液+等量的0.9%的生理盐水)、阳性对照组(2.0 mg/kg 40%CCl4橄榄油溶液+0.2 mg/kg秋水仙碱溶液)、SGHZD高剂量组(2.0 mg/kg 40%CCl4橄榄油溶液+29.52g/kgSGHZD)、SGHZD中剂量组(2.0 mg/kg 40%CCl4橄榄油溶液+14.76 g/kgSGHZD)和SGHZD低剂量组(2.0 mg/kg 40%CCl4橄榄油溶液+7.38 g/kgSGHZD),各9只。造模时间为8周,每周2次,从造模第2天起,用药治疗每天1次,治疗8周,实验期间动态监测所有大鼠的生理情况,马松染色(Masson)对肝组织病理形态学变化和胶原纤维沉积情况进行观察,实时荧光定量聚合酶链式反应(Real-time PCR)检测VEGF、CTGF、CD31、vWF mRNA的表达水平,Western blot检测VEGF, CTGF,CD31, vWF的蛋白表达。结果:与空白组相比,模型组大鼠整体状况明显下降,Masson染色结果显示大鼠汇管区和中央静脉区形成假小叶,并伴有纤维间隔形成,胶原纤维沉积增多且有显著意义(P <0.05);RT-qPCR、Western blot结果显示,VEGF、CTGF、CD31、vWF mRNA和蛋白表达均上升(P <0.05);与模型组相比,阳性对照组和SGHZD组各剂量Masson染色显示胶原纤维沉积明显减少,纤维间隔程度减轻,差异有统计学意义(P <0.05),RT-qPCR、Western blot结果显示VEGF、CTGF、CD31、vWF mRNA及蛋白的表达量均下降,但以阳性对照组及中药高剂量组效果最好(P均<0.05);将Masson染色胶原容积分数分别与VEGF、CTGF、CD31及vWF蛋白表达进行相关性分析,发现VEGF、CTGF、CD31及vWF在肝纤维化的形成过程中随着肝纤维化程度加重其表达上调,而在SGHZD抗肝纤维化过程中随着胶原容积分数减少其表达减少;对VEGF、CTGF、CD31及vWF蛋白表达进行相关性分析,发现VEGF、CTGF、CD31及vWF在肝纤维化的形成发展中和SGHZD抗肝纤维化的过程中均呈显著正相关。结论:SGHZD对CCl4所致肝纤维化大鼠有显著疗效,其作用机理可能与VEGF、CTGF、CD31及vWF的表达、病理性血管生成、肝窦毛细血管化及肝纤维化血瘀证相关。 |
英文摘要: |
Objective: To study the effect and mechanism of Shugan Huazheng Formula(SGHZD, hereinafter the same) on the blood stasis evidence of hepatic fibrosis in rats with carbon tetrachloride(CCl4)-induced hepatic fibrosis and analyze the expression and correlation of vascular endothelial growth factor(VEGF), connective tissue growth factor(CTGF), platelet-endothelial cell adhesion molecule(CD31), and vascular hemophilic factor(v WF) expression and its correlation in the process of anti-hepatic fibrosis in SGHZD.METHODS: SPF grade SD rats, males, 54 rats, were divided into the following groups according to the principle of randomization: blank group, model group(2. 0 mg/kg 40% CCl4 olive oil solution + equal amount of 0. 9%saline), positive control group(2. 0 mg/kg 40% CCl4 olive oil solution + 0. 2 mg/kg colchicine solution), and SGHZD high-dose group(2. 0 mg/kg 40% CCl4 olive oil solution + 29. 52 g/kg SGHZD), SGHZD medium-dose group(2. 0 mg/kg 40% CCl4 olive oil solution + 14. 76 g/kg SGHZD), and SGHZD low-dose group(2. 0 mg/kg 40% CCl4 olive oil solution + 7. 38 g/kg SGHZD), each of which consisted of 9 animals. The modeling time was 8 weeks, twice a week, from the 2nd day of modeling, treatment with the drug was once a day for 8 weeks, the physiological conditions of all rats were dynamically monitored during the experimental period, Masson staining(Masson) was performed for the observation of pathomorphological changes in liver histology and collagen fiber deposition, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed for the detection of VEGF, CTGF, VEGF, CTGF, CD31, vWF mRNA expression levels were detected by real-time fluorescence quantitative polymerase chain reaction(Realtime PCR), and the protein expression of VEGF, CTGF, CD31, vWF was detected by western blot.RESULTS: Compared with the blank group, the overall condition of the rats in the model group was significantly decreased. Masson staining results showed that pseudofollicles were formed in the confluent area and central venous area of the rats, accompanied by the formation of fibrous septa, and the deposition of collagen fibers was increased and significant(P < 0. 05); RT-qPCR and western blot results showed that the expression levels of VEGF, CTGF, CD31, and vWF mRNAs in the model group were significantly decreased, CTGF, CD31, vWF mRNA and protein expression were increased(P < 0. 05); compared with the model group, Masson staining of each dose in the positive control group and the SGHZD group showed a significant decrease in collagen fiber deposition and a reduction in the degree of fibrous septum, and the difference was statistically significant(P < 0. 05), and the RT-qPCR, Western The results of RT-qPCR, Western blot showed that the expression of VEGF, CTGF, CD31, and vWF mRNA and protein decreased, but the positive control group and the high-dose group of traditional Chinese medicine had the best effect(both P < 0. 05); Masson staining of collagen volume fraction was correlated with the protein expression of VEGF, CTGF, CD31, and vWF, respectively, and found that VEGF, CTGF, CD31, and vWF were found in the liver, CD31and vWF were correlated with VEGF, CTGF, CD31 and vWF protein expression, and found that VEGF, CTGF, CD31 and vWF were up-regulated with the increase in the degree of hepatic fibrosis during the development of hepatic fibrosis and with the decrease in the collagen volume fraction during the anti-hepatic fibrosis process of SGHZD. SGHZD anti-hepatic fibrosis were all significantly positively correlated.CONCLUSION: SGHZD has significant efficacy in CCl4-induced hepatic fibrosis in rats, and its mechanism of action may be related to the expression of VEGF, CTGF, CD31 and vWF, pathologic angiogenesis, hepatic sinusoidal capillarization, and blood stasis evidence of hepatic fibrosis. |
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