咖啡因治疗大鼠肝纤维化的实验性研究

发布时间：2018-06-26 05:33

本文选题：咖啡因 + 肝纤维化　；参考：《中南大学》2014年硕士论文

【摘要】：目的：利用咖啡因干预四氯化碳(Carbon tetrachloride, CCl4)诱导SD大鼠所形成的肝纤维化,并观察大鼠相关指标的变化,并探讨咖啡因可能的治疗机制。方法：把SD大鼠100只随机分为4组,每组25只,并予橄榄油3ml/(kg大鼠体重)皮下注射空白对照组(n=25),每周2次,共注射8周；将含50%CC14的橄榄油混合溶液以3ml/(kg大鼠体重)皮下注射于模型组(n=25)及咖啡因治疗组(n=25)予以,每周2次,共注射8周；咖啡因对照组(n=25)不予以肝纤维化造模处理。同时,咖啡因治疗组和咖啡因对照组均于第一周开始予以咖啡因灌胃,50mg/(kg大鼠体重),1次/日；以羧甲基纤维素钠(Sodium carboxymethyl cellulose, CMC-Na)用于空白对照组和模型组的大鼠进行灌胃治疗,方法同咖啡因,共治疗8周。在第九周时,大鼠被麻醉后将其处死并称量,分别取肝脏相同部位组织分别进行HE染色和Masson染色并进行评分,取其肝组织进行HE、Masson染色,进行肝脏炎症活动度半定量评分和肝组织纤维化半定量评分,采用免疫组织化学染色后检测肝组织α-平滑肌肌动蛋白(Alpha-Smooth muscle actin, α-SMA)的表达量,并采用实时荧光定量聚合酶链反应(Real time quantitative Polymerase Chain Reaction, Real time qPCR)检测转化生长因子-β1(Transforming growth factor-beta1, TGF-β1)、结缔组织生长因子(Connective tissue growth factor, CTGF)和α-SMA的表达量。结果：与空白对照组相比较,大鼠(模型组)的体重变化、病理评分、TGF-β1、CTGF和α-SMA的表达有明显的增高(P0.01)；与模型组相比较,大鼠(咖啡因治疗组)体重变化、肝纤维化病理评分、TGF-β1、CTGF的表达有明显的下降(p0.01),而α-SMA的表达与TGF-β1、CTGF的表达相比较,前者的降低程度相对不明显(p0.05),但是仍然具有统计学意义；将咖啡因对照组与空白对照组进行比对时,上述检测数据均无明显统计学差异,即p0.05。结论：(1)咖啡因能在一定程度上抑制由CC14诱导的SD大鼠所形成的实验性肝纤维化；(2)在短期内(8周)持续摄入咖啡因后,健康大鼠的肝脏未见显著性改变(P0.05)；(3)咖啡因的抗肝纤维化机制可能为：抑制肝脏炎症反应,抑制HSCs及MFs的活化(a-SMA表达下调),减少促肝纤维化因子如TGF-β1和CTGF的表达,导致ECM的合成与降解失衡,从而抑制肝纤维化。
[Abstract]:Aim: to investigate the effects of caffeine on hepatic fibrosis induced by carbon tetrachloride (CCL 4) in SD rats, and to investigate the possible therapeutic mechanism of caffeine. Methods: 100 SD rats were randomly divided into 4 groups, 25 rats in each group, and subcutaneously injected with olive oil 3ml/ (kg rat body weight) as control group (n = 25), twice a week for 8 weeks. Olive oil containing 5014 was subcutaneously injected into the model group (n = 25) and caffeine treatment group (n ~ (25) twice a week for 8 weeks with 3ml/ (kg body weight), while the control group (n ~ (25) was not treated with liver fibrosis. At the same time, caffeine treatment group and caffeine control group were given caffeine 50 mg / (kg body weight) once a day at the beginning of the first week, and sodium carboxymethyl cellulosein (CMC-Na) was used to treat rats in blank control group and model group. Methods caffeine was used for 8 weeks. At the ninth week, the rats were killed and weighed after anesthesia. The liver tissues were stained with HE and Masson respectively, and the liver tissues were stained with HE and Masson. The expression of alpha-Smooth muscle actin (伪 -SMA) in liver tissue was detected by immunohistochemical staining. Real time quantitative polymerase chain reaction (Real time qPCR) was used to detect the expression of transforming growth factor-beta 1 (TGF- 尾 1), connective tissue growth factor, growth factor (CTGF) and 伪 -SMA. Results: compared with the control group, the expression of TGF- 尾 1 CTGF and 伪 -SMA in the model group was significantly higher than that in the control group (P0.01), and the weight of the rats in the caffeine treatment group was significantly higher than that in the caffeine treatment group (P0.01), and the expression of TGF- 尾 1 CTGF and 伪 -SMA in the model group was significantly higher than that in the control group (P0.01). The expression of TGF- 尾 1 CTGF in hepatic fibrosis was significantly decreased (p0.01), while the expression of 伪 -SMA was not significantly decreased (p0.05), but still statistically significant, compared with that of TGF- 尾 1, the caffeine control group was compared with the blank control group. There was no significant difference in the above data, namely, p0.05. Conclusion: (1) caffeine can inhibit experimental hepatic fibrosis induced by CC14 in SD rats to a certain extent, (2) there is no significant change in liver of healthy rats after continuous caffeine intake for 8 weeks (P0.05). (3) the anti-hepatic fibrosis mechanism of caffeine may be as follows: inhibiting hepatic inflammation, inhibiting the activation of HSCs and MFs (down regulation of a-SMA expression), reducing the expression of hepatic fibrosis promoting factors such as TGF- 尾 1 and CTGF, leading to the imbalance of synthesis and degradation of ECM, thus inhibiting hepatic fibrosis.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.2

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