Department of Vascular Surgery, No. 1 People's Hospital of Jining City, Shandong Province, China.
Eur Rev Med Pharmacol Sci. 2017 Dec;21(24):5813-5820. doi: 10.26355/eurrev_201712_14029.
To investigate the effects of carvedilol on inflammation, apoptosis, and hepatic fibrosis caused by biliary cirrhosis and its mechanisms in mice.
60 male C57/BL6 mice were randomly divided into sham-operation group (Sham group, n=20), biliary cirrhosis group (BDL group, n=20) and carvedilol group (CAR group, n=20). The CAR group was treated with gavage using 12.5 mg/kg carvedilol, once a day for 14 consecutive days, while the Sham group and BDL group were treated with gavage using the equivalent normal saline. After that, the mice in Sham group received the laparotomy under chloral hydrate anesthesia, followed by direct abdominal closure. The mice in BDL group and CAR group received the common bile duct ligation after anesthesia for modeling. After modeling, the survival rate of mice in each group was detected, and the blood and liver tissues were taken for detection. The morphological changes in liver tissues and apoptosis in mice in each group were detected and compared. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), superoxide dismutase (SOD), malondialdehyde (MDA), hydroxyproline, and α-smooth muscle actin (α-SMA) were also detected. The mRNA expression levels of pro-inflammatory factors, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), transforming growth factor β-1 (TGF-β1), α-SMA and collagen-1 were detected via reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of CHOP (CCAAT-enhancer-binding protein homologous protein), activating transcription factor 4 (ATF4), ATF6, inositol-requiring enzyme 1 (IRE1), phosphorylated Jun N-terminal kinase (pJNK), α-SMA, and collagen-1, were detected via Western-blotting.
Our study showed that carvedilol could significantly alleviate the biliary cirrhosis in mice, and improve the survival rate of mice. The ALT, AST and TBIL levels, severity of cirrhosis, and number of apoptotic cells in CAR group were significantly lower than those in BDL group. The levels of α-SMA and hydroxyproline in CAR group were also significantly lower than those in BDL group. The activity of SOD in CAR group was significantly higher than that in BDL group; the above differences were statistically significant (p<0.05). In addition, it was also found that carvedilol could down-regulate the mRNA expression levels of iNOS, COX-2 and TGF-β1, down-regulate the mRNA and protein expression levels of α-SMA and collagen-1, and negatively regulate the ATF4-CHOP, ATF6-CHOP and IRE1-pJNK signaling pathways.
Carvedilol has a significant effect on alleviating the biliary cirrhosis in mice, and its relevant mechanism may be that carvedilol inhibits the endoplasmic reticulum stress through the negative regulation of ATF4-CHOP, ATF6-CHOP and IRE1-pJNK signaling pathways, which needs to be confirmed by further in vitro experiments.
探讨卡维地洛对胆源性肝硬化引起的炎症、凋亡和肝纤维化的影响及其机制。
60 只雄性 C57/BL6 小鼠随机分为假手术组（Sham 组，n=20）、胆源性肝硬化组（BDL 组，n=20）和卡维地洛组（CAR 组，n=20）。CAR 组每天给予 12.5mg/kg 的卡维地洛灌胃，连续 14 天，而 Sham 组和 BDL 组则给予等量的生理盐水灌胃。之后，Sham 组在氯醛麻醉下接受剖腹手术，然后直接关闭腹部。BDL 组和 CAR 组在麻醉后行胆总管结扎术进行建模。建模后，检测各组小鼠的生存率，并采集血液和肝脏组织进行检测。比较各组小鼠肝组织的形态变化和凋亡情况。检测各组小鼠丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、超氧化物歧化酶（SOD）、丙二醛（MDA）、羟脯氨酸和α-平滑肌肌动蛋白（α-SMA）水平。采用逆转录-聚合酶链反应（RT-PCR）检测各组促炎因子诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）、转化生长因子-β1（TGF-β1）、α-SMA 和胶原-1 的 mRNA 表达水平。采用 Western-blotting 检测各组 CHOP（CCAAT 增强子结合蛋白同源蛋白）、激活转录因子 4（ATF4）、ATF6、肌醇需求酶 1（IRE1）、磷酸化 Jun N 末端激酶（pJNK）、α-SMA 和胶原-1 的蛋白表达水平。
本研究表明卡维地洛能显著缓解小鼠的胆源性肝硬化，提高小鼠的生存率。CAR 组的 ALT、AST 和 TBIL 水平、肝硬化严重程度和凋亡细胞数量均明显低于 BDL 组。CAR 组的α-SMA 和羟脯氨酸水平也明显低于 BDL 组。CAR 组的 SOD 活性明显高于 BDL 组；上述差异均有统计学意义（p<0.05）。此外，还发现卡维地洛可下调 iNOS、COX-2 和 TGF-β1 的 mRNA 表达水平，下调α-SMA 和胶原-1 的 mRNA 和蛋白表达水平，并负调控 ATF4-CHOP、ATF6-CHOP 和 IRE1-pJNK 信号通路。
卡维地洛对缓解小鼠胆源性肝硬化有显著作用，其相关机制可能是通过负调控 ATF4-CHOP、ATF6-CHOP 和 IRE1-pJNK 信号通路抑制内质网应激，这需要通过进一步的体外实验来证实。
