Wang Ying-Chun, Kong Wei-Zong, Jin Qing-Mei, Chen Juan, Dong Lei
Ying-Chun Wang, Lei Dong, Department of Gastroenterology, the Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710004, Shaanxi Province, China.
World J Gastroenterol. 2015 Sep 21;21(35):10104-12. doi: 10.3748/wjg.v21.i35.10104.
To investigate the effects of salvianolic acid B (Sal B) on the morphological characteristics and functions of liver mitochondria of rats with nonalcoholic steatohepatitis (NASH).
A total of 60 male Sprague-Dawley rats were randomly divided into three groups: (1) a normal group fed a normal diet; (2) an NASH model group; and (3) a Sal B-treated group fed a high-fat diet. Two rats from each group were executed at the end of the 12th week to detect pathological changes. The rats in the Sal B-treated group were gavaged with 20 mL/kg Sal B (1 mg/mL) daily. The model group received an equal volume of distilled water as a control. At the end of the 24th weekend, the remaining rats were executed. Serum biochemical parameters and liver histological characteristics were observed. Malondialdehyde (MDA) and superoxide dismutase (SOD) in the liver were determined. Protein expression of CytC and caspase-3 was determined by immunohistochemistry. The mRNA transcripts of mitofusin-2 (Mfn2) and NF-κB in the liver tissue were detected by real-time PCR. Mitochondrial membrane potential was detected using a fluorescence spectrophotometer. Mitochondrial respiratory function was detected using a Clark oxygen electrode.
The model group showed significantly higher ALT, AST, TG, TC and MDA but significantly lower SOD than the normal group. In the model group, the histological characteristics of inflammation and steatosis were also evident; mitochondrial swelling and crest were shortened or even disappeared. CytC (18.46 ± 1.21 vs 60.01 ± 3.43, P < 0.01) and caspase-3 protein expression (30.26 ± 2.56 vs 83.31 ± 5.12, P < 0.01) increased significantly. The mRNA expression of NF-κB increased (0.81 ± 0.02 vs 0.91 ± 0.03, P < 0.05), whereas the mRNA expression of Mfn2 decreased (1.65 ± 0.31 vs 0.83 ± 0.16, P < 0.05). Mitochondrial membrane potential also decreased and breathing of rats was weakened. Steatosis and inflammation degrees in the treatment group were significantly alleviated compared with those of the model group. In the treatment group, mitochondrial swelling was alleviated. CytC (60.01 ± 3.43 vs 30.52 ± 2.01, P < 0.01) and caspase-3 protein expression (83.31 ± 5.12 vs 40.15 ± 3.26, P < 0.01) significantly decreased. The mRNA expression of NF-κB also decreased (0.91 ± 0.03 vs 0.74 ± 0.02, P < 0.01), whereas the mRNA expression of Mfn2 increased (0.83 ± 0.16 vs 1.35 ± 0.23, P < 0.01). Mitochondrial membrane potential increased and respiratory function was enhanced.
Sal B can treat NASH by protecting the morphological characteristics and functions of liver mitochondria, regulating lipid metabolism, controlling oxidative stress and lipid peroxidation and inhibiting apoptosis.
探讨丹酚酸B（Sal B）对非酒精性脂肪性肝炎（NASH）大鼠肝脏线粒体形态特征及功能的影响。
将60只雄性Sprague-Dawley大鼠随机分为三组：（1）正常饮食的正常组；（2）NASH模型组；（3）高脂饮食的Sal B治疗组。每组在第12周结束时处死2只大鼠以检测病理变化。Sal B治疗组大鼠每日灌胃20 mL/kg Sal B（1 mg/mL）。模型组给予等体积蒸馏水作为对照。在第24周末，处死剩余大鼠。观察血清生化参数和肝脏组织学特征。测定肝脏中丙二醛（MDA）和超氧化物歧化酶（SOD）。通过免疫组化测定细胞色素C（CytC）和半胱天冬酶-3（caspase-3）的蛋白表达。通过实时PCR检测肝脏组织中线粒体融合蛋白2（Mfn2）和核因子κB（NF-κB）的mRNA转录本。使用荧光分光光度计检测线粒体膜电位。使用Clark氧电极检测线粒体呼吸功能。
模型组的谷丙转氨酶（ALT）、谷草转氨酶（AST）、甘油三酯（TG）、总胆固醇（TC）和MDA显著高于正常组，但SOD显著低于正常组。在模型组中，炎症和脂肪变性的组织学特征也很明显；线粒体肿胀，嵴缩短甚至消失。CytC（18.46±1.21对60.01±3.43，P<0.01）和caspase-3蛋白表达（30.26±2.56对83.31±5.12，P<0.01）显著增加。NF-κB的mRNA表达增加（0.81±0.02对0.91±0.03，P<0.05），而Mfn2的mRNA表达降低（1.65±0.31对0.83±0.16，P<0.05）。线粒体膜电位也降低，大鼠呼吸减弱。与模型组相比，治疗组的脂肪变性和炎症程度显著减轻。在治疗组中，线粒体肿胀减轻。CytC（60.01±3.43对30.52±2.01，P<0.01）和caspase-3蛋白表达（83.31±5.12对40.15±3.26，P<0.01）显著降低。NF-κB的mRNA表达也降低（0.91±0.03对0.74±0.02，P<0.01），而Mfn2的mRNA表达增加（0.83±0.16对1.35±0.23，P<0.01）。线粒体膜电位增加，呼吸功能增强。
Sal B可通过保护肝脏线粒体的形态特征及功能、调节脂质代谢、控制氧化应激和脂质过氧化以及抑制细胞凋亡来治疗NASH。
