Introduction: Liver disease is common and often life-threatening. Sinomenine (SIN) is an active ingredient extracted from Sinomenium acutum. This study investigated the protective effect and mechanism of sinomenine (SIN) on acetaminophen (APAP)-induced liver injury from in vitro and in vivo.
Methods: In vivo experiments, mice were randomly divided into six groups (n=10): control group, model group, SIN (25 mg/kg) group, SIN (50 mg/kg) group, SIN (100 mg/kg) group and SIN (100 mg/kg) + SRI-011381 group. Alanine transaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP) were detected. The pathological lesion was measured by HE staining. Apoptosis was measured by TUNEL staining. In vitro experiments, BRL-3A cells were treated with APAP (7.5 mM) and then subjected to various doses of SIN (10, 50 and 100 μg/mL) at 37°C for 24 h. Inflammatory factors and oxidative stress index were measured by ELISA. The expression of proteins was detected by Western blot.
Results: The results showed that compared with the control group, the levels of ALT, AST and ALP in the serum of APAP-induced mice were significantly increased, followed by liver histological damage and hepatocyte apoptosis. Besides, APAP reduced the activity of SOD and GSH-Px, while increasing the content of MDA and LDH. Notably, APAP also promoted the expression of NLRP3, ASC, caspase-1 and IL-1β. Interestingly, SIN treatment dose-dependently reduced APAP-induced liver injury and oxidative stress, inhibited the activation of NLRP3 inflammasomes, and reduced the levels of inflammatory cytokines. In vitro studies have shown that SIN treatment significantly reduced the viability of BRL-3A cells and oxidative stress and inflammation. In addition, the Western blotting analysis showed that SIN inhibited the activation of TGF-β/Smad pathway in a dose-dependent manner in vitro and in vivo. These effects were significantly reversed by TGF-β/Smad activator SRI-011381 or TGF-β overexpression.
Discussion: The study indicates that SIN attenuates APAP-induced acute liver injury by decreasing oxidative stress and inflammatory response via TGF-β/Smad pathway in vitro and in vivo.

Keywords: Sinomenine, acetaminophen, inflammatory response, oxidative stress, TGF-β/Smad pathway, acute liver injury


中文翻译：

---

Sinomenine 通过在体外和体内调节 TGF-β/Smad 通路降低氧化应激和炎症反应，减轻对乙酰氨基酚引起的急性肝损伤。

简介：肝病很常见，而且常常危及生命。青藤碱（SIN）是从青藤中提取的活性成分。本研究从体内外研究了青藤碱（SIN）对对乙酰氨基酚（APAP）诱导的肝损伤的保护作用和机制。
方法：在体内实验中，将小鼠随机分为六组（n=10）：对照组、模型组、SIN（25 mg/kg）组、SIN（50 mg/kg）组、SIN（100 mg/kg）组和SIN (100 mg/kg) + SRI-011381 组。检测到丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和碱性磷酸酶（ALP）。HE染色检测病理病变。通过 TUNEL 染色测量细胞凋亡。在体外实验中，BRL-3A 细胞用 APAP (7.5 mM) 处理，然后在 37°C 下经受各种剂量的 SIN (10、50 和 100 μg/mL) 24 小时。ELISA法测定炎症因子和氧化应激指数。Western blot检测蛋白表达。
结果：结果显示，与对照组相比，APAP诱导小鼠血清中ALT、AST、ALP水平明显升高，其次是肝组织学损伤和肝细胞凋亡。此外，APAP降低了SOD和GSH-Px的活性，同时增加了MDA和LDH的含量。值得注意的是，APAP 还促进了 NLRP3、ASC、caspase-1 和 IL-1β 的表达。有趣的是，SIN 治疗剂量依赖性地减少了 APAP 诱导的肝损伤和氧化应激，抑制了 NLRP3 炎性体的激活，并降低了炎性细胞因子的水平。体外研究表明，SIN 治疗显着降低了 BRL-3A 细胞的活力以及氧化应激和炎症。此外，Western印迹分析显示，SIN在体外和体内均以剂量依赖性方式抑制TGF-β/Smad通路的激活。TGF-β/Smad 激活剂 SRI-011381 或 TGF-β 过表达显着逆转了这些作用。
讨论：该研究表明，SIN 在体外和体内通过 TGF-β/Smad 通路降低氧化应激和炎症反应，从而减轻 APAP 诱导的急性肝损伤。

关键词：青藤碱，对乙酰氨基酚，炎症反应，氧化应激，TGF-β/Smad通路，急性肝损伤