Wan Xuedong, Fang Yuanyuan, Qin Minjing, Zheng Qitong, Yang Qiao, Peng Mengyun, Hao Min, Wang Kuilong, Zhao Ruihua, Shi Yiqing, Han Xin, Sang Xia'nan, Cao Gang
School of Pharmacy, Zhejiang Chinese Medical University, Hangzhou, China.
Songyang Institute, Zhejiang Chinese Medical University, Lishui, China.
Front Pharmacol. 2024 Sep 20;15:1479503. doi: 10.3389/fphar.2024.1479503. eCollection 2024.
Hepatic fibrosis and its associated consequences continue to pose a substantial global health challenge. Developing novel approaches to hepatic fibrosis management and prevention is critically necessary. Radix Paeoniae Alba (RPA) is widely used in Traditional Chinese Medicine (TCM) to treat various diseases. Our earlier research found that a bioactive component of RPA had a dose-dependent effect on anti-allergic asthma. RPA reduces allergic asthma by slowing the hepatic wind, according to "Treatise on Febrile Diseases". However, this bioactive fraction's pharmacological effects and mechanisms on the liver are unknown.
This study examined the bioactive fraction MP-40, the methanol extract of RPA (MRPA), on bile duct ligation (BDL) for its anti-hepatic fibrosis activity and potential mechanisms.
First, the effectiveness of MP-40 in treating BDL-induced hepatic fibrosis in mice and rats was evaluated through survival rates, ALT, AST HYP, and pathological changes. Molecular assays were performed using cultures of HSC-T6 activation. The expression of α-SMA and Collagen I evaluated fibro-tropic factors with HSC activation. Furthermore, the levels of pyroptosis were assessed by examining the expression of the pyroptosis-related proteins, including NLRP3, Cleaved Caspase-1, GSDMD-N, and 1L-1β. Additionally, the effective constituents of MP-40 were identified by extraction, separation, and identification. Finally, PF and TGG, as the delegate compounds of MP-40, were tested to confirm their inhibition effects on HSC-T6 activation.
The findings demonstrated that MP-40 and MRPA could lower ALT, AST, and HYP levels, boost survival rates, and reduce liver damage in BDL mice and rats. Furthermore, MP-40 outperforms MRPA. MP-40 was proven to drastically diminish fibrotic α-SMA and Collagen I. The expression of pyroptosis-related proteins NLRP3, Cleaved Caspase-1, TGF-β1, GSDMD-N, and 1L-1β decreased. MP-40 inhibited the synthesis of pyroptosis-related proteins more effectively than MCC950 (an NLRP3-specific inhibitor). Monoterpene glycosides and tannins were shown to be the most potent MP-40 components. Finally, the delegate compounds MP-40, PF, and TGG were shown to have substantial inhibitory effects on HSC-T6 activation.
The results proved that MP-40 alleviates BDL-induced cholestatic hepatic fibrosis by inhibiting NLRP3-mediated pyroptosis. PF and TGG play a role in treating BDL-induced cholestatic hepatic fibrosis in MP-40.
肝纤维化及其相关后果仍然是全球重大的健康挑战。开发新的肝纤维化管理和预防方法至关重要。白芍在传统中医中广泛用于治疗各种疾病。我们早期的研究发现，白芍的一种生物活性成分对过敏性哮喘有剂量依赖性作用。根据《伤寒论》，白芍通过缓肝风来减轻过敏性哮喘。然而，这种生物活性成分对肝脏的药理作用和机制尚不清楚。
本研究考察了白芍甲醇提取物中的生物活性成分MP-40对胆管结扎（BDL）所致肝纤维化的抗肝纤维化活性及其潜在机制。
首先，通过生存率、谷丙转氨酶（ALT）、谷草转氨酶（AST）、羟脯氨酸（HYP）及病理变化评估MP-40对小鼠和大鼠BDL诱导的肝纤维化的治疗效果。利用肝星状细胞（HSC）-T6激活培养物进行分子检测。通过评估α-平滑肌肌动蛋白（α-SMA）和I型胶原蛋白的表达来检测HSC激活的促纤维化因子。此外，通过检测包括NLRP3、裂解的半胱天冬酶-1、Gasdermin D-N（GSDMD-N）和白细胞介素-1β（IL-1β）在内的焦亡相关蛋白的表达来评估焦亡水平。此外，通过提取、分离和鉴定确定MP-40的有效成分。最后，以PF和TGG作为MP-40的代表化合物，测试它们对HSC-T6激活的抑制作用。
研究结果表明，MP-40和白芍甲醇提取物（MRPA）可降低BDL小鼠和大鼠的ALT、AST和HYP水平，提高生存率，并减轻肝损伤。此外，MP-40的效果优于MRPA。事实证明，MP-40能显著减少纤维化的α-SMA和I型胶原蛋白。焦亡相关蛋白NLRP3、裂解的半胱天冬酶-1、转化生长因子-β1（TGF-β1）、GSDMD-N和IL-1β的表达降低。MP-40比MCC950（一种NLRP3特异性抑制剂）更有效地抑制焦亡相关蛋白的合成。单萜糖苷和单宁被证明是MP-４０中最有效的成分。最后，代表化合物MP-40、PF和TGG对HSC-T6激活具有显著的抑制作用。
结果证明，MP-40通过抑制NLRP3介导的焦亡减轻BDL诱导的胆汁淤积性肝纤维化。PF和TGG在MP-40治疗BDL诱导的胆汁淤积性肝纤维化中发挥作用。
