Krithika Rajesh, Verma Ramtej J, Shrivastav Pranav S, Suguna Lonchin
Department of Biochemistry, Central Leather Research Institute, Council of Scientific and Industrial Research (CSIR), Chennai, India.
Department of Biochemistry, Central Leather Research Institute, Council of Scientific and Industrial Research (CSIR), Chennai, India ; Zoology Department, Gujarat University, Ahmedabad, India.
J Clin Exp Hepatol. 2011 Sep;1(2):57-67. doi: 10.1016/S0973-6883(11)60123-0. Epub 2011 Nov 9.
Phyllanthus amarus, a traditional herbal liver-protecting medicine, is known to contain an active ingredient phyllanthin. Many research studies and clinical trials performed in the past using this plant have given contentious results which clearly accentuates the need for the standardization of the extracts.
In this study, P. amarus extract was standardized for phyllanthin content by high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) analysis. The preventive role of a standardized extract of P. amarus against CC14-induced hepatotoxicity in vivo and in vitro using mice model and human hepatoma HepG2 cell line, respectively, was investigated.
Phyllanthin was used as a marker phytochemical for the standardization of P. amarus extract. The extracts were verified for phyllanthin content by HPTLC and HPLC. Female mice were orally administered with CCl4 either with or without standardized P. amarus extract in three different doses. Similarly, the cytoprotective role of the standardized extract in vitro was studied in HepG2 cell line.
Oral administration of CCl4 resulted in increased oxidative stress, decreased antioxidative defense, and liver injury. Treatment with P. amarus along with CCl4 significantly mitigated the increase in activities of liver marker enzymes, lipid peroxidation, and bilirubin content. It also increased the antioxidant enzymatic and non-enzymatic defense parameter levels. The results of the in vitro study conducted in HepG2 cells indicated that the hepatotoxin lowered 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (Mil) metabolism and increased the release of transaminases which were corrected with co-incubation with P. amarus.
The study established a significant liver-protecting role of standardized P. amarus extract due to the presence of active ingredient phyllanthin.
苦味叶下珠是一种传统的保肝草药，已知其含有活性成分叶下珠素。过去使用这种植物进行的许多研究和临床试验都得出了有争议的结果，这清楚地凸显了提取物标准化的必要性。
在本研究中，通过高效薄层色谱法（HPTLC）和高效液相色谱法（HPLC）分析，对苦味叶下珠提取物的叶下珠素含量进行标准化。分别使用小鼠模型和人肝癌HepG2细胞系，研究了标准化苦味叶下珠提取物对CC14诱导的体内外肝毒性的预防作用。
叶下珠素用作苦味叶下珠提取物标准化的标志性植物化学物质。通过HPTLC和HPLC验证提取物中叶下珠素的含量。雌性小鼠口服给予CCl4，同时给予或不给予三种不同剂量的标准化苦味叶下珠提取物。同样，在HepG2细胞系中研究了标准化提取物的体外细胞保护作用。
口服CCl4导致氧化应激增加、抗氧化防御能力下降和肝损伤。与CCl4一起使用苦味叶下珠进行治疗可显著减轻肝标志物酶活性、脂质过氧化和胆红素含量的增加。它还提高了抗氧化酶和非酶防御参数水平。在HepG2细胞中进行的体外研究结果表明，肝毒素降低了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）代谢，并增加了转氨酶的释放，而与苦味叶下珠共同孵育可纠正这些变化。
该研究证实了由于活性成分叶下珠素的存在，标准化苦味叶下珠提取物具有显著的保肝作用。
