Institute of Nuclear Medicine and Allied Sciences, Defence Research and Development Organisation, Delhi, India.
All India Institute of Medical Sciences, Ansari Nagar, Delhi, India.
Biomed J. 2021 Oct;44(5):611-619. doi: 10.1016/j.bj.2020.05.012. Epub 2020 May 23.
Methotrexate (MTX) is widely used in chemotherapy but its associated hepatotoxicity is a major complication, limiting its use. This study evaluates possible therapeutic effect of oral alpha-ketoglutarate (AKG) supplementation against MTX-induced hepatotoxicity.
HepG2 cells were used to evaluate in-vitro cyto-protection conferred by AKG against MTX induced cytotoxicity. For in-vivo animal study, rats were divided into three groups. Group-I served as control. Group-II animals were administered single intraperitoneal injection of MTX (20 mg/kg/body weight), while Group-III received MTX as in group-II followed by oral AKG (2 gm/kg body weight) for 5 days. Tc-Mebrofenin hepatobiliary study was performed under a gamma camera to determine real time functional status of rats' livers. Multiple parameters concerning hepatic mebrofenin uptake and excretion, including T and T in control and treated animals were determined. Biochemical analysis of the liver homogenate in terms of hepatic enzyme activities in serum, antioxidant status, tissue factor activity, tissue collagen content and histological analysis of the liver tissue were also done.
AKG supplementation significantly reversed MTX induced derangement in activities of serum liver enzymes [ALT and ALP (p = 0.003); AST (p = 0.005)], antioxidant status [LPO and GSH (p = 0.005); CAT (p = 0.004)], tissue factor activity (p = 0.005) and tissue collagen content (p = 0.005). Functional imaging confirmed that hepatic retention and fractional biliary excretion were significantly abnormal in MTX treated group (T: 234 s ± 40 s; T: 846sec ± 32sec) as compared to AKG supplemented group (T: 144 s ± 35sec; T: 468sec ± 27sec). Hepatic extraction fraction (HEF) was 92.2 ± 1.8%, 48.7 ± 2.6% and 69.8 ± 4.3% in control, MTX and AKG supplemented rats respectively.
Tc-mebrofenin imaging strongly suggests therapeutic action of AKG in protecting liver damage by MTX in rats. Functional imaging parameters correlated well with biochemical and histopathological findings.
甲氨蝶呤（MTX）广泛用于化疗，但它的肝毒性是一个主要的并发症，限制了它的使用。本研究评估了口服α-酮戊二酸（AKG）补充剂对 MTX 诱导的肝毒性的可能治疗效果。
使用 HepG2 细胞评估 AKG 对 MTX 诱导的细胞毒性的体外细胞保护作用。对于体内动物研究，将大鼠分为三组。第 I 组作为对照。第 II 组动物单次腹腔注射 MTX（20mg/kg/体重），第 III 组动物给予 MTX 后，每天口服 AKG（2g/kg 体重），连续 5 天。用伽马相机进行 Tc-美贝福芬肝胆显像，以确定大鼠肝脏的实时功能状态。测定了 T 和 T 在对照组和治疗组动物中的肝摄取和排泄的多个参数。还对肝匀浆进行了血清肝酶活性、抗氧化状态、组织因子活性、组织胶原含量的生化分析和肝组织的组织学分析。
AKG 补充剂显著逆转了 MTX 诱导的血清肝酶活性[ALT 和 ALP（p=0.003）；AST（p=0.005）]、抗氧化状态[LPO 和 GSH（p=0.005）；CAT（p=0.004）]、组织因子活性（p=0.005）和组织胶原含量（p=0.005）的紊乱。功能成像证实，MTX 治疗组的肝保留和胆汁排泄分数明显异常（T：234s±40s；T：846sec±32sec），而 AKG 补充组（T：144s±35s；T：468sec±27sec）。肝提取分数（HEF）在对照组、MTX 组和 AKG 补充组大鼠中分别为 92.2±1.8%、48.7±2.6%和 69.8±4.3%。
Tc-美贝福芬成像强烈提示 AKG 对 MTX 诱导的大鼠肝损伤具有治疗作用。功能成像参数与生化和组织病理学发现密切相关。
