Carfilzomib改善肿瘤恶病质肝脏功能及其分子机制

Carfilzomib ameliorates hepatic function in mice with cancer cachexia and its molecular mechanism

目的探讨蛋白酶体抑制剂(carfilzomib,CFZ)改善肿瘤恶病质肝脏功能及其分子机制。方法 40只BALB/c小鼠按随机数字表法分为4组,分别是健康组(HC)、CFZ预防组(CP)、CFZ治疗组(CT)、恶病质组(CC),每组10只。荷瘤组通过皮下接种小鼠结肠癌C26细胞构建恶病质模型。接种后第5、12天CP组、CT组分别开始腹腔注射CFZ,第19天处死取标本。自动生化分析仪检测肝功能,ELISA检测肝脏炎症细胞因子和CRP(C-reactive protein),qRT-PCR(quantitative real time polymerase chain reaction)和WB(western blot)检测肝脏IκBα和p65 mRNA和蛋白质。结果 CC组小鼠白蛋白较HC组、CP组、CT组显著降低,CC组谷丙转氨酶、谷草转氨酶、总胆红素、直接胆红素、甘油三酯浓度分别为(221.67±12.38)U/L、(315.53±13.60)U/L、(1.65±0.32)mmol/L、(0.88±0.21)mmol/L、(4.98±0.32)mmol/L;经CFZ处理后,CP组、CT组谷丙转氨酶、谷草转氨酶、总胆红素、直接胆红素、甘油三酯浓度分别为(108.27±16.55)U/L、(180.45±15.28)U/L、(1.15±0.27)mmol/L、(0.58±0.12)mmol/L、(2.93±0.18)mml/L和(148.56±18.16)U/L、(247.18±21.64)U/L、(1.34±0.19)mmol/L、(0.69±0.16)mml/L、(3.75±0.28)mmol/L,CP组改善作用较CT组更明显。CC组中TNFa、IL-1、IL-6、CRP浓度为(156±9.56)ng/L、(762±9.46)ng/L、(962±9.12)ng/L、(772±10.04)ng/L,与HC组[(16.42+5.63)ng/L、(174+9.61)ng/L、(206±8.27)ng/L、(397±10.2)ng/L)]、CP组[(71.25±4.41)ng/L、(398±9.72)ng/L、(398±9.72)ng/L、(483±9.71)ng/L)]、CT组[(113±8.01)ng/L、(506±8.74)ng/L、(703±7.76)ng/L、(651±11.31)ng/L)]比较明显升高。与CC组比较,HC组、CP组和CT组IκBα表达量明显升高,差异有统计学意义,CP组升高比CT组更明显(P=0.000);与HC组比较,CP组、CT组和CC组p65表达量明显升高,差异有统计学意义,CT组较CP组升高明显(P=0.000)。结论 CFZ改善肿瘤恶病质小鼠肝脏功能可能与抑制NF-κB而降低肝脏炎症,抑制肿瘤生长和骨骼肌消耗有关。

Objective To study the role of CFZ in the amelioration of hepatic function in cancer cachexia and its associated mechanism. Methods Forty BALB/c mice were selected. BALB/c mice were divided into 4 groups randomly, including a healthy control group (HC),a CFZ prevention group (CP), a CFZ treatment group (CT) and a cancer cachexia group (CC).Cancer cachexia model was induced by injecting murine colon 26 adencarcinoma cells into male BALB/c mice intraperitoneally. Following administration of CFZ intraperitoneally twice a week to CP and CT groups on the days 5 and 12 after tumor cells injection, respectively,all mice were acrificed on day 19. hepatic function was detected by automatic biochemistry analyzer, The concentration of inflammatory cytokines and CRP were detected by ELISA. The mRNA and protein expression of IκBα and p65 were detected by real-time PCR and western blotting. Results Compared with HC group, CP group and CT group, the albumin in CC group was significantly decreased, and the concentration of glutamate transaminase, total bilirubin, direct bilirubin and triglyceride were significantly increased,(221.67±12.38) U/L、(315.53±13.60) U/L、(1.65±0.32) mmol/L、(0.88±0.21) mmol/L、(4.98±0.32) mmol/L respectively. Liver biochemical test of CP group[(108.27±16.55) U/L、(180.45±15.28) U/L、(1.15±0.27) mmol/L、(0.58±0.12) mmol/L、(2.93±0.18) mml/L) and CT group [(148.56±18.16) U/L、(247.18±21.64) U/L、(1.34±0.19) mmol/L、(0.69±0.16) mml/L、(3.75±0.28) mmol/L] was improved after CFZ treatment, and CP group was better than that of CT group. The concentrations of TNFa, IL-1, IL-6 and CRP in CC group[(156±9.56) ng/L、(762±9.46) ng/L、(962±9.12) ng/L、(772±10.04 ng/L)] were significantly higher than those in HC group[(16.42±5.63ng/L、174±9.61 ng/L、206±8.27 ng/L、397±10.2 ng/L)], CP group[(71.25±4.41 ng/L、398±9.72 ng/L、398±9.72 ng/L、483±9.71 ng/L)] and CT group[(113±8.01 ng/L、506±8.74 ng/L、703±7.76ng/L、651±11.31 ng/L)]. The expression of IκBα in HC group, CP group and CT group were higher than that in CC group, and the difference was statistically significant. The expression of IκBαwas was more obvious in CP group than that in CT group. Compared with HC group, the expression of p65 in CP group, CT group and CC group was significantly increased, respectively, and the difference was statistically significant. The expression of p65 in CP group was lower than that in CT group (P=0.000). Conclusion CFZ ameliorates hepatic function in cancer cachexia mice, which may be related to the inhibition of NF-κB resulting in liver function improvement, the inhibition of tumor growth and the consumption of skeletal muscle.