Department of Rehabilitation, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, China.
Department of General Surgery (Hepatobiliary Surgery), The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan Province, China.
Sci Rep. 2022 Nov 7;12(1):18873. doi: 10.1038/s41598-022-23539-5.
The study objective was to observe the treatment effect of the farnesoid X receptor (FXR) agonist GW4064 in a rat model of hilar cholangiocarcinoma to explore a new therapeutic target for gene therapy for hilar cholangiocarcinoma. Eighty male Wistar rats were randomly divided into four groups (treatment group, model group, control group and sham operation group, 20 rats in each group). The four groups were fed a standard diet. The treatment group and the model group were injected with a suspension of cholangiocarcinoma QBC939 cells into the hilar bile duct with a microsyringe, the control group was injected with normal saline, and the sham operation group was not injected with anything. A modified tail suspension test (TST) was used to evaluate the vitality of the rats. At 4 weeks, one rat in the treatment group and model group was euthanized, and the changes in the hilar bile duct were recorded. The procedure was repeated at 6 weeks. After 6 weeks, hilar cholangiocarcinoma occurred in the treatment group and model group. Then, the treatment group was injected with GW4064 intraperitoneally at a dose of 50 mg/kg/day. One week after injection, the rats in the four groups were euthanized. Pathological examination confirmed that tumours had formed, and hilar bile duct tissues were taken from the four groups. FXR, Bsep, Ntcp and NF-κB expression in the hilar bile duct was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry. After three weeks, the rats in the treatment group and model group ate less, and their weight was significantly reduced. Six weeks later, hilar cholangiocarcinoma was detected in the treatment group and model group. After treatment with GW4064, the ratios of FXR/GAPDH mRNA, Bsep/GAPDH mRNA, Ntcp/GAPDH mRNA and NF-κBp65/GAPDH mRNA were significantly different among the four groups. Under a light microscope, FXR protein reacted with anti-FXR antibody, Bsep protein reacted with anti-Bsep antibody, Ntcp protein reacted with anti-Ntcp antibody and NF-κBp65 protein reacted with anti-NF-κBp65 antibody, and they showed granular expression. Every pathological section included 4,800 cells, and there were different numbers of positive cells in each group. FXR expression in the hilar cholangiocarcinoma of rats was significantly lower than that in normal hilar bile duct tissues. GW4064 increased the expression of FXR in tumour tissues. These findings suggest that FXR may be a new therapeutic target and that GW4064 may be helpful in the treatment of hilar cholangiocarcinoma.
研究目的在于观察法尼醇 X 受体（FXR）激动剂 GW4064 在大鼠肝门部胆管癌模型中的治疗效果，探索肝门部胆管癌基因治疗的新治疗靶点。将 80 只雄性 Wistar 大鼠随机分为 4 组（治疗组、模型组、对照组和假手术组，每组 20 只）。4 组大鼠均给予标准饮食。治疗组和模型组采用微量注射器将胆管癌细胞 QBC939 混悬液注入肝门胆管，对照组注入生理盐水，假手术组不注射任何药物。采用改良的悬尾试验（TST）评估大鼠活力。4 周时，治疗组和模型组各处死 1 只大鼠，记录肝门胆管变化。6 周时重复该过程。6 周后，治疗组和模型组大鼠发生肝门部胆管癌。然后，治疗组大鼠腹腔注射 GW4064 50mg/kg/d。注射 1 周后，4 组大鼠均处死。病理检查证实肿瘤形成，从 4 组大鼠取肝门胆管组织。采用实时聚合酶链反应（RT-PCR）和免疫组织化学法检测肝门胆管中 FXR、Bsep、Ntcp 和 NF-κB 的表达。3 周后，治疗组和模型组大鼠摄食量减少，体重明显减轻。6 周后，治疗组和模型组大鼠发生肝门部胆管癌。GW4064 治疗后，4 组大鼠 FXR/GAPDHmRNA、Bsep/GAPDHmRNA、Ntcp/GAPDHmRNA 和 NF-κBp65/GAPDHmRNA 的比值差异有统计学意义。光镜下，FXR 蛋白与抗 FXR 抗体反应，Bsep 蛋白与抗 Bsep 抗体反应，Ntcp 蛋白与抗 Ntcp 抗体反应，NF-κBp65 蛋白与抗 NF-κBp65 抗体反应，均呈颗粒状表达。每个病理切片包含 4800 个细胞，每组有不同数量的阳性细胞。大鼠肝门部胆管癌组织中 FXR 表达明显低于正常肝门胆管组织。GW4064 增加了肿瘤组织中 FXR 的表达。这些发现提示 FXR 可能成为新的治疗靶点，GW4064 可能有助于肝门部胆管癌的治疗。
