槲芪散对肝癌前病变端粒酶活性的调控作用

Regulatory effect of a Chinese medicine prescription Hu Qi Shan on the telomerase activity in hepatic precancerous lesions in rats

目的探讨端粒酶在肝癌前病变形成和发展中的作用及方剂槲芪散(HQS)、君药槲寄生对其活性的调控机制。方法将大鼠分为模型组、HQS大剂量组[8 g/(kg·d)]、小剂量组[4 g/(kg·d)]、槲寄生碱组[8 mg/(kg·d)]及正常组。运用经典的Solt-Farber二步法复制大鼠肝癌前病变的模型,采用组织化学方法检测肝脏组织中γ-谷氨酰转肽酶活性的表达;免疫荧光的方法检测肝脏组织中AFP的表达;采用Quantitative Telomerase Detection Kit(QTD Kit)测定肝脏组织中端粒酶的活性;免疫组织化学方法检测肝脏组织中NF-κB P65蛋白的表达;Western blot的方法测定IκB-α在胞浆蛋白中的含量。结果 HQS和槲寄生总碱治疗后,肝脏中γ-GT阳性灶面积、AFP的阳性细胞数均较模型组明显减少(P<0.05);同时显示治疗组大鼠肝脏中NF-κB P65的阳性细胞数较模型组相比明显减少(P<0.05);治疗后IκB-α在胞浆中的蛋白含量与模型组相比有所增加,差异有显著性(P<0.05)。结论 HQS和槲寄生总碱能够抑制肝癌前病变组织端粒酶活性的表达,其作用是通过抑制凋亡相关基因NF-κB的过表达,增加IκB-α的表达,进而降低端粒酶的活性。

Objective To explore the possible mechanism of action of telomerase in hepatic precancerous lesions,and the regulatory effect of a Chinese medicine prescription HU Qi Shan（ HQS） and its principal drug mistletoe alkali on the telomerase activity in rat liver tissues. Methods Rat model of hepatic precancerous lesions was established by SoltFarber two-step protocol. The model rats were randomly divided into 5 groups,including the model group,high-dose HQS[8 g/（ kg·d） ] group,low-dose HQS [4 g/（ kg·d） ] group,and mistletoe alkali[8 mg/（ kg·d） ] group. γ-Glutamytranspeptidase（ γ-GT） was analyzed by immunohistochemistry. AFP was detected by immunofluorescence technique. The telomerase activity was detected using a quantitative telomerase detection kit. The expression of NF-κB P65 was detected by immunohistochemistry. The cytoplasmic protein IκB-α was detected by western blotting. Results After treated with HQS and mistletoe alkali,the areas of γ-GT-positive foci and number of AFP-positive cells in the liver tissus were significantly decreased than those of the model group（ P〈0. 05 for both）,the telomerase activity was decreased,the number of NF-κB P65-positive cells was also decreased（ P〈0. 05）, whereas the intracytoplasmic expression of IκB-α proteins was significantly increased（ P〈0. 05）. Conclusions HQS and mistletoe alkali can suppress the telomerase activity. Its possible mechanism may be through inhibition of the over-expressed apoptosis-related genes such as NF-κB P65 and increase the expression of IκB-α decreasing the telomerase activity.