目的: 研究散结片联合立体定向放射治疗对肝癌大鼠肿瘤组织中PKM2/STAT3通路及巨噬细胞极化的影响。方法: 健康SD雄性大鼠50只,按照随机数字表法分为健康组(健康大鼠正常喂养)、肝癌组(肝癌模型大鼠)、散结片组(模型大鼠+散结片)、放疗组(模型大鼠+定向放疗)、联合组(模型大鼠+散结片+定向放疗),每组10只。采用RH-35细胞肝小叶内注射制备大鼠肝癌模型。HE染色观察肝癌大鼠病理组织形态;TUNEL检测肝癌细胞凋亡;免疫组化检测CD11c、CD206表达水平;免疫印迹检测PKM2 、STAT3与p-STAT3表达水平。结果: 健康组大鼠肝细胞排列有序,形态正常;肝癌组大鼠肝癌细胞生长旺盛,细胞核大小不一,总体上细胞生长分布均匀且紧密,肿瘤组织间隙有大量毛细血管分布。散结片组、放疗组、联合组大鼠肿瘤细胞有不同程度的空泡,肿瘤组织的排列更为疏松,癌细胞有所减少。与健康组大鼠相比,肝癌组大鼠CD11c水平降低,CDK4、CyclinD1、CD206、PKM2、p-STAT3与STAT3水平升高(P<0.05)。与肝癌组大鼠相比,散结片组与放疗组大鼠细胞凋亡、CD11c水平升高,CDK4、CyclinD1、CD206、PKM2、p-STAT3与STAT3水平降低(P<0.05)。与放疗组大鼠相比,联合组大鼠细胞凋亡、CD11c水平升高,CDK4、CyclinD1、CD206、PKM2、p-STAT3、STAT3水平降低(P<0.05)。结论: 散结片联合定向放疗通过抑制PKM2/STAT3通路,促进巨噬细胞向M1型极化并抑制其向M2型极化,促进肝癌细胞的凋亡并抑制其增殖。

Objective: To study the effects of Sanjie tablet combined with stereotactic radiotherapy on the PKM2/STAT3 signaling pathway and the polarization of macrophages in the tumor tissues of hepatoma rats. Methods: Fifty healthy SD male rats were randomly divided into healthy group (normal feeding of healthy rats),hepatocellular carcinoma group (hepatocellular carcinoma rat model),Sanjie tablet group (model+Sanjie tablet),radiotherapy group (model+directional radiotherapy),combined group(model+Sanjie tablet+directional radiotherapy),with 10 rats in each group.He staining was used to observe the pathological morphology of liver cancer rats.Apoptosis of hepatoma cells was detected by TUNEL.CD11c and CD206 levels were detected by immunohistochemistry.The levels of PKM2,STAT3 and p-STAT3 were detected by Western blotting. Results: The liver cells of the healthy group were orderly and normal in shape.Hepatocellular carcinoma cells of rats in the hepatocellular carcinoma group grew vigorously,and their nuclei were of different sizes.On the whole,the cell growth was evenly and closely distributed,and there were a large number of capillaries in the space between tumor tissues.The tumor cells were vacuolated in different degrees,the arrangement of tumor tissue was more loose,and the cancer cells were reduced.Compared with healthy group,apoptosis and CD11c levels were decreased in hepatocellular carcinoma group,while CDK4,CyclinD1,CD206,PKM2,p-STAT3 and STAT3 levels were increased (P<0.05).Compared with liver cancer group,the levels of apoptosis and CD11c were increased,and the levels of CDK4,CyclinD1,CD206,PKM2,p-STAT3 and STAT3 were decreased in the loose knot group and radiotherapy group (P<0.05).Compared with radiotherapy group,apoptosis and CD11c levels were increased,while CDK4,CyclinD1,CD206,PKM2,p-STAT3 and STAT3 levels were decreased in combined group (P<0.05). Conclusion: Sanjie tablet combined with targeted radiotherapy can promote the polarization of macrophages to M1 type and inhibit the polarization of macrophages to M2 type,promote the apoptosis and inhibit the proliferation of hepatoma cells by inhibiting the PKM2/STAT3 pathway.