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目的观察粒-巨噬细胞集落刺激因子(granulocyte macrophage-colony stimulating factor,GM-CSF)局部应用于草酸铂腹腔化疗下大鼠结肠吻合口愈合过程中的组织学变化,探讨其作用机制。方法 10周龄清洁级雄性Wistar大鼠60只,体重(235±18)g,制备结肠吻合模型,随机分为3组(n=20)。术后1 d,A、B组腹腔对应注射10 mL 5%葡萄糖溶液、10 mL含草酸铂(25 mg/kg)的5%葡萄糖溶液;C组术后即刻吻合口区域注射50μg GM-CSF,术后1 d腹腔注射10 mL含草酸铂(25 mg/kg)的5%葡萄糖溶液。术后观察大鼠一般情况,于2、3、5、7 d测量吻合口破裂压,组织学观察并计算吻合口愈合指数,免疫组织化学染色检测Ⅰ型胶原纤维阳性染色面积。结果术后各组大鼠均存活至实验完成。术后2、3 d各组吻合口破裂压相似(P>0.05);5、7 d时,B组吻合口破裂压显著低于A、C组(P<0.05),A、C组间差异无统计学意义(P>0.05)。吻合口愈合指数结果示,术后各时间点,A、C组间单核细胞浸润、黏膜上皮细胞覆盖、黏膜下层-肌层衔接程度及肉芽组织形成相似(P>0.05),A、C组黏膜上皮细胞覆盖及肉芽组织形成均显著优于B组(P<0.05);术后2、3 d,A、C组单核细胞浸润显著优于B组(P<0.05),5、7 d黏膜下层-肌层衔接程度优于B组(P<0.05)。术后2、3 d,各组Ⅰ型胶原纤维阳性染色面积比较差异均无统计学意义(P>0.05);5、7 d时A、C组Ⅰ型胶原纤维阳性染色面积显著高于B组(P<0.05),A、C组间比较差异无统计学意义(P>0.05)。结论局部注射GM-CSF可通过早期动员巨噬细胞浸润,提高大鼠黏膜下层胶原纤维沉积,从而加速结肠愈合进程。
Objective To observe the histological changes of granulocyte macrophage-colony stimulating factor (GM-CSF) in the healing process of colonic anastomosis in rats undergoing oxaliplatin intraperitoneal chemotherapy and to explore its mechanism. Methods Sixty male Wistar rats aged 10 weeks were randomly divided into 3 groups (n = 20). The body weight was (235 ± 18) g. At 1 d after operation, 10 mL of 5% dextrose solution and 10 mL of 5% dextrose solution containing oxaliplatin (25 mg / kg) were intraperitoneally injected into the abdominal cavity of group A and group B. Group C received 50 μg of GM-CSF immediately after the operation, One ml of 5% dextrose solution containing oxaliplatin (25 mg / kg) was injected intraperitoneally 1 d after operation. Postoperatively, the general condition of the rats was observed. The ankle rupture pressure was measured at 2, 3, 5 and 7 days. Histological observation and healing index were calculated. The positive staining area of collagen type Ⅰ was detected by immunohistochemical staining. Results After operation, all rats survived until the experiment was completed. The anastomotic rupture pressure in each group was similar at 2 and 3 days after operation (P> 0.05). At 5 and 7 days, the ankle rupture pressure in group B was significantly lower than that in group A and C (P <0.05) No statistical significance (P> 0.05). The results of anastomotic healing index showed that mononuclear cell infiltration, mucosal epithelial cell coverage, submucosal-myometrial cohesion and granulation tissue formation were similar in groups A and C (P> 0.05) Mucosal epithelial cells and formation of granulation tissue were significantly better than those in group B (P <0.05). Monocytes infiltrated in group A and C at 2 and 3 days after operation were significantly better than those in group B (P <0.05) Submucosa - myometrial convergence better than the B group (P <0.05). There was no significant difference in the positive staining area of type Ⅰ collagen fibers between the two groups at postoperative day 2 and 3 (P> 0.05). On the 5th and 7th day, the positive staining area of collagen Ⅰ in group A and C was significantly higher than that of group B (P <0.05). There was no significant difference between A and C groups (P> 0.05). Conclusion Local injection of GM-CSF accelerates colonic healing through early mobilization of macrophage infiltration, increasing collagen deposition in the submucosa of rats.