活血通腑方优化方防治大鼠术后腹腔粘连的最佳有效剂量

Preventive and Therapeutic Effect of Optimized Huoxue Tongfu Formula on Postoperative Peritoneal Adhesion in Rats

摘要:

目的：观察活血通腑方优化方对腹腔粘连的治疗效果，确定活血通腑方优选方的最佳有效剂量。方法：取48只雄性SD大鼠随机分为假手术组、模型组、活血通腑方优化方2，6，18 g·kg^-1组、阳性对照四磨汤（5.4 g·kg^-1）组，除假手术组外，其余动物均采用锉刀法制备腹腔粘连大鼠模型，于术后第1天各组分别予以相应剂量的药物灌胃，假手术组给予等量的生理盐水，连续给药1周。术后7 d通过观察大鼠一般情况及腹腔粘连评分，酶联吸附法（ELISA）测定血清白介素-8（IL-8）、白介素-10（IL-10）和肿瘤坏死因子-α（TNF-α）含量，同时实时荧光定量（RT-PCR）检测粘连组织中Ⅰ型胶原（Col-Ⅰ）mRNA的表达。结果：与假手术组相比，模型组肠壁各层血管高度扩张、水肿，浆膜层明显有淋巴细胞浸润、纤维母细胞增生；与模型组相比，各治疗组粘连处肠壁组织形态学上肌层结构完好，浆膜层轻度增厚，浆膜面纤维结缔组织增生不明显。与模型组相比，活血通腑方优化方中剂量组血清IL-8和TNF-α水平分别为（154.2±16.2），（58.2±6.4）ng·L^-1，但是IL-10水平（58.2±6.4）ng·L^-1升高，同时降低粘连组织中ColⅠmRNA的表达（0.41±0.11），有显著性统计学意义（P<0.05或P<0.01）。四磨汤组亦能降低腹腔粘连评分及血清IL-8和TNF-α水平，升高IL-10水平，但与活血通腑方优化方中剂量差异明显。结论：活血通腑方优化方中剂量组可通过调节IL-8，IL-10和TNF-α的水平抑制炎症反应，同时减少细胞外基质ColⅠmRNA的沉积，有效减轻腹腔粘连的程度。

Abstract:

Objective: To observe the therapeutic effect of optimized Huoxue Tongfu (OHT) formula for peritoneal adhesion, to ensure the optimum dosage. Method: Forty-eight male rats were divided randomly into 6 groups:pseudo-operation group, model group, low dose OHT formula group, medium dose OHT formula group and high dose (OHT) formula group and Simo decoction group. All the rats were molded into peritoneal adhesion by file except those in pseudo-operation group. We observed and estimated the peritoneal adhesion and general situation of rats, Elisa method was used to measure the serum level of interleukin (IL)-8, IL-10 and tumor necrosis factor-α (TNF-α), and reverse transcription-polymerase chain reaction for the expression of collagen Ⅰ (Col-Ⅰ) mRNA in adhesional tissue. Result: Compared to pseudo-operation group, in the model group all layers of the intestinal wall showed vessel dilatation, edema, serous layer obviously lymphocyte infiltration, hyperplasia of fibroblasts. The muscular layer structure were morphologically complete, the placenta percreta were slightly thickened and no obvious hyperplasia were found in the fibrous connective tissue in placenta percreta of all the other groups. Compared with the model group, the medium dosage group of optimized Huoxue Tongfu formula, serum IL-8 and TNF-α levels were (154.2±16.2), (58.2±6.4) ng·L^-1, but the level of IL-10 (58.2±6.4) ng·L^-1increased, while decreased expression of Col-ⅠmRNA adhesion tissues (0.41±0.11), there was significant statistical significance (P<0.05 or P<0.01). Simo decoction group could also reduce abdominal adhesion score and serum IL-8 and TNF-α levels, elevated IL-10 levels, but compared with the medium dosage group of OHT formula the difference marked (P<0.05). Conclusion: The medium dosage group of OHT formula can suppress inflammation and relieve peritoneal adhesion by regulating the serum level of IL-8, IL-10 and TNF-α, reducing the deposition of the Col-ⅠmRNA of extracellular matrix.