摘要目的:探讨加味参苓白术散对不同含糖量腹膜透析液（PDF）诱导腹膜纤维化大鼠腹膜组织病理学改变和细胞外基质（ECM）的影响及其机制。方法:将70只雄性SD大鼠按照单纯随机抽样原则分为对照组、不同含糖量PDF模型组及其相应中药干预组，每组10只。分别腹腔注射含糖量为1.5%、2.5%、4.25%的PDF 100 mL·kg -1·d -1诱导大鼠腹膜纤维化模型，每日1次，连续8周；对照组大鼠给予等量生理盐水。各中药干预组大鼠每日于制模后即刻灌胃加味参苓白术散溶液10 mL/kg（每升含2.014 kg生药）；各PDF模型组及对照组大鼠灌胃等量生理盐水。8周后，检测各组大鼠腹膜超滤量。取腹膜组织，苏木素-伊红（HE）染色后，光镜下观察壁层腹膜形态学改变及腹膜厚度；Masson染色后，光镜下观察腹膜胶原纤维沉积情况；采用蛋白质免疫印迹试验（Western blotting）测定腹膜组织中ECM主要成分E-钙黏蛋白（E-cadherin）、α-平滑肌肌动蛋白（α-SMA）和波形蛋白（Vimentin）的表达；采用免疫组化法检测ECM调控因子基质金属蛋白酶-9（MMP-9）、金属蛋白酶组织抑制剂-1（TIMP-1）和转化生长因子-β1（TGF-β1）的阳性表达。 结果:与对照组比较，不同含糖量PDF均可诱导大鼠发生腹膜纤维化，且纤维化程度随含糖量增加而逐渐加重，表现为腹膜增厚，胶原纤维沉积增加，腹膜超滤量减少，伴随腹膜组织中E-cadherin、MMP-9表达下调，α-SMA、Vimentin、TIMP-1、TGF-β1表达上调，以4.25% PDF模型组腹膜组织病理学改变及ECM聚积情况更加严重。经加味参苓白术散干预后，与相应PDF模型组比较，大鼠腹膜纤维化均得到不同程度改善，仍以4.25% PDF相应中药干预组效果更为显著，壁层腹膜明显变薄（μm：101.86±16.01比140.65±10.13， P<0.05），胶原纤维沉积明显减少，腹膜超滤量明显增加（mL：-0.01±3.45比-3.53±1.84， P<0.05），同时腹膜组织中E-cadherin、MMP-9表达明显上调〔E-cadherin蛋白（E-cadherin/β-actin）：0.84±0.08比0.28±0.05，MMP-9（ A值）：0.60±0.15比0.37±0.01，均 P<0.05〕，α-SMA、Vimentin、TIMP-1、TGF-β1表达明显下调〔α-SMA蛋白（α-SMA/β-actin）：0.36±0.08比1.05±0.09，Vimentin蛋白（Vimentin/β-actin）：0.53±0.07比1.19±0.04，TIMP-1（ A值）：0.49±0.06比0.87±0.02，TGF-β1（ A值）：0.67±0.04比0.89±0.10，均 P<0.05〕。 结论:大鼠腹膜纤维化程度随PDF含糖量增加而逐渐加重；加味参苓白术散对不同含糖量PDF诱导的大鼠腹膜纤维化均有一定改善作用，其机制与调节纤维化标志物表达及减少ECM聚积有关。

abstractsObjective:To investigate the effect of modified Shenling Baizhu San on the pathological changes and extracellular matrix (ECM) in rats with peritoneal fibrosis induced by peritoneal dialysate fluid (PDF) with different sugar concentration and its mechanism.Methods:Seventy male Sprague-Dawley (SD) rats were randomly divided into control group, different sugar content PDF model groups and corresponding traditional Chinese medicine intervention groups, with 10 rats in each group. Peritoneal fibrosis model was reproduced by intraperitoneal injection of 100 mL·kg -1·d -1 PDF containing 1.5%, 2.5% and 4.25% sugar once a day for 8 weeks. The rats in the control group were given the same amount of normal saline. The rats in the traditional Chinese medicine intervention groups were treated with gavage of 10 mL/kg of modified Shenling Baizhu San (containing 2.014 g crude drug per liter) immediately after modeling. The PDF model groups and the control group were given the same amount of normal saline by gavage. After 8 weeks, the peritoneal ultrafiltration volume of rats in each group was measured. The peritoneal tissues were collected and stained with hematoxylin-eosin (HE), and the structural changes and thickness of the parietal peritoneum were observed under a light microscope. After Masson staining, the deposition of collagen fibers was observed under a light microscope. Western blotting was used to detect the protein expressions of E-cadherin,?α-smooth muscle actin (α-SMA) and Vimentin, the main components of ECM in parietal peritoneum. The positive expressions of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and transforming growth factor-β1 (TGF-β1) were detected by immunohistochemical staining. Results:Compared with the control group, PDF with different sugar contents could induce peritoneal fibrosis in rats, and the degree of fibrosis was gradually aggravated with the increase of sugar content, which was manifested as peritoneal thickening, increased collagen fiber deposition, decreased peritoneal ultrafiltration volume, down-regulated expressions of E-cadherin and MMP-9 in peritoneal tissue, and up-regulated expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1, and the pathological changes and ECM accumulation in peritoneal tissues were more serious in 4.25% PDF model group. After the intervention of modified Shenling Baizhu San, compared with the corresponding PDF model groups, the peritoneal fibrosis of rats was improved to varying degrees, and the effect of the 4.25% PDF+traditional Chinese medicine intervention group was more significant, the parietal peritoneum was significantly thinner (μm: 101.86±16.01 vs. 140.65±10.13, P < 0.05), collagen fiber deposition was significantly reduced, peritoneal ultrafiltration volume was significantly increased (mL: -0.01±3.45 vs. -3.53±1.84, P < 0.05), the expressions of E-cadherin and MMP-9 in peritoneal tissues were significantly up-regulated [E-cadherin protein (E-cadherin/β-actin): 0.84±0.08 vs. 0.28±0.05, MMP-9 ( A value): 0.60±0.15 vs. 0.37±0.01, both P < 0.05], and the expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1 were significantly down-regulated [α-SMA protein (α-SMA/β-actin): 0.36±0.08 vs. 1.05±0.09, Vimentin protein (Vimentin/β-actin): 0.53±0.07 vs. 1.19±0.04, TIMP-1 ( A value): 0.49±0.06 vs. 0.87±0.02, TGF-β1 ( A value): 0.67±0.04 vs. 0.89±0.10, all P < 0.05]. Conclusions:The degree of peritoneal fibrosis gradually increased with the increase of PDF sugar content in rats. Modified Shenling Baizhu San can improve peritoneal fibrosis induced by PDF with different sugar contents in rats, and the mechanism is related to the changes in the expression of fibrosis markers and ECM accumulation.