补中益气颗粒对自身免疫性甲状腺炎模型大鼠小肠肠黏膜超微结构的影响

目的通过观察补中益气颗粒对实验性自身免疫性甲状腺炎(Experimental autoimmune thyroiditis,EAT)模型大鼠小肠肠黏膜超微结构的影响,探讨其改善桥本氏甲状腺炎的作用机制。方法36只Sprague-Dawley(SD)大鼠,随机分为4组,分别为空白组、模型组、补中益气组、硒酵母组4组。除空白组外,其余组大鼠采用高碘喂养法,联合猪免疫球蛋白和弗氏佐剂混合免疫注射法建立EAT大鼠模型。造模后,补中益气组和硒酵母组大鼠分别灌胃补中益气颗粒和硒酵母片。56 d后检测各组大鼠甲状腺过氧化物酶抗体(Thyroid peroxidase antibody,TPOAb)、甲状腺球蛋白抗体(Thyro⁃globulin antibody,TGAb)、血清连蛋白(Zonulin)含量,分离大鼠甲状腺与小肠,在光镜(Light microscope,LM)下观察空白组与模型组甲状腺组织病理变化、各组大鼠小肠组织病理变化,在透射电镜(Transmission electron microscope,TEM)下观察大鼠小肠肠黏膜上皮细胞(Intestinal mucosal epithelial cells,IPC)超微结构变化。结果与空白组比较,模型组血清TPOAb、TGAb、Zonulin含量明显上调(P<0.05)。与模型组比较,补中益气组、硒酵母组血清TPOAb、TGAb、Zonulin含量明显下降(P<0.05)。与空白组比较,LM下模型组大鼠甲状腺组织中有大量淋巴细胞浸润,甲状腺滤泡萎缩、消失,上皮细胞嗜酸性变,小叶间隔增宽,间质纤维化,提示造模成功。EAT大鼠肠绒毛结构紊乱,上皮细胞脱落,固有层裸露,杯状细胞减少,炎性细胞浸润,TEM下IPC微绒毛变短而厚,细胞间隙扩大,紧密连接受损,线粒体肿胀、嵴断裂、基质变淡,细胞器超微结构明显受损。与模型组比较,补中益气组肠绒毛、细胞间紧密连接、细胞器结构受损减轻。结论补中益气汤可以减轻EAT大鼠小肠肠黏膜超微结构损害。

胃肠手术加腹腔感染大鼠肠黏膜超微结构的改变

目的：探讨肠黏膜超微结构改变在胃肠手术加腹腔感染时肠黏膜屏障功能障碍中的意义。方法：采用胃肠手术加腹腔感染大鼠动物模型。随机分为对照组、模型组、胃肠术后1号治疗组。治疗组术后即刻给灌胃胃肠术后1号。对照组和模型组大鼠灌胃等量生理盐水，对胃肠手术加腹腔感染模型大鼠及假手术组大鼠(SO)于术后6、12h取回肠黏膜组织进行光镜及电镜(利用辣根过氧化酶作为黏膜通透性改变的示踪剂)观察。结果：对照组6及12h时相点肠黏膜组织结构基本正常；电镜观察制膜12h肠上皮细胞紧密连接间有高密度电子物质沉着。细胞紧密连接开放；假手术组和胃肠术后1号治疗组肠黏膜上皮细胞紧密连接完整，细胞间隙尢示踪剂沉着。结论：肠黏膜上皮细胞间紧密连接开放，通透性增加是胃肠手术加腹腔感染早期肠黏膜屏障功能障碍的形态学基础。胃肠术后1号能够减轻肠黏膜屏障损害。

大承气汤对烫伤大鼠肠黏膜上皮细胞超微结构变化的影响

目的:了解和揭示大承气汤对SD大鼠烫伤后1周内血浆肿瘤坏死因子、内毒素、肠组织细胞凋亡的影响,观察烫伤大鼠小肠上皮细胞超微结构的变化,探讨本方对肠黏膜屏障保护作用机制。方法:将SD大鼠70只随机分成3组,60只大鼠制备成烫伤模型随机分成中药治疗组(30只)和对照组(30只),中药治疗组给予大承气汤,对照组给予生理盐水灌胃;第3组为正常组10只。治疗组和对照组在给药后第1、3、7天各处死10只,均检测血浆肿瘤坏死因子、内毒素的含量,观察大鼠肠组织细胞凋亡的变化,给药后第7天观察大鼠小肠上皮细胞超微结构变化。结果:大承气汤可明显抑制烫伤大鼠内毒素易位,减少肿瘤坏死因子的产生,抑制肠组织细胞凋亡,明显修复和改善烫伤大鼠肠黏膜细胞超微结构的损伤。结论:中药方剂大承气汤有较理想的肠道屏障保护作用,是预防和治疗肠道屏障功能障碍引发的多器官、多系统功能衰竭的有效的方药。

中药复方便可通片对烫伤大鼠肠黏膜屏障保护作用的实验研究

目的:了解和揭示便可通对SD大鼠烫伤后1周内血浆肿瘤坏死因子、内毒素、肠组织细胞凋亡的影响,观察烫伤大鼠小肠上皮细胞超微结构的变化,探讨本药对肠黏膜屏障保护作用机制。方法:将SD大鼠70只随机分成3组,第1组为中药治疗组(便可通组)30只;第2组为对照组(单纯烫伤组)30只,制备成烫伤模型,给予生理盐水灌胃;第3组为正常组10只。治疗组和对照组在给药后第1、3、7天各处死10只,均检测血浆肿瘤坏死因子、内毒素的含量,观察大鼠肠组织细胞凋亡的变化,给药后第7天观察大鼠小肠上皮细胞超微结构变化。结果:便可通可明显抑制烫伤大鼠内毒素易位,减少肿瘤坏死因子的产生,抑制肠组织细胞凋亡,明显修复和改善烫伤大鼠肠黏膜细胞超微结构的损伤。结论:中医复方便可通有较理想的肠道屏障保护作用,是预防和治疗肠道屏障功能障碍引发的多器官、多系统功能衰竭的有效的药物。

Effect of salvianolate on intestinal epithelium tight junction protein zonula occludens protein 1 in cirrhotic rats

AIM:To study the effect of salvianolate on tight junctions(TJs) and zonula occludens protein 1(ZO-1) in small intestinal mucosa of cirrhotic rats.METHODS:Cirrhosis was induced using carbon tetrachloride.Rats were randomly divided into the untreated group,low-dose salvianolate(12 mg/kg) treatment group,medium-dose salvianolate(24 mg/kg) treatment group,and high-dose salvianolate(48 mg/kg) treatment group,and were treated for 2 wk.Another 10 healthy rats served as the normal control group.Histological changes in liver tissue samples were observed under a light microscope.We evaluated morphologic indices of ileal mucosa including intestinal villi width and thickness of mucosa and intestinal wall using a pathological image analysis system.Ultrastructural changes in small intestinal mucosa were investigated in the five groups using transmission electron microscopy.The changes in ZO-1 expression,a tight junction protein,were analyzed by immunocytochemistry.The staining index was calculated as the product of the staining intensity score and the proportion of positive cells.RESULTS:In the untreated group,hepatocytes showed a disordered arrangement,fatty degeneration was extensive,swelling was obvious,and disorganized lobules were divided by collagen fibers in hepatic tissue,which were partly improved in the salvianolate treated groups.In the untreated group,abundant lymphocytes infiltrated the fibrous tissue with proliferation of bile ducts,and collagen fibers gradually decreased and damaged hepatic lobules were partly repaired following salvianolate treatment.Compared with the untreated group,no differences in intestinal villi width between the five groups were observed.The villi height as well as mucosa and intestinal wall thickness gradually thickened with salvianolate treatment and were significantly shorter in the untreated group compared with those in the salvianolate treatment groups and normal group(P < 0.01).The number of microvilli decreased and showed irregular lengths and arrangements in the untreated group.The intercellular space between epithelial cells was wider.The TJs were discontinuous,which indicated disruption in TJ morphology in the untreated group.In the treated groups,the microvilli in the intestinal epithelium were regular and the TJs were gradually integrated and distinct.The expression of ZO-1 decreased in the small intestine of the untreated cirrhotic rats.The high expression rate of ZO-1 in ileal mucosa in the untreated group was significantly lower than that in the medium-dose salvianolate group(21.43% vs 64.29%,χ 2 = 5.25,P < 0.05),high-dose salvianolate group(21.43% vs 76.92%,χ 2 = 8.315,P < 0.01) and normal group(21.43% vs 90%,χ 2 = 10.98,P < 0.01).CONCLUSION:Salvianolate improves liver histopathological changes,repairs intestinal mucosa and TJ structure,and enhances ZO-1 expression in the small intestinal mucosa in cirrhotic rats.