目的:通过观察全反式维甲酸对慢性酒精性肝损伤大鼠肝脏TGF-β1、CTGF和Collal表达的影响,探讨该药物对酒精性肝纤维化形成的作用．方法:大鼠24只随机分为3组:酒精组(J组),给予酒精-玉米油混悬液灌胃;治疗组(A组),给予上述混悬液灌胃8 w...目的:通过观察全反式维甲酸对慢性酒精性肝损伤大鼠肝脏TGF-β1、CTGF和Collal表达的影响,探讨该药物对酒精性肝纤维化形成的作用．方法:大鼠24只随机分为3组:酒精组(J组),给予酒精-玉米油混悬液灌胃;治疗组(A组),给予上述混悬液灌胃8 wk后加用0,15 mg／(kg·d) 的全反式维甲酸灌胃;对照组(N组),给予等量的生理盐水和玉米油灌胃．16 wk后处死大鼠．光、电镜下观察肝组织病理改变,高压液相色谱法(HPLC)测肝组织中维甲酸的含量,免疫组化法检测肝组织中转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)的蛋白水平,逆转录聚合酶链反应(RT-PCR)检测肝组织中TGF-β1、CTGF和Ⅰ型胶原前胶原α1(Collal)的mRNA水平．结果:光镜下酒精组及治疗组均呈不同程度的酒精性肝炎改变,电镜下酒精组肝细胞线粒体肿胀,内质网扩张,脱颗粒,而治疗组改变轻于酒精组．肝组织中维甲酸含量:酒精组低于正常组,治疗组接近对照组水平．Collal 的mRNA表达在酒精组中较对照组明显增高 (0．18±0．03 vs 0．10±0．02,P<0．01),治疗组较酒精组表达下降(0．14±0．03 vs 0．18±0．03,P <0．05)．TGF-β1的mRNA及蛋白表达在酒精组中增高(0．53±0．17 vs 0．34±0．05,105．93 ±10．12 vs 149．27±10．17,P<0．01),治疗组相对于酒精组有所下降(0．41±0．06 vs 0．53± 0．17,130．80±6.23 vs 105．93±10．12,P<0．05)． CTGF的mRNA及蛋白表达在酒精组中增高 (0．41±0．13 vs 0．17±0．05,130．84±5．72 vs 158.37±6．64,P<0．05),治疗组对于酒精组有所下降(0.30±0．04 vs 0．41±0．13,149．23± 6．65 vs 130．84±5．72．P<0．05)．结论:小剂量全反式维甲酸通过降低慢性酒精性肝损伤大鼠肝脏致纤维化因子TGF-β1, CTGF和Collal的表达抑制早期酒精性肝纤维化的形成．展开更多
克罗恩病（CD）属于特发性肠道慢性炎性疾病，其发病机制不明，传统的治疗药物（包括氨基水杨酸、糖皮质激素和免疫抑制剂等）长期应用于临床，有一定疗效，但对疾病的自然病程改善不大。近年研究发现，细胞因子在 CD 患者肠道炎性反应...克罗恩病（CD）属于特发性肠道慢性炎性疾病，其发病机制不明，传统的治疗药物（包括氨基水杨酸、糖皮质激素和免疫抑制剂等）长期应用于临床，有一定疗效，但对疾病的自然病程改善不大。近年研究发现，细胞因子在 CD 患者肠道炎性反应的发生、发展中发挥了重要的作用，阻断细胞因子效应可有效缓解肠道炎性反应。国外已研发出针对不同细胞因子的多种生物制剂，目前最为成熟并已在国内批准应用于 CD 的生物制剂是英夫利西单抗（IFX）。IFX 是一种能与肿瘤坏死因子-α（TNF-α）特异性结合的人-鼠嵌合型免疫球蛋白 IgG1单克隆抗体，具有75％人源性和25％鼠源性，可与 TNF-α的可溶形式和跨膜形式以高亲和力结合，抑制TNF-α与其受体结合，降低 TNF-α的生物学活性，从而降低血清中白细胞介素-6（IL-6）和 C-反应蛋白（CRP）水平，迅速缓解肠道炎性反应［1］。本科室自2009年至今已应用 IFX 治疗不同类型 CD 患者40余例，部分患者疗效显著，但考虑到长期应用的不良反应及费用，如何合理应用该药物成为值得关注的问题。本文就 IFX 的适用患者、用药时机及方法，结合个人诊治体会进行述评。展开更多
Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. The aim of the study was to use this experimental model to elucidate if Thl 7 cells are involve...Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. The aim of the study was to use this experimental model to elucidate if Thl 7 cells are involved in the development of gastrointestinal hypermotility. Colonic smooth muscle contractility was investigated in response to acetylcholine. The levels of IL-17, IL-23 and TGF-β1 in colon were measured by Western blotting. Flow cytometric detection of intracellular IFN-7/IL-4/ IL- 17 cytokine production was used to analyze the proportions of CD4＋ T cells subsets in colon. Our results showed that colonic muscle contractility was increased 2 weeks post infection （PI） and stayed high 12 weeks PI when no discernible inflammation was present in the gut. The proportion of Th17 cells and the expression of IL-17 were up-regulated in colon 2 weeks PI and returned to normal 8 weeks PI. The content of IL-17 was correlated with the colonic smooth muscle hypercontracility 2 weeks PI. Meanwhile, TGF-β1 was increased 2 weeks PI, while IL-23 was normal. Our results suggest that Th17 cells affect the colonic muscle contractility in mice infected with Trichinella spiralis at intestine stage but not at muscle stage and the effect of Th17 cells on muscle contractility might be induced by TGF-β1. Other cytokines might be involved in the hypercontracility of colonic smooth muscle at muscle stage.展开更多
Toll-like receptors （TLRs） are key components of the innate immune system which trigger antimicrobial host defense responses. This study aimed to investigate the impact of probiotics （Lactoba- cillus, Bifiidobacter...Toll-like receptors （TLRs） are key components of the innate immune system which trigger antimicrobial host defense responses. This study aimed to investigate the impact of probiotics （Lactoba- cillus, Bifiidobacterium） on the expression of TLR4 and tumor necrosis factor-alpha （TNF-α in the co- lon mucosa of rat experimental ulcerative colitis model induced by trinitrobenzene sulfonic acid （TNBS）/ethanol and immune complexes. The gross and histological changes of the colonic mucosa were observed and assessed by the means-standard deviation and independent samples t-test. The pro- tein expression levels of TLR4 and TNF-α were detected by using immunohistochemistry and Westem blotting, respectively. It was revealed that there was visible infiltration of inflammatory cells, formation of crypt abscess, and the reduction of goblet cells in the colon tissue of experimental models. As com- pared with the control group, the levels of TLR4 and TNF-α protein were significantly increased in the model group （P〈0.01 for both）. No significant difference was found in the expression of TLR4 and TNF-α between the two-week probiotics treatment group and the model group （P〉0.05）, whereas sig- nificant reductions were shown in rats which were treated with probiotics for four weeks as compared with the model group （P〈0.01）. There was no significant difference between two probiotics-treated groups. Our results implied that probiotics were likely to play a key role in protecting ulcerative colitis by reducing the inflammatory factor TNF-α expression through inhibiting the TLR4 expression in the colon tissue of experimental models.展开更多
The effects of all-trans-retinoic acid （ATRA） in low doses supplementation on concentrations of polar retinoid metabolites （PRM） and retinoids in the ethanol-fed rat liver, and on hepatocyte injury were investigat...The effects of all-trans-retinoic acid （ATRA） in low doses supplementation on concentrations of polar retinoid metabolites （PRM） and retinoids in the ethanol-fed rat liver, and on hepatocyte injury were investigated. The rat model of alcoholic liver disease （ALD） was induced by intragastric infusion of ethanol, and then the rats were administrated with ATRA in two different doses （150 μg/kg body weight and 1.5 mg/kg body weight） for 4 weeks. Concentrations of retinoids in rat liver and plasma were determined by using HPLC. Liver tissues pathologic changes were observed under the light microscopy and electron microscopy. The serum transaminases concentrations were measured. The results showed that the HPLC analysis of retinoids revealed that retinoids （vitamin A, RA, retinyl palmitate） concentrations in ethanol-fed rat liver and RA concentration in ethanol-fed rat plasma were markedly diminished （P〈0.0 l） after ethanol feeding for 12 weeks. Furthermore, obvious peaks of PRM were formed in livers of ethanol-fed rats. ATRA 150 [tg/kg supplementation in ethanol-fed rats for 4 weeks raised RA concentration in both liver and plasma, and also raised vitamin A concentration in liver to control levels, partially restored retinyl palmitate concentration （P〈0.05） in liver. ATRA 1.5 mg/kg supplementation raised not only RA concentrations in liver and plasma but also retinyl palmitate concentrations in liver. However, the vitamin A concentration in liver of ATRA-supplemented rats （ 1.5 mg/kg） was higher than that of controls （P〈0.05）. The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling, steatosis, the swelling of mitochondria and proliferation of smooth endoplasmic reticulum （SER）. ATRA treatment greatly decreased levels of serum transaminases as compared with the only ethanol-fed group （P〈0.05）. It was concluded that low-dose ATRA treatment could restore retinoids concentrations and abolish the PRM formation in liver of ALD rats, and then ameliorate the injury of liver cells.展开更多
文摘目的:通过观察全反式维甲酸对慢性酒精性肝损伤大鼠肝脏TGF-β1、CTGF和Collal表达的影响,探讨该药物对酒精性肝纤维化形成的作用．方法:大鼠24只随机分为3组:酒精组(J组),给予酒精-玉米油混悬液灌胃;治疗组(A组),给予上述混悬液灌胃8 wk后加用0,15 mg／(kg·d) 的全反式维甲酸灌胃;对照组(N组),给予等量的生理盐水和玉米油灌胃．16 wk后处死大鼠．光、电镜下观察肝组织病理改变,高压液相色谱法(HPLC)测肝组织中维甲酸的含量,免疫组化法检测肝组织中转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)的蛋白水平,逆转录聚合酶链反应(RT-PCR)检测肝组织中TGF-β1、CTGF和Ⅰ型胶原前胶原α1(Collal)的mRNA水平．结果:光镜下酒精组及治疗组均呈不同程度的酒精性肝炎改变,电镜下酒精组肝细胞线粒体肿胀,内质网扩张,脱颗粒,而治疗组改变轻于酒精组．肝组织中维甲酸含量:酒精组低于正常组,治疗组接近对照组水平．Collal 的mRNA表达在酒精组中较对照组明显增高 (0．18±0．03 vs 0．10±0．02,P<0．01),治疗组较酒精组表达下降(0．14±0．03 vs 0．18±0．03,P <0．05)．TGF-β1的mRNA及蛋白表达在酒精组中增高(0．53±0．17 vs 0．34±0．05,105．93 ±10．12 vs 149．27±10．17,P<0．01),治疗组相对于酒精组有所下降(0．41±0．06 vs 0．53± 0．17,130．80±6.23 vs 105．93±10．12,P<0．05)． CTGF的mRNA及蛋白表达在酒精组中增高 (0．41±0．13 vs 0．17±0．05,130．84±5．72 vs 158.37±6．64,P<0．05),治疗组对于酒精组有所下降(0.30±0．04 vs 0．41±0．13,149．23± 6．65 vs 130．84±5．72．P<0．05)．结论:小剂量全反式维甲酸通过降低慢性酒精性肝损伤大鼠肝脏致纤维化因子TGF-β1, CTGF和Collal的表达抑制早期酒精性肝纤维化的形成．
基金supported by a grant from the National Natural Sciences Foundation of China(No.30770986)
文摘Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. The aim of the study was to use this experimental model to elucidate if Thl 7 cells are involved in the development of gastrointestinal hypermotility. Colonic smooth muscle contractility was investigated in response to acetylcholine. The levels of IL-17, IL-23 and TGF-β1 in colon were measured by Western blotting. Flow cytometric detection of intracellular IFN-7/IL-4/ IL- 17 cytokine production was used to analyze the proportions of CD4＋ T cells subsets in colon. Our results showed that colonic muscle contractility was increased 2 weeks post infection （PI） and stayed high 12 weeks PI when no discernible inflammation was present in the gut. The proportion of Th17 cells and the expression of IL-17 were up-regulated in colon 2 weeks PI and returned to normal 8 weeks PI. The content of IL-17 was correlated with the colonic smooth muscle hypercontracility 2 weeks PI. Meanwhile, TGF-β1 was increased 2 weeks PI, while IL-23 was normal. Our results suggest that Th17 cells affect the colonic muscle contractility in mice infected with Trichinella spiralis at intestine stage but not at muscle stage and the effect of Th17 cells on muscle contractility might be induced by TGF-β1. Other cytokines might be involved in the hypercontracility of colonic smooth muscle at muscle stage.
基金supported the grants from the Natural Science Foundation of Hubei province(No.2012FFB02325)the National Natural Science Foundation of China(No.81000159)
文摘Toll-like receptors （TLRs） are key components of the innate immune system which trigger antimicrobial host defense responses. This study aimed to investigate the impact of probiotics （Lactoba- cillus, Bifiidobacterium） on the expression of TLR4 and tumor necrosis factor-alpha （TNF-α in the co- lon mucosa of rat experimental ulcerative colitis model induced by trinitrobenzene sulfonic acid （TNBS）/ethanol and immune complexes. The gross and histological changes of the colonic mucosa were observed and assessed by the means-standard deviation and independent samples t-test. The pro- tein expression levels of TLR4 and TNF-α were detected by using immunohistochemistry and Westem blotting, respectively. It was revealed that there was visible infiltration of inflammatory cells, formation of crypt abscess, and the reduction of goblet cells in the colon tissue of experimental models. As com- pared with the control group, the levels of TLR4 and TNF-α protein were significantly increased in the model group （P〈0.01 for both）. No significant difference was found in the expression of TLR4 and TNF-α between the two-week probiotics treatment group and the model group （P〉0.05）, whereas sig- nificant reductions were shown in rats which were treated with probiotics for four weeks as compared with the model group （P〈0.01）. There was no significant difference between two probiotics-treated groups. Our results implied that probiotics were likely to play a key role in protecting ulcerative colitis by reducing the inflammatory factor TNF-α expression through inhibiting the TLR4 expression in the colon tissue of experimental models.
文摘The effects of all-trans-retinoic acid （ATRA） in low doses supplementation on concentrations of polar retinoid metabolites （PRM） and retinoids in the ethanol-fed rat liver, and on hepatocyte injury were investigated. The rat model of alcoholic liver disease （ALD） was induced by intragastric infusion of ethanol, and then the rats were administrated with ATRA in two different doses （150 μg/kg body weight and 1.5 mg/kg body weight） for 4 weeks. Concentrations of retinoids in rat liver and plasma were determined by using HPLC. Liver tissues pathologic changes were observed under the light microscopy and electron microscopy. The serum transaminases concentrations were measured. The results showed that the HPLC analysis of retinoids revealed that retinoids （vitamin A, RA, retinyl palmitate） concentrations in ethanol-fed rat liver and RA concentration in ethanol-fed rat plasma were markedly diminished （P〈0.0 l） after ethanol feeding for 12 weeks. Furthermore, obvious peaks of PRM were formed in livers of ethanol-fed rats. ATRA 150 [tg/kg supplementation in ethanol-fed rats for 4 weeks raised RA concentration in both liver and plasma, and also raised vitamin A concentration in liver to control levels, partially restored retinyl palmitate concentration （P〈0.05） in liver. ATRA 1.5 mg/kg supplementation raised not only RA concentrations in liver and plasma but also retinyl palmitate concentrations in liver. However, the vitamin A concentration in liver of ATRA-supplemented rats （ 1.5 mg/kg） was higher than that of controls （P〈0.05）. The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling, steatosis, the swelling of mitochondria and proliferation of smooth endoplasmic reticulum （SER）. ATRA treatment greatly decreased levels of serum transaminases as compared with the only ethanol-fed group （P〈0.05）. It was concluded that low-dose ATRA treatment could restore retinoids concentrations and abolish the PRM formation in liver of ALD rats, and then ameliorate the injury of liver cells.
