复方蜚蠊提取物缓解噁唑酮诱导大鼠活动期溃疡性结肠炎的作用机制

目的 研究复方蜚蠊提取物Ento-PB对噁唑酮诱导的活动期溃疡性结肠炎(UC)大鼠的治疗作用,并初步探讨其可能作用机制.方法 采用噁唑酮诱导的溃疡性结肠炎大鼠模型,灌胃给予柳氮磺胺吡啶(SASP,0.3g/kg)和复方Ento-PB (0.05、0.1、0.2 g/kg),通过测定大鼠DAI (疾病活动指数)、CMDI...     

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??OBJECTIVE To investigate the protective effect and possible mechanism of Rhizoma Coptis(RC) on lipopolysaccharide(LPS)-induced inflammatory injury in rat hepatocytes(BRL). METHODS LPS-induced BRL cells injury model was established in vitro, then the damaged cells were given different interventions and treatment with 0.175, 0.1 mg?? mL^-1 RC aqueous extract as the test drug, and dexamethasone(Dex) as positive control drug. The optimal test doses of LPS and RC aqueous extract were selected and determined by cell counting kit-8(CCK-8), the cellular apoptosis rate was determined by flow cytometry, TLR4/NF-??B and TLR4/IRF3 signaling pathways and the mRNA level of related inflammatory mediators(TNF-??, IL-1??, IL-6) were detected by RT-PCR, the NF-??B p65 protein expression was analysed by Western blot and immunofluorescence techniques. RESULTS ??Compared with normal control group, 0.1 mg??mL^-1 LPS affected on BRL cells for 24 h, the cell survival rate was decreased significantly(P<0.01), the apoptotic rate increased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 were significantly increased(P<0.01), and the NF-??B p65 protein expression was increased. ??Compared with the model group, 0.1 and 0.175 mg??mL^-1 RC affected on LPS-induced BRL cells for 24 h, the survival rate of BRL cells was increased significantly(P<0.05), the apoptotic rate decreased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 and the NF-??B p65 protein expression were decreased significantly(P<0.01). CONCLUSION Rhizoma Coptis has obviously protective effect on LPS-induced inflammatory injury in rat hepatocytes(BRL), the mechanism of which may be related with inhibiting apoptosis, reducing the release of inflammatory factors such as TNF-????IL-1?? and IL-6, blocking NF-??B p65 protein nuclear translocation, interfering the R4/NF-??B and TLR4/IRF3 signaling pathway.     

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??OBJECTIVE To investigate the anti-inflammatory effects of extract of Scutellaria baicalensis Georgi (SGE) and underlying mechanism by using LPS-induced microglial BV2 cells. METHODS MTT assay was used to observe the cell viability. The content of NO in cell supernatant was measured by Griess reagent. The levels of IL-1??, IL-6 and TNF-?? were detected by ELISA kits. The intracellular TLR4 expression was assayed by Western blotting. RESULTS The levels of NO, IL-1??, IL-6 and TNF-?? were significantly increased induced by LPS in the supernatant of BV2 cells (all P<0.01). However, co-treatment with SGE 100 ??g??mL^-1 significantly decreased the production of related inflammatory factors including NO (P<0.01), IL-1??(P<0.01), IL-6 (P<0.01) and TNF-?? (P<0.05). Furthermore, SGE significantly inhibited the TLR4 expression induced by LPS in BV2 cells. CONCLUSION SGE is able to alleviate LPS-induced inflammatory responses in BV2 cells through down-regulation of TLR4 protein expression suggesting that SGE has therapeutic potential for the treatment of neuroinflammatory diseases.     

肠炎清对溃疡性结肠炎模型大鼠中分泌型免疫球蛋白A和P选择素的影响

目的： 探讨肠炎清对免疫复合法诱致溃疡性结肠炎模型大鼠结肠组织中分泌型免疫球蛋白A（sIgA）和P选择素的影响研究。 方法： SD大鼠64只,雌雄各半,随机分为正常对照组（12只）和模型组（52只）;模型组应用免疫复合法（注射抗原乳化剂+TNBS/乙醇）建立溃疡性结肠炎大鼠模型,正常组注射生理盐水。再喂养3周后,随机从造模组中抽取4只处死,检查结肠病变情况,确定溃疡性结肠炎模型建立;随后将造模大鼠随机分为4组：肠炎清低、高剂量组（10,40 g·kg^-1）,柳氮磺吡啶（SASP,0.1 g·kg^-1）组和模型对照组;每组12只,每天灌胃给药1次,连续给药4周。每周观察大鼠疾病活动指数（DAI）;末次给药后,处死动物,解剖观察并评分结肠黏膜损伤指数（CMDI）;光镜下观察结肠组织的病理学变化,并做组织病理变化评分（HS）;采用ELISA方法测定结肠组织中sIgA的含量变化;采用免疫组化法（SP法）检测P选择素的表达变化。 结果： 与正常组比较,模型大鼠的DAI,CMDI,结肠HS评分和髓过氧化物酶（MPO）均明显上升（P<0.01）;结肠组织中的sIgA含量显著下降,而P选择素表达明显增加 （P<0.01）;与模型组比较,肠炎清治疗4周后大鼠的DAI,CMDI,结肠HS评分和MPO值均能显著下降（P<0.01）;结肠组织中sIgA的浓度升高,同时减少了P选择素表达（P<0.01）。 结论： 肠炎清对免疫复合法诱致溃疡性结肠炎模型大鼠具有治疗作用,其机制可能与提升结肠组织内sIgA浓度和降低P选择素的表达有关。     

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??OBJECTIVE To investigate the effect of ??-secretase inhibitor DAPT in inflammation-induced brain white matter injury in neonatal mice.METHODS Sixty C57BL/10J neonatal mice are randomly divided into control group, control+DAPT (10 mg??kg^-1) group, inflammation (LPS) group, LPS+DAPT group (inflammation exposure after 10 mg??kg^-1 DAPT treatment). All neonatal mice were killed and brain was removed to the following observation and detection:at P5, the mRNA expression variation of IL-1??, IL-8,TNF-??,Hes1 and NICD by Real-time PCR methods. Oligodendrocytes were identified by immunofluorescence staining. Myelin basic protein (MBP) protein expression was detected by Western blot assay.RESULTS LPS group showed brain injury characterized by inhibition of brain development. There were significant differences in mRNA expression of IL-1??, IL-8, TNF-??, Hes1 and NICD between LPS+DAPT group and LPS group (P<0.05), and the mRNA expression of IL-1??, IL-8,TNF-??,Hes1 and NICD in inflammation-treated were significantly increased than control group (P<0.05). The results showed more expression of MBP in LPS+DAPT group compared with LPS group (P<0.05). Compared with the blank control group, which was obviously decreased after 48 h of inflammation (P<0.05).CONCLUSION Inflammation leads to abnormal of notch signal expression in neonatal mice, and which is shows inflammation involved in brain damage.Its mechanism is probably associated with the maturation of oligodendrocytes.     

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??OBJECTIVE To clarify the effect of total glucosides from paeony(PTG) on irritable bowel syndrome (IBS), and to explore the molecular mechanism of PTG on alleviating diarrhea symptoms and abdominal pain.METHODS The diarrhea model was conducted by exposing rat to restraint stress stimulation and bellyache model was conducted by subcutaneous injection of neostigmine to mice. Based on these two models, the curative effect of PTG on IBS was investigated. To investigate the regulative effects of PTG on Caco-2 cells, the Caco-2 monolayer cell model with barrier dysfunction was established by trypsin stimulation, and the inflammatory Caco-2 cell model was established by interleukin-1?? (IL-1??) stimulation.RESULTS PTG could significantly reduce the frequency of defecation in diarrhea rat model (P<0.05) and relieve abnormal bowel movements in bellyache mice model (P<0.05). After PTG treatment, the TEER value of Caco-2 monolayer was significantly increased (P<0.01), the transmittance of fluorescence yellow was significantly decreased (P<0.001) and the expression of tight junction (ZO-1)protein was notably up-regulated (P<0.001). In addition, the gene and protein expression of nuclear factor profilin kappa B(I??B??)in inflammatory Caco-2 cell model was significantly improved (P<0.001) after PTG treatment.CONCLUSION PTG significantly ameliorates IBS symptoms by protecting the barrier function of Caco-2 cell monolayer and relieving inflammation of Caco-2 cells.     

清肠温中方对三硝基苯磺酸诱导的实验性结肠炎大鼠肠黏膜作用的研究

目的研究清肠温中方对三硝基苯磺酸诱导的实验性结肠炎大鼠结肠黏膜的作用。方法以三硝基苯磺酸灌肠诱导大鼠急性结肠炎,造模成功后连续以清肠温中方高、中、低剂量混悬液及柳氮磺吡啶（salazosulfapyridine,SASP）混悬液灌胃治疗10天,取结肠,进行疾病活动指数（disease activity index,DAI）,结肠黏膜损伤指数（colonic mucosa damageindex,CMDI）和结肠组织病理学（histological score,HS）评价。结果药物干预10天后,模型组DAI、CMDI、HS明显高于正常组（P〈0．05）;SASP组DAI及CMDI均较模型组有明显的下降（P〈0．05）;清肠温中方各剂量组DAI、CMDI、HS水平较模型组均明显下降（P〈0．05）,其中清肠温中方高、中剂量组炎细胞侵润的程度明显低于模型组（P〈0．05）。结论清肠温中方对三硝基苯磺酸诱导的结肠炎大鼠肠黏膜有保护作用,且能有效改善结肠组织炎细胞侵润。     

康复新液对噁唑酮诱导溃疡性结肠炎大鼠治疗作用及机制初探

目的:研究康复新液对噁唑酮(oxazolone,OXZ)诱导溃疡性结肠炎(ulcerative colitis,UC)大鼠的治疗效果,并初步探讨其机制。方法:将SD大鼠分为正常组、模型组、美沙拉嗪组和康复新低、中、高剂量组,后5组以OXZ溶液灌肠诱导UC模型。造模后第1天开始各组灌肠给药,每天1次,连续给药7 d。每天进行疾病活动指数(disease activity index,DAI)评分,造模后第8天处死大鼠,进行结肠黏膜损伤指数(colonmucosa damage index,CMDI)评分和病理组织学评分(histopathological score,HS);采用酶联免疫吸附测定方法检测血清中白细胞介素-4(IL-4)含量和结肠黏膜中IL-13含量。结果:模型组大鼠的DAI评分,结肠指数,CMDI评分,HS和结肠黏膜中IL-13含量均显著高于正常组(P0.05,P0.01),而模型组大鼠血清中IL-4含量显著低于正常组(P0.01)。与模型组比较,康复新高剂量能显著降低大鼠DAI评分,CMDI评分,HS和结肠黏膜中IL-13含量(P0.05,P0.01),而血清中IL-4含量显著升高(P0.01)。结论:康复新液能够有效地缓解噁唑酮诱导的溃疡性结肠炎。     

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??OBJECTIVE To observe the effect of chlorogenic acid on the secretion of inflammatory cytokines and regulation mechanism of P38MAPK, NF-??B signaling pathway in human hepatic stellate cells induced by TGF-??1. METHODS Different concentrations of CGA worked in normal and activated hepatic stellate cells to make sure the appropriate drug concentration.The exponential growth phase cells were randomly divided into normal HSC group, normal HSC+CGA group, after cultured 48 h, the cells were cultured with 0??50??100 mg??L^-1 CGA for 24 h; HSC(TGF-??1) group?? HSC(TGF-??1 + CGA) group: after 24 h, the cells were induced by 10 ??g??L^-1 TGF-??1 for 24 h, and then cultured with 0, 50, 100 mg??L^-1 CGA for 24 h. The expression of ??-SMA protein was detected by immunocytochemistry, the expression of p-P38, P65 protein was detected by Western-blot, the expression of TNF-??, IL-6 mRNA was detected by real time quantitative PCR, and the content of TNF-??, IL-6 in the supernatant was detected by ELISA method. RESULTS The appropriate concentrations of CGA were 50 and 100 mg??L^-1, these concentration has no effect on normal HSC(P>0.05); after stimulation by TGF-??1, the expression of ??-SMA, p-P38, P65, TNF-??, IL-6 was increased(P<0.01), when activated HSC cells were treated with 50 and 100 mg??L^-1 CGA, the expression of ??-SMA, p-P38, P65, TNF-??, IL-6 was decreased(P<0.05, P<0.01). CONCLUSION CGA can inhibit the proliferation of activated HSC, regulate the secretion of inflammatory factors such as TNF-??, IL-6 by P38MAPK and NF-??B signaling pathway, inhibit the occurrence of liver fibrosis.     

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??OBJECTIVE To explore the expression of miR-199a in ulcerative colitis (UC) rats induced by 2,4,6-trinitrobenzene sulfonic (TNBS) / ethanol and the study on the effect of TWP on them. METHODS Through injecting TNBS/ethyl alcohol acid liquid into the anus of the rats to establish the UC rat model . The colitics commom morphous damage and grade the histopathological score (CMDI) of colon mucosa injury were evaluated. Chip analysis and Real-time PCR were used to verify the expression of miR-199a in each colon mucosa tissue. Based on the expression profile, the downstream target genes mRNA in milwalk database was selected, then the expression of target genes mRNA by Real-time PCR in each group was veritied, at last the relevant signal pathway in the DAVID database was analyzed. Doing these to analyse the target gene mRNA regulated by the miR-199a in the inflammatory activity of UC. RESULTS Compared with the model group, TWP high dose group was significantly lower on gross morphological damage score and histopathological injury score(P??0.01). Chip analysis showed that in model group, the expression of miR-199a was significantly higher than the normal group(P??0.01), and expression of the AZA group was significantly lower than the model group(P??0.01,P??0.05). The expression of miR-199a-3p in medium dose group and the expression of miR-199a-5p in high dose group were significantly lower than the model group(P??0.05).The RESULTS of Real-time PCR showed that expression of miR-199a in the model group was significantly increased than that in the normal group(P??0.01). The expression of miR-199a-3p in TWP medium dose group, high dose group and AZA group were decreased than that in model group(P??0.05). Meanwhile, the expression of miR-199a-5p in TWP medium dose group was decreased than that in model group(P??0.05). The gene expression profile showed that FASL was the target gene of miR-199a. In the model group, the expression of FASL was higher than that in the normal group. The expression of FASL in AZA group was significantly decreased than that in the model group(P??0.01). The RESULTS by the Real-time PCR of the target gene FASL showed that in the model group, the expression of FASL was higher than that in the normal group (P< 0.01). The expression of FASL in medium dose group, high dose group and AZA group were significantly decreased than that in the model group (P<0.01). CONCLUSION miR-199a is up-regulated in TNBS/Ethanol UC rats, and FASL is the downstream target gene of miR-199a. TWP can reduce the UC's inflammatory effectively and decrease the up-regulated miR-199a in UC. FASL is up-regulated in UC's inflammatory activity. TWP can reduce downstream target gene FASL of miR-199a.