Effects of Echinacea purpurea Polysaccharide on IEC-6 Cell Proliferation

[Objective] This study was conducted to investigate the effects of Echi-nacea purpurea polysaccharides （EPS） on proliferation of rat intestinal epithelial cel IEC-6. [Method] The proliferation rate of IEC-6 cel s cultured in EPS at different concentrations and for different time was measured by MTT assay and analyzed by statistic methods. [Result] The proliferation rate of IEC-6 cel s cultured in EPS at al the concentrations and for different time was improved by different extents in com-parison with the control. In detail, 50 and 200 μg/ml EPS greatly improved the IEC-6 cel proliferation after 24 h of culture; then, the cel proliferation rate in the two treatments increased from 24 to 48 h, and declined from 48 to 72 h. The cel pro-liferation was also significantly improved by culturing in 100 μg/ml EPS for 72 h and in 500 μg/ml EPS for 48 h. After 48 h of culture, the proliferation rate of IEC-6 cel increased in a EPS dose-dependent manner. [Conclusion] EPS can promote IEC-6 cel proliferation, and thus improve the intestinal mucosal absorption and immune function of rat.

黑灵芝多糖对脂多糖诱导的IEC-6肠上皮细胞损伤的保护作用

目的:探究黑灵芝多糖(PSG-1)对脂多糖(LPS)诱导的IEC-6肠上皮细胞损伤的保护作用及其机制。方法:采用LPS构建肠上皮细胞IEC-6损伤模型,研究PSG-1对IEC-6细胞的干预效果。采用细胞计数盒(cck-8)法测定PSG-1干预对细胞活力的影响。运用western-blot技术探究细胞中肠道紧密连接蛋白和环氧化酶cox-2表达的变化,基于转录组测序技术分析PSG-1潜在的保护机制并对其进行验证。结果:PSG-1干预可以显著提升LPS造成的细胞活力降低和肠道紧密连接蛋白ZO-1、Claudin-1和Occludin的表达,而且PSG-1对LPS引起的cox-2异常高表达具有抑制效果。转录组测序及划痕试验和蛋白免疫印迹试验结果表明:PSG-1能显著增强细胞的迁移能力并抑制促凋亡蛋白Bax、Caspase-3和Caspase-9的表达。结论:PSG-1对LPS诱导的肠上皮细胞IEC-6具有显著的保护作用,细胞迁移和凋亡可能是PSG-1发挥其保护效应的关键途径。

Changes in Behavior and Amino Acid Neurotransmitters in the Brain of Rats with Seizure Induced by IL-1β or IL-6

To explore the mechanism of epilepsy induced by IL-1β and IL-6, the changes of glutamic acid (Glu) and GABA immunoreaction in the cerebral cortex and hippocampus of rats with seizure induced by IL-1β or IL-6 were studied. Rats were randomly divided into 3 groups: control group (intracerebroventricular injection (icv) of NS), IL-1β group (icv injection of IL-1β) and IL-6 group (i.c.v. injection of IL-6). 120 min after the icv injection of reagents of IL-1β or IL-6, behavioral changes were observed and Glu and GABA in the cerebral cortex and hippocampus were examined by means of immunohistochemistry. Our results showed that no seizure developed in the control group, while moderate seizure was observed in IL-1β group and IL-6 group. Compared with the controls, the immunoreaction of Glu was significantly increased, while GABA was obviously decreased in IL-1β group and IL-6 group after 120 min. Our study suggested that the IL-1β and IL-6 might promote and induce epilepsy by increasing Glu and decreasing GABA in the cerebral cortex and hippocampus.

The bio-active components of the Mongolian medicine Horcha-6 and therapeutic mechanism in the rat migraine model

Background:The active components of Horcha-6 were identified using liquid chromatography with tandem mass spectrometry.Also,we investigated the potential mechanisms that explain why Horcha-6 may be effective in treating migraines through the use of network pharmacology and a rat migraine model.Methods:After identifying the active components of Horcha-6,the corresponding genes of the active components’target were obtained from the Universal Protein database,and a“compound-target-disease”network was constructed using Cytoscape 3.9.0 software.For the in vivo experiments,nitroglycerin was injected intraperitoneally into rats to create a migraine model.Pre-treatment with Horcha-6 was administered orally for 14 days,and rats were subjected to migraine-related behavior tests.RNA sequencing was performed to identify the gene expression regulated by Horcha-6 in the trigeminal nerve.Results:A total of 903 chemical components of Horcha-6 have been collected in the liquid chromatography with tandem mass spectrometry.We discovered 55 of the Horcha-6 bio-active components that were evaluated based on their Percent Human Oral Absorption(≥30%)and DL values(≥0.185)on the traditional Chinese medicine systems pharmacology database.The“compound-target-disease”network contained 163 intersection targets with the migraine state.Gene Ontology analysis indicated that these components significantly regulated the immune response,vascular function,oxidative stress,etc.When Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed,we observed that most of the target genes were significantly enriched in the inflammation and neuro-related signaling pathway,toll-like receptor signaling pathway,neuroactive ligand-receptor interaction,etc.These predictions were further demonstrated via in vivo animal model experiments.The RNA sequencing results showed that 41 genes were down-regulated(P<0.05)and 86 genes were up-regulated(P<0.05)in the Horcha-6 treated group compared with the untreated group.Those genes were mainly involved in neuromodulation,vascular function,and hormone metabolism.Conclusion:The 55 bio-active components in Horcha-6 regulate inflammation,hormone metabolism,and neurotransmitters and have potential as a therapy to treat migraines.

The effects of aging on muscle loss and tissue-specific levels of NF-<i>κ</i>B and SIRT6 proteins in rats

The objective of this study was to examine the influence of aging on food intake, tissue and organ mass and NF-κB and SIRT6 levels in various tissues. The transcription factor, Nuclear Factor Kappa-B (NF-κB), is associated with both catabolic and anabolic pathways of muscle metabolism and may be involved in age-related muscle loss. SIRT6 is a member of the sirtuin family of proteins that function as protein lysine deacetylases and are associated with longevity in a number of organisms. Male Sprague-Dawley rats, aged 6 months (Adult) and 21 months (Old) were fed a commercially available diet for 10-17 days. Old rats consumed less food per body weight (BW) each day than Adult rats (1.45% g diet/g BW vs. 2.4% g diet/g BW). However, when intake data were expressed as g/diet per day there was no significant difference between groups. For skeletal muscle tissue, the average mass of gastrocnemius and soleus (g muscle/g BW) was significantly lower in Old rats. Levels of NF-κB (p65/RelA) and SIRT6 were measured by Western blot analysis in gastrocnemius, tibialis anterior, quadriceps, soleus, lung, heart, kidney and liver. NF-κB levels were higher in gastroc- nemius of Old rats compared to Adult rats. No significant age-specific differences in SIRT6 protein levels were noted in the tissues examined. Interestingly, when examined independent of age, levels of SIRT6 were significantly different between certain tissues. Data from this study suggest that aging affects muscle loss and NF-κB in a tissue-specific manner. Furthermore, these findings indicate tissue-specific but not age-specific differences in SIRT6 protein levels.

Effects of four traditional Chinese medicinal herbs on plasma level of IL-6 and induced TNF level in burned rats with wound infection

Objective : Effects of the decoction of Coptis chinensis Franch. , Scutellaria baicalensis Georgi , P. chinense schneid and Lonicera japonica Thunb. on burn rats with wound infection were observed. Methods : Fifty-six rats were randomly divided into three groups :sham burn , burn wound infected control(BWIC) and burn wound infected and treatment(BWIT). The plasma was harvested from the experimental rats. Plasma IL-6 activity was measured by using the IL-6-dependent cell line B9. The isolated mono-nuclear cells (MNCs) were induced by LPS and cultured for 24 h. TNF in the culture supernatant was detected by ELISA. Results: Plasma IL-6 levels of BWIC and BWIT were significantly elevated at postburn (PB) 24 and 48h , and IL-6 level of BWIC was significantly higher than that of BWIT at PB 48h (P<0. 01). The induced TNF level of BWIT was significantly higher than that of BWIC at PB 24 and 48 h , and that of sham burn at 24 h(P<0. 01). Conclusion : The results indicate that this decoction could inhibit MNCs releasing from pro-inflammatory cytokines which are believed as an endogenous pyrogen.

Differences in action potential propagation speed and axon initial segment plasticity between neurons from Sprague-Dawley rats and C57BL/6 mice

Action potentials(APs)in neurons are generated at the axon initial segment(AIS).AP dynamics,including initiation and propagation,are intimately associated with neuronal excitability and neurotransmitter release kinetics.Most learning and memory studies at the single-neuron level have relied on the use of animal models,most notably rodents.Here,we studied AP initiation and propagation in cultured hippocampal neurons from Sprague-Dawley(SD)rats and C57BL/6(C57)mice with genetically encoded voltage indicator(GEVI)-based voltage imaging.Our data showed that APs traveled bidirectionally in neurons from both species;forward-propagating APs(fpAPs)had a different speed than backpropagating APs(bpAPs).Additionally,we observed distinct AP propagation characteristics in AISs emerging from the somatic envelope compared to those originating from dendrites.Compared with rat neurons,mouse neurons exhibited higher bpAP speed and lower fpAP speed,more distally located ankyrin G(AnkG)in AISs,and longer Nav1.2 lengths in AISs.Moreover,during AIS plasticity,AnkG and Nav1.2 showed distal shifts in location and shorter lengths of labeled AISs in rat neurons;in mouse neurons,however,they showed a longer AnkG-labeled length and more distal Nav1.2 location.Our findings suggest that hippocampal neurons in SD rats and C57 mice may have different AP propagation speeds,different AnkG and Nav1.2 patterns in the AIS,and different AIS plasticity properties,indicating that comparisons between these species must be carefully considered.

Effect of Maxingganshi decoction on the expression of STAT4 and STAT6 in lung tissues of rats with chronic obstructive pulmonary disease

Objective:To investigate the effect of Maxingganshi decoction on the expression of STAT4 and STAT6 in lung tissues of rats with chronic obstructive pulmonary disease (COPD).Methods:A total of 30 COPD Wistar rat models with phlegm-heat depression and lung depression were established, with half male and half female. The 30 model rats were fed in different cages, weighed and labeled. They were randomly divided into three groups: Maxingganshi decoction group, roxithromycin tablets control group, model control group. At the same time, 10 normal rats were selected as a blank control group. The white blood cell count and other cell count levels in the alveolar lavage fluid of the rats in the four groups, as well as the STAT4 and STAT6 protein levels in the lung tissues, were observed and compared.Results: After treatment, the white cell counts in Maxingganshi decoction group were significantly higher than that of the model control group (P<0.05);lymph, neutral particles and eosinophil levels were significantly lower than those of the model control group (P<0.05). Compared with roxithromycin tablets control group, white blood cell count and other classification level of cell count in Maxingganshi decoction group were not significantly different (P>0.05). After treatment, STAT4 protein levels in roxithromycin tablets control group and Maxingganshi decoction group were lower than that in the model control group (P<0.05), and STAT6 protein levels in roxithromycin tablets control group and Maxingganshi decoction group were higher than that of the model control group (P<0.05), suggesting the two means of intervention in this study could inhabit the STAT4 protein expression in lung tissue of COPD rats and promote effect on STAT6 protein. In addition, the level of STAT4 and STAT6 in the Maxingganshi decoction group was not significantly different from that in the roxithromycin tablets control group (P>0.05), suggesting that the effect of Maxingganshi decoction was similar to that of roxithromycin tablets.Conclusions: The action mechanism of Maxingganshi decoction group treating COPD may be through the STAT4 and STAT6 protein expression level to impose an effect, and thus interfere with IL-12 / STAT4 and IL-4 / STAT6 these two signaling pathways of Th1 cells and Th2 cells in the body of the gene expression, inhibiting the Th1 polarization and adjusting the imbalance of Thl/Th2 cells, so as to lower inflammatory response mediated by T cells and various kinds of pathological damage.

Preparation of 6-[^(18)F]fluoro-L-DOPA and its biodistribution in normal and unilateral PD model rats

No-carrier-added 6-[^18F] fluoro-L-DOPA(6-FDOPA) was synthesized via a multistep procedure from a commercial available precursor,6-nitroveratraldehyde,The total synthesis time was 75min,with a radiochemical yield of (10±3)%,high radiochemical purity(>99%) and high enantiomeric purity(>95%).The biodistributions of 6-FDOPA in normal and unilateral PD model rats were measured.The results from normal rats showed the expected high concentration of radioactivity in striatum and low distrbutions in cerebrum,cortex and cerebellum.The ration of the radioactivity in striatum to cerebellum reached a peak value(5.9) at 60 min.In unilateral PD model rate.whose substania nigra of the right side had been damaged by pre-treated with 6-OHDA,the radioactive concentration in striatum of the damaged side was significantly lower than that of the undamaged side or that of both sides in striatum of control groups.

Feasibility of establishing model of Parkinson disease by injecting 6-hydroxydopamine at different parts of the nigrostriatal pathway in the brain of rats

BACKGROUND： Previous researches found that animal models with Parkinson disease （PD） could be established by injecting 6-hydroxydopamine （6-OHDA） into medial forebrain bundle （MFB）, substantia nigra compacta （SNC） and caudate-putamen complex （CPU） of the nigrostriatal pathway. OBJECTIVE ： To compare behavioral, biochemica 6-OHDA injections in the areas of MFB, SNC and DESIGN： Controlled observational study and histological properties of these rats undergoing the CPU respectively. SEI-IING： Department of Neurology, First Affiliated Hospital of Guangxi Medical University MATERIALS： A total of 64 adult female SD rats weighing 180-230 g were provided by the Animal Experimental Center of Guangxi Medical University. 6-OHDA （Sigma Company, USA）; Brain solid positioner （Standard model 51600, Stoelting Co., IL, USA）; rotational monitoring of little animal （type QL-1, USA）; high liquid chromatography （HLC, Waters Company）. METHOOS： The experiment was carried out in the Medical Experimental Center of Guangxi Medical University from February to December 2005. ① According to digital table, 64 SD rats were divided into MFB group, SNC group, CPU group and control group with 16 in each group. On the basis of the brain atlas of Paxinos, rats in the first three groups were injected with 5 μL 6-OHDA into right MFB （0 mm of line of incisor tooth, A/P 4.4 mm, L/R 1.2 mm, ON -7.8 mm）, SNC （line of incisor tooth just equal to horizon, A/P -4.8 mm, L/R 1.6 mm, ON -7.8 mm） and CPU （0 mm of line of incisor tooth, A/P 1.2 mm, L/R 2.7 mm, ON -5.4 mm）, respectively. The rats in control group were injected with 5 μL ascorbic acid solution （2 g/L）. One week after operation, 0.1 g/L apomorphine （Apo, 0.05 mg/kg） was subcutaneously injected into neck and then rotational behavior induced by Apo was recorded once a week for 8 weeks. The PD models were considered successful only when rotational times more than or equal to 7 times per minute. Eight weeks after operation, micro-perfusion was used to obtain micro-perfusate in bilateral CPU and contents of 3,4-dihydroxyphenylacetic acid （3,4-DOPAC） and homovanillic acid （HVA） were also measured. In addition, amount of tyrosine hydroxylase positive cells （TH＊） in SNC was counted with immuno- histochemical staining. MAIN OUTCOME MEASURES ： ① Successful rate of PD models; ② contents of dopamine and its metabolite in MFB, SNC and CPU groups and TH＊ amount. RESULTS： All 64 SD rats were involved in the final analysis. ③ Successful rate and rotational behavior： One week after operation, there were 6 successful models both in SNC and MFB groups; in the 2^nd week, there were 6 both in SNC and MFB groups and 1 in CPU group; in the 3^nd week, there were 1 in MFB group and 3 in CPU group; in the 4^nd week, there were 3 in CPU group. Otherwise, no successful case was found out in the next 3 weeks. Abnormal rotational behavior was not observed in control group. Four weeks after operation, successful rates were 81% （13/16） in MFB group, 75% （12/16） in SNC group and 44% （7/16） in CPU group.② Contents of 3, 4-DOPAC and HVA： Eight weeks after operation, contents in the SNC area of the injured side were lower than those on non-lesion side （P 〈 0.01）.③Changes of TH＋ amount： Eight weeks after operation, TH＋ amount in the SNC area of the lesion side was lower than that on non-lesion side （P 〈 0.01 ）. CONCLUSION： Injecting 6-OHDA into MFB, SNC and CPU can damage dopaminergic cells and establish successful PD models.

Expression and significance of IL-6 mRNA and its protein in liver after injury in rats

In order to study the effects of IL-6 on the development of liver injury the distribution and cell localization of IL-6 mRNA and its protein in the liver was observed in rats after they were inflicted with 20%TBSA full-thickness burns and/or intraperitoneal injection of endotoxin (lipopolysaccharide, LPS). 156 Wistar rats were randomized into combined burn-endotixin group (CBEG), simple burn injury group (SBG),simple endotoxin group (SEG ) and normal control group (NCG). The changes of serum IL-6 level, and theexpression and localization of IL-6 and IL-6 mRNA in the liver were determined quantitatively, with imrnunohistochemistry (IHC) and in situ hybridization (ISH). It was found that serum IL-6 level showed 2 peak inthe first half hour and the 6th to 12th h after injury in CBEG and one peak in the 3rd h in SBG and in the 12thto 24th h in SEG. IL-6 was mainly located in the sinusoid endothelial cells and Kupffer cells and IL-6 mRNAwas distributed mainly in Kupffer cells, polymorphonuclear cells and macrophages. In addition, the expression levels of IL-6 and IL-6 mRNA were in agreement with the pathological changes of liver injury.It is concluded that IL-6 is one of the key cytokines in the pathogenesis of liver damages after combinedinjury of burns and endotoxin.

白术糖复合物对IEC-6细胞分化及绒毛蛋白表达的影响

目的:观察白术糖复合物对小肠隐窝细胞(IEC-6)细胞分化及分化标志物之一绒毛蛋白(villin)表达的影响。方法:IEC-6细胞分为空白对照组、阳性对照药胃泌素组(250μg/L)、白术糖复合物组(62.5、125、250、500、1000 mg/L),经相应处理后。光镜下观察细胞大体形态;透射电镜下观察细胞超微结构;RT-PCR法检测villin mRNA表达;免疫荧光法检测villin蛋白表达。结果:①给药7 d,光镜下空白对照组细胞处于幼稚状态;胃泌素和白术糖复合物组细胞处于分化状态;②给药20 d,电镜下空白对照组胞核较大,常染色质比例较高,细胞边缘仅见少量微绒毛;胃泌素和白术糖复合物组胞核较小,常/异染色质比例协调,细胞边缘生成大量微绒毛;③给药6 h,空白对照组细胞villin mRNA表达非常少,胃泌素组和白术糖复合物组villin mRNA表达均明显高于空白对照组;④给药7 d,免疫荧光检测发现空白对照组细胞仅有很少villin蛋白表达;胃泌素组表达非常强烈;白术糖复合物组表达稍弱于胃泌素组,但有特异性绒毛蛋白聚集点。结论:白术糖复合物有通过上调IEC-6细胞绒毛蛋白表达及分布而促其分化的作用。

IEC-6细胞迁移药理实验模型建立的研究(英文)

建立适合肠上皮整复研究的IEC-6细胞迁移药理实验模型,并观察黄芪糖复合物对细胞迁移的影响。方法：设立不同的细胞接种密度、划痕后观察时间、辅助材料Matrigel浓度、血清饥饿处理法、细胞迁移抑制剂DF-MO（二氟甲基鸟氨酸）浓度等以考察模型的建立条件,并在已建立的模型上观察受试药的效果。结果：①细胞宜以4×105/mL接种6孔板;②宜划痕后24 h观察细胞迁移数;③5%Matrigel为适宜浓度;④血清饥饿可造成细胞迁移数明显减少,应使用含血清的培养液。⑤DFMO 2.5~5 mmol/L为抑制细胞迁移适宜浓度。⑥黄芪糖复合物及阳性药精脒能促进细胞迁移。结论：建立了适宜药理实验的IEC-6细胞迁移模型,在此模型上可反映受试药对细胞正常迁移及迁移抑制的药效作用。

白术、黄芪、党参促进IEC-6细胞损伤后的快速修复

目的研究白术、黄芪、党参提取物对体外胃肠黏膜上皮细胞损伤的快速修复作用。方法采用Tips划痕法建立小肠上皮(IEC-6)细胞迁移模型,首先考察不同的IEC-6细胞接种密度、划痕后修复时间、不同的血清浓度以建立稳定的迁移模型;接着采用表皮生长因子(EGF)及其阻断剂AG1478对迁移模型进行评价;最后考察三味中药对IEC-6细胞迁移的作用效果。结果 (1)细胞的最佳接种密度为1×105个/m L;(2)宜在划痕后8 h计算细胞迁移率;(3)1%血清浓度为最适宜浓度;(4)EGF明显促进了IEC-6细胞迁移,AG1478明显抑制了细胞迁移;(5)白术、黄芪、党参提取物均可促进IEC-6细胞迁移。结论益气健脾中药可促进胃肠黏膜损伤后的快速修复过程,其机制是否与影响EGFR信号通路有关有待进一步研究。

唐古特大黄多糖促进IEC-6细胞增殖、移行作用及其可能的机制研究

目的观察唐古特大黄多糖组分之一(RTP1)对大鼠小肠隐窝上皮细胞株(IEC-6)细胞增殖、移行及其对细胞内鸟氨酸脱羧酶(ODC)活性和ODC蛋白表达的影响,观察其对胃肠黏膜的修复作用并探讨可能的机制。方法采用MTT法和体外肠上皮细胞移行模型,观察不同剂量的RTP1(10、30、100mg·L-1)处理IEC-6细胞24h后,细胞增殖和移行情况;Western blot检测ODC蛋白表达;通过测定L-[1-14C]鸟氨酸释放14CO2的量来检测ODC活性变化。结果RTP1在10、30、100mg·L-1时均可明显促进IEC-6细胞的增殖和移行,可增加ODC蛋白表达和ODC活性,并具有剂量依赖性,与空白对照组相比差异具有显著性(P<0·05或0·01)。结论RTP1可促进肠上皮细胞的增殖和移行,提示RTP1可能对肠黏膜的损伤修复具有直接作用。其作用机制可能与鸟氨酸脱羧酶表达增加和(或)活性增大有关。

党参白术提取物分别和合用诱导IEC-6细胞增殖分化的作用

目的 观察党参提取物 (D h)和白术提取物 (B t)对小肠隐窝细胞 (IEC 6 )增殖、分化的作用及其二者配伍对IEC 6细胞增殖的影响。方法 IEC 6细胞培养 2 4h ,加入不同剂量的D h、B t及二者的配伍溶液 ,2 4h后用MTT法观察细胞增殖 ,显微镜观察细胞分化的形态特征。结果 D h在 2 5 0mg·L-1以下剂量时对细胞增殖无影响 ,剂量提高到 5 0 0mg·L-1和 10 0 0mg·L-1时 ,明显促进细胞的增殖。B t各剂量均无促进细胞增殖的作用 ,在 5 0 0mg·L-1和10 0 0mg·L-1剂量时 ,细胞增殖反见减弱。二药配伍后 ,促进细胞增殖的作用明显增强 ,具有一定的剂量依赖关系 ,其效果优于D h ;IEC 6细胞在B t作用下 ,分化程度增高 ,低倍镜下细胞呈典型的上皮细胞形态 ,细胞排列呈条索状 ,部分呈腺管状排列。高倍镜下可观察到隐约有细胞连接样结构 ,上皮细胞似有微绒毛形成的趋势。D h处理的细胞分化程度较低 ,镜下所见多为未分化的梭状细胞 ,且排列不规则 ,无任何形成腺管的趋势和有细胞连接样结构。结论 D h能促进细胞增殖 ,B t可促进细胞分化 ,二者配伍后促进细胞增殖作用明显增强 。

不同伤情血清可有效激活IEC-6细胞PI3K/Akt通路

目的检测不同伤情血清对肠上皮细胞PI3K/Akt通路的激活。方法将对数生长期的IEC-6细胞无血清培养基培养24 h后,加入单放、单烧和复合伤24 h大鼠血清,并设正常大鼠血清组和无血清组作对照,刺激12 h后检测细胞AKT磷酸化水平。利用SELD I(surfaced enhanced laser desorption/ion ization)蛋白质芯片技术对单放、单烧、复合伤24 h大鼠血清差异蛋白进行筛选和分析。结果单放、单烧和复合伤24 h大鼠血清刺激后的IEC-6细胞Akt磷酸化水平较正常大鼠血清刺激的高,其中烧伤组最高。单放组血清与单烧血清比较有11个差异峰,复合伤组与单烧组相比有6个差异峰。结论单放、单烧和复合伤血清均可有效激活IEC-6细胞的PI3K/Akt通路,烧伤组最强,可能和烧伤组血清特殊变化有关。

白术提取物对IEC-6细胞迁移过程Rho GTPaes及肌球蛋白II表达的影响

目的观察白术提取物对小肠上皮细胞(IEC-6)迁移多胺介导钾通道激活信号通路指标Rho GTPaes(Rho A、Rac1和Cdc42)表达的影响,对肌球蛋白II(myosinⅡ)分布和表达的影响,以探讨益气健脾中药白术对胃肠黏膜损伤修复的作用机制。方法荧光定量PCR法检测Rho A、Rac1和Cdc42 mRNA表达;Western blot法检测Rho A、Rac1和Cdc42蛋白表达;免疫荧光法检测myosinⅡ蛋白分布和表达。结果白术提取物能提高细胞迁移过程Rho A、Rac1、Cdc42 mRNA和蛋白表达,逆转多胺合成抑制剂二氟甲基鸟氨酸(DFMO)所致的Rho A、Rac1、Cdc42 mRNA和蛋白表达抑制;提高细胞迁移过程myosinⅡ表达,从而增加细胞骨架应力纤维形成,改善DFMO所致的myosinⅡ蛋白表达和分布的抑制。结论白术提取物促进IEC-6细胞迁移的作用机制,与其影响多胺介导钾通道激活信号通路有关。