OBJECTIVE:To observe the effects of the Chinese medicine prescription Xiao-Cheng-Qi decoction (XCQD) on acute brain edema and inflammatory factors in rats with severe traumatic brain injury (sTBI).METHODS:A total of 108 male Sprague-Dawley (SD) rats were divided into control group, sham operation group, sTBI model group, and XCQD low, medium, high dose groups by random number table method, with 18 rats in each group. sTBI rat model was prepared according to the modified Freeney method. At 6 hours after injury, the XCQD low, medium, and high dose groups were given XCQD 1.80, 2.78, and 4.59 g/kg by gavage, respectively, and the other three groups were given the same amount of normal saline, once a day for 3 days. After 3 days of injury, rats in each group were sacrificed after the modified neurologic severity score (mNSS) assessed. Pathological changes of brain tissue were observed under light microscope after hematoxylin eosin (HE) staining, water content of brain tissue was measured by dry-wet specific gravity method, and the expressions of aquaporin 4 (AQP4), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue were detected by Western blotting. Serum TNF-α and IL-1β levels were detected by enzyme linked immunosorbent assay (ELISA).RESULTS:Compared with the normal group, the mNSS score of rats increased significantly, the structure of brain tissue was disordered, and pathological changes appeared such as inflammation, edema, pyknosis of nerve nuclei, water content, the protein expressions of AQP4, TNF-α and IL-1β in brain tissue, and the contents of TNF-α, IL-1β in serum were significantly increased. After XCQD intervention, the above indexes were significantly improved. Compared with sTBI model group, the mNSS score of XCQD medium and high dose groups significantly decreased (6.94±1.16, 6.88±1.02 vs. 8.61±1.09, both P < 0.05), and the pathological changes such as brain edema and inflammation were alleviated. Brain tissue water content, AQP4 protein expression and contents of serum TNF-α, IL-1β in XCQD low, medium, and high dose groups significantly decreased compared with sTBI model group [brain tissue water content: (78.25±0.71)%, (77.62±0.44)%, (76.70±0.74)% vs. (80.08±0.66)%; the expression of brain AQP4 protein (AQP4/β-actin): 0.86±0.13, 0.84±0.22, 0.65±0.13 vs. 1.08±0.14; serum TNF-α (ng/L): 106.34±15.07, 95.75±17.26, 89.00±17.36 vs. 141.96±29.47; serum IL-1β (ng/L): 90.41±12.88, 72.82±13.51, 71.32±16.79 vs. 128.57±22.56, respectively, all P < 0.05]. The protein expressions of TNF-α,IL-1β in brain tissue of XCQD medium and high dose groups also significantly decreased compared with sTBI model group [TNF-α (TNF-α/β-actin): 0.90±0.24, 0.79±0.35 vs. 1.17±0.15; IL-1β (IL-1β/β-actin): 0.91±0.21, 0.68±0.28 vs. 1.23±0.08, respectively, all P < 0.05]. Brain tissue water content, the expression of brain AQP4 protein, the levels of brain tissue and serum IL-1β in XCQD high dose group improved more significant than those of XCQD low dose group.CONCLUSIONS:XCQD can alleviate the acute brain edema in sTBI rats, and it is dose-dependent. The mechanism may be relevant to reduce the secondary inflammatory response of sTBI by inhibiting the expression of inflammatory factors TNF-α and IL-1β.

Objective: To study the effects of CX3CR1 on white matter injury, neurofunction, recognition, and expression of the CD36/15LO/NR4A1 signal in mice with traumatic brain injury (TBI). Methods: CX3CR1(GFP/GFP), CX3CR1(GFP/+) and C57BL/6 male mice were randomly divided into 3 groups. We used a controlled cortical impact (CCI) to establish a TBI model and T2wt MRI to detect the TBI lesion. FA and DTI allowed for quantitative evaluation of the structural integrity of white matter tracts. Several behavior tests were used to investigate nerve function; a computer-based tracing system was used to trace and analyze dendrites and cell bodies of microglia and astrocytes in the peri-lesional brain areas. We also used RT-PCR and western blot to detect the effect of CX3CL1/CX3CR1 axis on CD36/15LO/NR4A1 signal. Results: The fractional anisotropy (FA) at the corpus callosum area of brain was decreased at 3 days post TBI, the average lesion volume CX3CR1(GFP/GFP) group was increased, and the neurologic deficit scores of mice of Cx3Cr1(GFP/+) and wild-type groups were significantly increased compared to Cx3Cr1(GFP/GFP) group mice. In the Corner turn test, TBI induced impairments in forelimb function that were more severe than Cx3Cr11(GFP/+) and wild-type TBI mice. We operated the Y-maze at 3 days post-TBI and the NOR test at 28 days after TBI. There was a significant TBI effect induced in decreased percentage entries into the novel arm in Cx3Cr1(GFP/+) and wild-type TBI mice, compared with Cx3Cr1(GFP/GFP); Cx3Cr1(GFP/+). Wild-type mice showed decreased exploration time in new objects compared with Cx3Cr1(GFP/GFP). Those two behavior tests demonstrated that Cx3Cr1 knock-out increased the damage caused by TBI to memory. In the tail suspension and force swimming tests, there was no significant difference between those three groups. CD36 increased in Cx3Cr1(GFP/GFP) compared with the other three groups at 3 days after TBI. TBI inhibited the expression of NR4A1 at 3 d after damage. Cx3Cr1 deficiency can induce high expression of 15LO, this was unaffected by TBI. Conclusion: CX3CR1 deletion can enhance white matter injury. It increased the expression of CD36 and 15LO and increased expression of NR4A1. The lack of CX3CR1 can affect the recovery of nerve function. (C) 2021 Published by Elsevier Inc.

OBJECTIVE:To observe the dynamic of neurological severity scores (NSS) and the expressions of Wnt/β-catenin signaling pathway, brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in rats with severe traumatic brain injury (sTBI), and to explore the effect of Huoxue Huayu decoction.METHODS:A total of 126 Sprague-Dawley (SD) rats were randomly divided into seven groups by random number table with 18 rats in each group, namely control group (normal saline 2 kg/L), model group (normal saline 2 kg/L), brain protolysate group (BP group, 5.6 g/kg), Taohong Siwu decoction group (TH group, 10.2 g/kg), Xuefu Zhuyu decoction group (XF group, 15.6 g/kg), Tongqiao Huoxue decoction group (TQ group, 9.6 g/kg) and Buyang Huanwu decoction group (BY group, 28.7 g/kg). The sTBI rat model was reproduced by modified Feeney free fall method, and the rats in the control group were not treated with trauma. The rats in each group were intragastrical administered with corresponding drugs at 6 hours after injury, and the NSS scores were evaluated on the 1st, 3rd and 7th days after injury. After the hippocampus was harvested, the mRNA expressions of Wnt3a and β-catenin were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the positive expressions of BDNF and NGF were detected by immunohistochemistry.RESULTS:Compared with the control group, the rats in the model group showed obvious symptoms of craniocerebral injury at 1 day after injury, which was manifested as significantly increased NSS score, up-regulated mRNA expressions of Wnt3a and β-catenin, and increased positive expressions of BDNF and NGF, which indicated that the sTBI rat model was successfully prepared and presented a certain self-repair ability with the extension of time. Compared with the model group, NSS scores in the XF group, TQ group and BY group significantly decreased at 1 day after injury (6.6±1.5, 6.1±2.0, 5.7±2.4 vs. 9.4±1.5, all P < 0.05); however, the NSS scores in the BP group and TH group decreased significantly at 7 days after injury, and the NSS scores in the TQ group and BY group decreased more significantly than those in other drug groups. Compared with the model group, mRNA expressions of Wnt3a and β-catenin in the hippocampus of the BP group increased significantly at 1 day and 3 days after injury, respectively, and continued to increase with the extension of time. The mRNA expression levels of Wnt3a and β-catenin in the four groups of Huoxue Huayu decoction fluctuated to varying degrees from 1 day to 3 days after injury, but they were significantly higher than those in the model group at 7 days after injury, and the increase was more significant in the BY group [Wnt3a mRNA (2-ΔΔCt): 154.7±4.1 vs. 17.4±1.0, β-catenin mRNA (2-ΔΔCt): 17.05±0.45 vs. 2.74±0.13, both P < 0.05], and the second was the TQ group [Wnt3a mRNA (2-ΔΔCt): 126.6±2.8 vs. 17.4±1.0, β-catenin mRNA (2-ΔΔCt): 8.70±1.19 vs. 2.74±0.13, both P < 0.05]. Compared with the model group, the positive expressions of BDNF and NGF in the BP group increased significantly at 1 day after injury, but decreased after 3 days after peak. The positive expressions of BDNF and NGF in the four Huoxue Huayu decoction groups fluctuated to varying degrees from 1 day to 3 days after injury, but they were significantly higher than those in the model group at 7 days after injury, among which, the positive expressions of BDNF and NGF in the TQ group and BY group were significantly higher than those in the model group at 1 day after injury [BDNF positive cells (cells/MP): 56.4±6.2, 61.6±7.0 vs. 37.4±2.0, NGF positive cells (cells/MP): 58.4±5.0, 62.4±4.4 vs. 53.4±3.6, all P < 0.05], the increase amplitude at 7 days after injury was more significant than those in the other groups.CONCLUSIONS:Taohong Siwu decoction, Xuefu Zhuyu decoction, Tongqiao Huoxue decoction and Buyang Huanwu decoction have curative effect on the nerve regeneration and repair of rats with sTBI at acute stage, but the intensity of the effect is different. Buyang Huanwu decoction and Tongqiao Huoxue decoction have a fast and better effect.

With the rapid development of economy, transportation and industry, the incidence of severe traumatic brain injury (sTBI) is rising rapidly, which is one of the main traumatic diseases threatening human life. It is very difficult for sTBI patients to regenerate and repair the central nervous and recover the brain function. Moreover, no effective neuroprotective drug has been found in the treatment of sTBI patients. Seeking drugs to promote nerve repair has become a hot and difficult problem. It is widely accepted that thyroxine is one of the essential hormones in the human body, which not only promotes the growth and development of the nervous system, but also plays an important role in maintaining adult brain function. There are many reports of modern research on thyroxine, mainly focusing on the changes of thyroid hormone levels and their effects on the prognosis after injury. Besides, most of them are observed in clinical cases. Currently, there are few dynamic experimental studies about observing whether thyroxine can promote the repair of central nervous system at different stages after sTBI. In our previous experiment, we found that Wnt/β-catenin signaling pathway, whose functions are opposite to Notch signaling pathway, can be further activated by exogenous thyroxine in rats with sTBI. As a result, we are interested in the expression of Notch and Wnt/β-catenin signaling pathway in acute phase sTBI rats and the effect of thyroxine on those pathways. To investigate expression of Notch and Wnt/β-catenin signaling pathway in acute phase severe brain injury rats and the effect of thyroxine on those pathways by observing dynamically Notch and Wnt/β-catenin signaling pathway, NSS, GFAP, S100B, Bcl-2, Bax, etc. 108 rats were randomly divided into Group A (normal control group), Group B (normal-thyroxine group), Group C (TBI group), Group D (TBI+ low-dose thyroxine group), Group E (TBI + moderate-dose thyroxine) and Group F (TBI + high-dose thyroxine) with 18 rats in each group. The animal model was established according to Feeney's free-fall method, and administered with thyroxine or physiological saline at 6 h after sTBI. Six rats in each group were randomly killed on the 1st, 3rd and 7th days after intragastric administration. The changes of brain pathology and NSS were observed. The level of Wnt3a, β-catenin, Notch1 and Hes1 mRNA was detected by RT-PCR method, and the level of GFAP and S100B protein in serum was detected by ELISA. The expression of Bcl-2 and Bax was detected by immunohistochemistry. (1) There was no significant change in brain pathology and NSS in groups A and B, but the changes of brain pathology and NSS in group D, E and F were significantly less than those in group C, especially in groups E and F. (2) RT-PCR showed that there was no change in the expression of Wnt3a mRNA, β-catenin mRNA, Notch1 and Hes1 mRNA in groups A and B. Compared with group C, the expression of Wnt3a mRNA and β-catenin mRNA in group D increased significantly on the 7th day after sTBI, especially in groups E and F; expression of Notch1 and Hes1 mRNA in groups D, E and F increased gradually with time, especially in group F. (3) ELISA showed that Compared with group C, GFAP and S100B in group D did not change significantly at 3 time points, GFAP in groups E and F decreased gradually with time and reached the lowest value on the 7th day, and S100B in groups E and F decreased gradually with time, especially in group F. (4) Compared with group C, the expression of BCL-2 in brain tissue of groups D, E and F increased gradually with time, and peaked on the 7th day, and the increase of E and F was more obvious. The expression of Bax in brain tissue of group D, E and F decreased gradually with time. Exogenous thyroxine has no effect on Notch and Wnt/β-catenin signaling pathway in normal rats. After TBI, exogenous thyroxine can activate Notch and Wnt/β-catenin, and have a synergistic effect on the repair of central nervous system, which may be related to the up-regulation of Notch and Wnt/β-catenin signaling pathway mRNA expression and the increase of BDNF and NGF, and resist apoptosis in the brain of sTBI rats.

Colorectal cancer (CRC) is a common malignant tumor of the digestive system that can seriously threaten human health. Chinese matrine is known to have a wide range of antiviral and immunomodulatory effects. In this study, we evaluated the effect of matrine on ferroptosis using the HCT116 human colon cancer cell line. We evaluated cell viability and proliferation using the cell counting kit-8 assay and carried out cell clone formation experiment by measuring reactive oxygen species (ROS) production, as well as levels of glutathione (GSH), lipid peroxide (MDA), Fe2+, glutathione peroxidase 4 (GPX4), activating transcription factor 4 (ATF4), solute carrier family 7 member 11 (SLC7A11), transferrin receptor protein 2 (TFR2), and Sigma-1 receptor (Sigma-1R) in order to evaluate the redox status of cells. These results indicate that matrine can significantly reduce the cell viability of HCT116 and decrease cell proliferation. After treatment with matrine, ROS, Fe2+, and MDA levels increased significantly, while the GSH content decreased. In addition, the expression of GPX4, SLC7A11, and Sigma-1R decreased significantly, while the expression of ATF4 and TFR2 increased significantly. These results indicate that matrine can induce ferroptosis in CRC, which can provide new clues for further pharmacological research of matrine and provide experimental evidence for understanding its mechanism of inhibiting CRC.

OBJECTIVES:To investigate the effect of icariin (ICA) on early β-defensin-2 and T cell subsets in rats after tracheotomy. METHODS:A total of 54 SPF male Sprague-Dawley rats were randomly divided into a normal control group (group A), a model group (group B), and a model+ICA treatment group (group C), with 18 rats in each group. A tracheotomy intubation model of the B and C group was prepared. After 6 h of surgery, ICA intervention was given to group C. Groups A and B were given the same amount of normal saline. Lung tissue, alveolar lavage fluid and peripheral blood were taken at 24 h, 72 h and 168 h, respectively. The expression of rat β-defensin-2 mRNA in lung tissue was detected by RT-PCR. The content of β-defensin-2 in alveolar lavage fluid and peripheral blood serum was detected by ELISA. The content of peripheral blood T cell subsets (CD3+, CD4+, CD8+) was detected by flow cytometry, and the ratio of CD4+/CD8+ was calculated. RESULTS:After tracheotomy, the levels of β-defensin-2 mRNA and β-defensin-2 in lung tissue from the group B were increased significantly at 24 h, then they were decreased gradually, and decreased most significantly at 168 h (P<0.05). The content of β-defensin-2 in peripheral blood of group B decreased gradually, and the content of β-defensin-2 in 168 h was significantly lower than that in 24 h (P<0.05), but there was no significant difference between group B and group A (P>0.05). The level of CD3+ T cells in peripheral blood was significantly lower than that in the group A (P<0.05), but their was no significant difference in CD4+ and CD8+ T cells compared with group A (P>0.05). After ICA intervention in group C: lung tissue, alveolar lavage fluid, peripheral blood serum β-defensin-2 content, and peripheral blood CD3+ and CD4+ T cell levels were gradually increased, significantly higher than those in the group B (P<0.05). CD8+ T cell level was significantly lower than that in the group A at 24 h (P<0.05), the CD4+/CD8+ ratio was significantly higher at 168 h than those in the group A or B (both P<0.01). CONCLUSIONS:ICA can improve the early lung immune function in rats with tracheotomy, which might be related to up-regulation of β-defensin-2 in lung tissue and alveolar lavage fluid, concomitant with increases in CD3+ and CD4+ T cells and CD4+/CD8+ ratio in peripheral blood while reduction in CD8+ cells.

Epimedium, a traditional Chinese tonic herb, may improve the deficiency of kidney, strengthen the bones and muscles, and expel wind-damp. Icariin is one of the main flavonoids; Icariside Ⅱ (ICS-Ⅱ) is one of the main metabolites of icariin, with various pharmacological effects. This study briefly reviewed the recent advances in the bioavailability, anti-tumor, inhibition of osteoporosis and neuroprotective effects of ICS-Ⅱ, and provided a reference for further exploitation and application of ICS-Ⅱ.

目的 探讨不同活血化瘀方剂对重型颅脑损伤(sTBI)急性期大鼠的影响.方法 306只SD雄性大鼠随机分为正常组、模型组、脑蛋白水解物组、桃红四物汤、血府逐瘀汤组、通窍活血汤组、补阳还五汤组7组.除正常组外其余各组采用改良的Feeney'自由落体方法制备sTBI大鼠模型.造模成功后脑蛋白水解物组、桃红四物汤组、血府逐瘀汤组、通窍活血汤组、补阳还五汤组分别给予相应药物1.11 mg/200 g、2.04 g/200 g、3.12 g/200 g、1.92 g/200 g、5.74 g/200g灌胃,正常组和模型组给予生理盐水3 ml/200g灌胃,各组每日均灌胃1次,连续7天.灌胃后随时统计大鼠死亡率,在第1、3、7天分别观察大鼠脑组织病理评分,检测血清微管相关蛋白Tau、胶质纤维酸性蛋白(GFAP)含量.结果 实验期间正常组与脑蛋白水解物组大鼠无死亡,其余各组大鼠死亡主要集中在前4天,前4天与正常组比较,模型组、桃红四物汤组、血府逐瘀汤组大鼠死亡率都显著升高(P＜0.01);与模型组比较,桃红四物汤组、血府逐瘀汤组大鼠死亡率也明显升高(P＜0.01),而通窍活血汤组、补阳还五汤组差异无统计学意义(P＞0.05).与正常组同时间相比较,各给药组大鼠脑组织病理评分及血清Tau、GFAP水平在3个时间段均明显升高(P＜0.05).与模型组同时间比较,第1天血府逐瘀汤组、通窍活血汤组和补阳还五汤组大鼠病理评分升高,桃红四物汤组、血府逐瘀汤组、补阳还五汤血清GFAP蛋白水平升高(P＜0.05);而在第3天除桃红四物汤组外,其余各给药组病理评分较模型组降低;脑蛋白水解物组、通窍活血汤组、补阳还五汤血清Tau、GFAP水平降低(P＜0.05);第7天各给药组病理评分及血清Tau、GFAP水平均降低(P＜0.05). 结论 sTBI急性期使用桃红四物汤及血府逐瘀汤会加重病情,增加死亡风险.用活血化瘀药来治疗sTBI急性期的最佳时间段可能在发生损伤第4天后,并且以通窍活血汤和补阳还五汤效果最好.

目的:探讨淫羊藿苷(ICA)和淫羊藿次苷Ⅱ(ICS-Ⅱ)对气管切开插管留置大鼠免疫功能与炎性反应的影响.方法:将72只SPF级雄性SD大鼠随机分成4组,正常对照组(A)、模型对照组(B)及模型治疗组(C、D),每组18只.制备大鼠气管切开插管留置模型.模型成功6 h后给予C组ICS-Ⅱ4 mg/(kg·d)、D组ICA 30 mg/(kg·d)灌胃,A、B组灌胃等量0.9％ 生理盐水.分别在24、72、168 h取外周血及肺泡灌洗液.应用流式细胞仪检测外周血T细胞亚群(CD4+、CD8+、CD4+/CD8+)水平,酶联免疫吸附法(ELISA)检测肺泡灌洗液sIgA、IL-6及外周血血清IL-6含量.结果:气管切开大鼠分别用ICA和ICS-Ⅱ干预后:C组CD4+T细胞数量在168 h明显高于B组(P<0.05),D组在72 h、168 h明显高于B组(均P<0.05);D组在168 h明显高于C组(P<0.05).C组CD8+T细胞数量在72 h、168 h明显低于B组(均P<0.05);D组与B组相比无明显差异(P>0.05);D组在72 h明显高于C组(P<0.05).C组CD4+/CD8+与B组相比无明显差异(P>0.05);D组在168 h明显高于A、B、C 3组(均P<0.05).C、D两组外周血IL-6含量在3个时段均明显低于A、B组(均P<0.05);且D组在168 h明显低于C组(P<0.05).C、D两组肺泡灌洗液IL-6含量在72 h、168 h明显低于B组,且呈逐渐降低趋势(均P<0.05).C、D两组sIgA含量在72 h、168 h明显高于B组(均P<0.05);D组在24 h、168 h明显高于C组(均P<0.05).结论:ICA和ICS-Ⅱ能够提高肺泡灌洗液sIgA含量,抑制肺泡灌洗液及外周血IL-6表达和改善外周血T细胞亚群失衡,对气管切开大鼠起到减轻炎症及提高免疫的作用,且ICA效果优于ICS-Ⅱ,尤以168 h显著.

目的:探讨气管切开插管后大鼠肺部形态学和肥大细胞活性动态变化及其意义,阐明气管切开插管大鼠肺部炎症发生的机制.方法:48只SPF级健康雄性SD大鼠随机分为正常对照组和气管切开插管组,每组24只.制作大鼠气管切开插管留置模型,在造模成功后分别于24、48、72和168h4个时间点处死大鼠,取肺组织和肺泡灌洗液,光学显微镜下观察肺组织病理变化,透射电镜观察2组大鼠肥大细胞的超微结构,ELISA法测定2组大鼠肺泡灌洗液中组胺、肿瘤坏死因子α (TNF-α)和白细胞介素6(IL-6)水平,免疫组织化学法测定2组大鼠肺组织中类胰蛋白酶β表达水平.结果:光镜下气管切开插管组大鼠肺组织于各个时间点均有不同程度的炎症等改变,Smith评分均高于正常对照组(P＜0.05);气管切开插管组内不同时间点比较,随着时间的延长,大鼠肺组织炎症等变化不断加重,Smith评分升高(P＜0.05).气管切开插管组大鼠各时间点肥大细胞出现不同程度的脱颗粒,其中以24 h脱颗粒最明显.气管切开插管组大鼠肺泡灌洗液中组胺水平在24、72和168 h3个时间点高于正常对照组(P＜0.05),在48 h时2组组胺水平比较差异无统计学意义(P＞0.05).气管切开插管组大鼠肺泡灌洗液中TNF-α和IL-6水平在不同时间点均明显高于正常对照组(P＜0.05),在24 h时达到峰值,随后下降(P＜0.05).气管切开插管组大鼠肺组织中类胰蛋白酶β表达水平在不同时间点均明显高于正常对照组(P＜0.05),在24 h时达到最高峰(P＜0.05).结论:气管切开后大鼠肺部肥大细胞活性明显增强,肥大细胞活性增强可能是气管切开大鼠肺部炎症发生的主要机制之一.

Objective To investigate the effect of exogenous thyroxine on the regeneration and repair of central nervous system in rats with severe traumatic brain injury.Methods A total of 108 male SD rats were randomly divided into Group A (normal group),Group B (thyroxine group),Group C (TBI group),Group D [TBI + low dose of thyroxine (6.5 μg/kg) group],Group E [TBI + medium dose of thyroxine (10 μg/kg) group)] and Group F [TBI +high dose of thyroxine (15.4 μg/kg)] by random number table method,with 18 rats in each group.The animal model was established according to the Feeney method.Groups A and C were given saline and thyroxine 6 hours after TBI.At 1 day,3 days and 7 days after injury,six rats were selected from each group according to random number table method for brain tissue harvest.The pathological changes of brain tissues were observed,and the expressions of Wnt3a and β-catenin,brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detected by RT-PCR and immunohistochemistry respectively.Results (1) There were no pathological changes in brain tissue in Groups A and B.The pathological changes in brain tissue of TBI rats in Groups D,E and F were significantly mitigated than those in Group C,and the effect of Groups E and F were more obvious.(2) RT-PCR showed that the expressions of Wnt3a and β-catenin mRNA in hippocampus of Group D at 7 days after injury were significantly higher [(6.46±2.02) vs.(4.08 ±1.06);(13.53 ±1.64) vs.(2.11 ±0.63);P＜0.05);Wnt3a mRNA expression of Groups E and F were increased significantly at 3 days and 7 days after injury [(5.45 ± 1.54) vs.(2.78±1.04),(8.59±1.88) vs.(4.08 ±1.06),(5.40±1.38) vs.(2.78±1.04),(8.63±1.74) vs.(4.08± 1.06);P ＜ 0.05)].The expressions of β-catenin mRNA were increased significantly at 1 day,3 days,and 7 days after injury [(3.10 ± 0.59) vs.(1.45±0.28),(5.28±0.62) vs.(1.45±0.28),(4.12±0.43) vs.(1.64± 0.31),(8.78±0.92) vs.(1.64±0.31),(23.80±2.38)vs.(2.11±0.63),(25.94±3.79) vs.(2.11 ± 0.63);P ＜ 0.05)].The expressions of Wnt3a and β-catenin in hippocampus of Group B did not change significantly.(3) Immunohistochemical staining showed that the expression of BDNF and NGF in hippocampus of B did not change significantly after thyroxine treatment,but that of BDNF and NGF in Group D did not change significantly after treatment.The expressions of BDNF and NGF of Groups E and F were significandy higher than that in Group C and reached the peak at 3 days after injury [BDNF:(77.3 ± 5.4) vs.(55.0 ± 5.6),(78.0 ± 7.8) vs.(55.0±5.6);NGF:(83.7±5.8) vs.(62.2±10.3),(86.8±4.8) vs.(62.2 ±10.3);P＜0.05].Conclusions Exogenous thyroxine has no effect on the Wnt/β-catenin pathway in normal rats.But exogenous thyroxine can significantly promote the centralis regeneration and repair in TBI rats,which might be associated with its role in upregnlating the expression of mRNA and increasing the secretion of BDNF and NGF.

目的:探讨淫羊藿次苷Ⅱ(icariside Ⅱ,ICS-Ⅱ)对气管切开插管留置大鼠肺泡灌洗液分泌型免疫球蛋白A(secretory immunoglobulinA,sIgA)、肺组织β-防御素-2(rat Beta-defensin-2,rBD2)和外周血白细胞介素-6(interleukin-6,IL-6)含量的影响.方法:将54只SPF级雄性SD大鼠随机分成3组,正常对照组(A)、模型对照组(B)及用药组(C),每组18只.制备大鼠气管切开插管留置模型.模型成功6h后给予C组ICS-Ⅱ[30 mg· (kg·d)-1]灌胃,A、B组灌胃等量生理盐水.分别于24h、72 h、168 h 3个时间段取外周血、肺泡灌洗液及肺组织,采用酶联免疫吸附法(ELISA)检测肺泡灌洗液sIgA及外周血IL-6含量,PCR检测肺组织rBD2 mRNA相对表达量.结果:气管切开后肺泡灌洗液sIgA含量24h即明显升高(P＜0.01),但在72 h后又降至正常水平;用ICS-Ⅱ干预后肺泡灌洗液sIgA含量在3个时间段均明显升高(P＜0.05),以72 h升高最明显.气管切开插管后大鼠外周血清IL-6含量72 h明显降低(P＜0.01),在24h和168 h没有明显变化;用ICS-Ⅱ干预后,IL-6含量在3个时间段均明显下降(P＜0.05).气管切开插管后24 h rBD2表达明显增加(P＜0.01),随后72 h、168 h时间段逐渐降低;用ICS-Ⅱ干预后,rBD2表达量没有升高,反而明显下降(P＜0.05).结论:ICS-Ⅱ能够提高气管切开插管大鼠sIgA分泌,改善肺部局部免疫功能.

目的 探讨淫羊藿苷及其代谢产物宝藿苷I对气管切开模型大鼠肺组织病理及炎症的影响.方法将72只SPF级雄性SD大鼠随机分成四组,正常对照组(A)、模型非用药组(B)、模型+宝藿苷I组(C)、模型+淫羊藿苷组(D),每组18只.制备大鼠气管切开插管留置模型.模型成功6 h后给予C组宝藿苷I,D组淫羊藿苷干预,A、B组给与等量0.9％ 生理盐水.分别在24、72、168 h取肺组织及肺泡灌洗液.肺组织用苏木精-伊红染色(HE染色),光镜下观察分析肺组织的病理学改变;ELISA检测肺泡灌洗液白细胞介素6(IL-6),肿瘤坏死因子α(TNF-α)表达.结果1)肺组织病理结果显示:气管切开后B组大鼠肺损伤程度随时间延长逐渐加重,明显高于A组(P<0.05).C、D组肺损伤程度均明显轻于B组,且D组在24、168 h均明显轻于C组(P<0.05).D组在168 h病理评分明显降低(与24 h比较P<0.01).(2)肺泡灌洗液IL-6、TNF-α含量:气管切开插管后大鼠肺泡灌洗液IL-6和TNF-α含量在三个时段均明显高于A组(P<0.05).用宝藿苷I和淫羊藿苷干预后,C、D组IL-6、TNF-α含量在72、168 h明显低于B组(P<0.05);且C、D组组内IL-6和TNF-α含量在72、168 h明显低于24 h(P<0.05);D组与C组相比,IL-6含量无明显统计学差异(P>0.05).D组TNF-α在72 h、168 h均明显低于C(P<0.05).结论宝藿苷I和淫羊藿苷对气管切开术后早期肺部炎症有一定疗效,其机制与抑制肺泡灌洗液炎症因子IL-6、TNF-α的表达,减轻肺组织损伤有关,且淫羊藿苷效果更佳.

目的 探讨肉苁蓉多糖对气管切开插管留置大鼠肺部免疫功能的影响.方法 54只SD大鼠随机分为正常组、模型组、肉苁蓉多糖组.除正常组外,其他组建立气管切开插管留置大鼠模型.造模6小时后肉苁蓉多糖组用肉苁蓉多糖灌胃,正常组和模型组予等量生理盐水灌胃,均每日1次.造模后第24小时、72小时、168小时三个时段取材,并检测大鼠肺组织β-防御素-2(rBD2) mRNA表达、肺泡灌洗液分泌性免疫球蛋白A(sIgA)及外周血IL-2、IL-6的含量.结果 (1)与正常组比较,模型组大鼠rBD2 mRNA表达在气管切开插管留置第24小时明显升高,随后明显下降,第168小时降到最低点(P＜0.05).与模型组比较,肉苁蓉多糖组大鼠rBD2 mRNA表达在第24小时降低,第72小时、168小时明显升高(P＜0.05).(2)与正常组比较,模型组大鼠肺泡灌洗液sIgA含量在气管切开插管留置第24小时明显升高(P＜0.05),在第72小时开始下降,逐渐回落至正常组水平.与模型组比较,肉苁蓉多糖组大鼠sIgA含量在第72及168小时明显升高(P＜0.05).(3)与正常组比较,模型组大鼠IL-2含量在第72小时升高(P＜0.05),其余时间段无明显改变(P＞0.05).与模型组比较,肉苁蓉多糖组大鼠IL-2含量在第24小时即明显升高,第72小时继续升高,在第168小时达到高峰.(4)与正常组比较,模型组大鼠IL-6含量在第24小时出现明显升高(P＜0.05),在第72小时稍有下降,在第168小时无明显变化(P＞0.05).与模型组比较,肉苁蓉多糖用药大鼠外周血IL-6水平在第24小时即显著降低,随后逐渐降低,至第168小时达最低值(P＜0.05).结论 肉苁蓉多糖能够改善气管切开插管留置大鼠肺部免疫功能.

OBJECTIVE:To observe the effects of diammonium glycyrrhizinate (DG) on nerve regeneration repair in rats with severe traumatic brain injury (STBI) from the perspective of Wnt/β-catenin signaling pathway.METHODS:Seventy-two Sprague-Dawle (SD) male rats were randomly divided into normal group, STBI model group, ganglioside (GA) treatment group and DG treatment group. The STBI animal model was reproduced referring to modified Feeney free fall impact model. No injury was made in normal group. Six hours after modeling, monosialotetrahexosylganglioside sodium injection and DG injection were injected via tail vein of rats in GA treatment group and DG treatment group respectively, once a day for 7 days. Normal group and STBI model group were given the same amount of normal saline. Six rats in each group were sacrificed on the 1st, 3rd and 7th day after the challenge for neurological severity score (NSS), and then the blood of abdominal aorta was drawn and brain tissue was harvested. The contents of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in serum were detected by enzyme linked immunosorbent assay (ELISA). The pathological changes of sub-granular zone (SGZ) were observed under light microscope after hematoxylin eosin (HE) staining. Real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the mRNA expressions of Wnt3a, β-catenin, glycogen synthetase kinase-3β (GSK-3β) and Axin.RESULTS:(1) There was no neurological deficit in the normal group and NSS was 0. NSS score of rats increased significantly on the first day after modeling, and then decreased gradually over time. NSS of the rats treated with GA and DG were significantly lower than that of the STBI model rats (score: 7.33±2.07, 6.17±2.23 vs. 9.33±1.63, both P < 0.01). Though NSS gradually decreased over time, the differences were still statistically significant on the 7th day (score: 2.67±0.82, 1.00±0.00 vs. 6.17±2.23, both P < 0.01), and NSS of DG treatment group was significantly lower than that of GA treatment group. (2) In SGZ of rats, cells were arranged in a compact and orderly way in the normal group, but neurons and tissues were damaged and destroyed at different time points in the STBI model group. After either GA or DG treatment, the damage of nerve tissue was improved gradually over time, and the effect of DG was more obvious. (3) In the normal group, the mRNA expressions of Wnt3a and β-catenin were almost not expressed, the mRNA expressions of GSK-3β and Axin were higher, and the contents of BDNF and NGF in serum were less. On the 1st day after STBI, the mRNA expressions of Wnt3a and β-catenin in hippocampus, the contents of BDNF and NGF in serum were significantly increased, and the mRNA expressions of GSK-3β and Axin were significantly decreased. The mRNA expressions of Wnt3a and β-catenin in the hippocampus and the contents of BDNF and NGF in serum were significantly higher than those in the model group 1 day after GA or DG was added, the mRNA expressions of GSK-3β and Axin were significantly decreased, and the effect of DG was more significant than that of GA [Wnt3a mRNA (2-ΔΔCt): 3.51±0.14 vs. 2.93±0.05, β-catenin mRNA (2-ΔΔCt): 1.90±0.08 vs. 1.75±0.04, BDNF (ng/L): 4.06±0.55 vs. 3.16±0.64, NGF (ng/L): 9.53±1.08 vs. 7.26±0.43, GSK-3β mRNA (2-ΔΔCt): 0.75±0.01 vs. 0.79±0.01, Axin mRNA (2-ΔΔCt): 0.74±0.02 vs. 0.76±0.02, all P < 0.05]. It was gradually increasing or decreasing over time and the difference was still statistically significant up to the 7th day.CONCLUSIONS:DG can promote the recovery of nerve function in rats with STBI, and its mechanism may be related to the regeneration of nerve cells proliferation and differentiation by Wnt/β-catenin signaling pathway and the reconstruction of nerve tissue in SGZ of hippocampus.

Objective:To evaluate the effects of Epimedium polysaccharides (EPS) on pulmonary immune function in rats with tracheostomy.Methods:54 SD rats were randomly divided into three groups (n =18per group):a normal control group (group A),a model group (group B),and EPS group (group C).The rats in group B and group C were subjected to tracheotomy.After 6 h,rats in group C were treated with EPS via gavage,and those in groups A and B were treated with equal volume of saline.Secretory immunoglobulin A (sIgA) and pulmonary surfactant-related protein A (SP-A) levels in bronchoalveolar lavage fluid (BALF) and serum interleukin-2 (IL-2) level were detected by enzyme-linked immunosorbent assay (ELISA).The mRNA expression of β-defensin-2 (rBD2) mRNA in lung tissue was determined by real-time fluorescent quantitative PCR (qPCR).Results:(1) rBD-2 mRNA expression in group B was significantly higher than that in group A at 24 h after tracheostomy (P＜0.05).Compared with group B,the expression of rBD-2 mRNA in group C was decreased at 24 h,while increased significantly at 72 h and 168 h (P＜0.05).(2) The sIgA content in group B was significantly higher than that in group A after 24 h and 72 h of tracheotomy (P＜0.05),and then gradually decreased to a normal level (P＞0.05).Compared with group B,the content of sIgA in group C was notably elevated at 24 h and 168 h post-tracheotomy (P＜0.05).(3) The content of SP-A in group B was higher than that in group A at each time point,but was gradually decreased as intubation time prolonged (P＜0.05).The content of SP-A in group C was reduced at 24 h of modeling but increased with time prolonged,and was lower than that in group B at 168 h (P＜0.05).(4) The level of IL-2 in group B was higher than that in group A at 24 h post-tracheotomy (P＜0.05).Compared with group B,the serum IL-2 level in group C was markedly increased at 72 h and 168 h (P＜0.05),but there was no significant difference in IL-2 level between groups B and C at 24 h (P＞0.05).Conclusion:EPS could improve the pulmonary immune function of tracheostomy rats by regulating the rBD2 expression,elevate the contents of sIgA and SP-A in BALF and IL-2 level in serum.

Objective To explore the effect of Epimedium brevicornu Maxim. on immune function in rats with trachea cannula retention after tracheostomy.Methods Fifty-four SD rats were randomly divided into normal control group(group A),model group without medication(group B),and model group with medication(group C),with 18 rats in each group.Rat model with tracheostomy and trachea cannula retention was established in group B and group C.After 6 h of modeling,intragastric administration with water extract of Epimedium brevicornu Maxim.was performed in group C,and intragastric administration with same volume of normal saline was performed in the other two groups.At the 24th,72th and 168th hour after intragastric administration,β-defensin-2(BD-2) mRNA expression level in the lung tissues,interleukin 2(IL-2) level and T cell subsets counts in the peripheral blood were detected in 6 rats of each group.Results (1)In group B,the relative expression levels of BD-2 mRNA at the 72th and 168th hour were lower than the level at the 24th hour(P<0.05), and the level in group B increased at the 24th hour but decreased at the 168th hour compared to the level in group A(P<0.05).At the 24th,72th and 168th hour,the relative expression levels of BD-2 mRNA were higher in group C compared to the levels in group B(P<0.05). (2)At the 72th and 168th hour,the levels of IL-2 and CD4+were higher in group C compared to the levels in group B(all P<0.05).At the 24th hour,the CD3+level in group B was lower than that in group A,but the level in group C was higher than that in group B(all P<0.05). There were no significant differences in the CD8+level at the 24th or 72th hour among the three groups(P>0.05),but the level at the 168th hour was lower in group C compared to the level in group A(P <0.05).Conclusion Epimedium brevicornu Maxim. can improve the immune function in rats with trachea cannula retention after tracheostomy by up-regulating the expression of BD-2 mRNA in pulmonary tissues and the expressions of CD4+T cell and IL-2 in blood as well as down-regulating the expression of CD4+T cell.

目的:探讨淫羊藿苷对气管切开插管大鼠肺部肥大细胞活性及炎性反应的影响.方法:将SPF级健康雄性SD大鼠分成正常对照(A)组、气管切开插管留置模型非用药(B)组、气管切开插管淫羊藿苷用药(C)组.造模成功6h后C组予淫羊藿苷灌胃,A、B组予等量0.9％氯化钠溶液.于24、72、168h 3个时间段每组分别处死6只大鼠进行取材检测.结果:光镜下,B、C组肺组织病理各个时段均有不同程度的炎性反应等改变,Smith评分均高于A组(P＜0.05);但C组病变明显小于B组(72h后,P＜0.05).透射电镜下B组各时段内肥大细胞不同程度出现脱颗粒,24h时最明显,但C组程度明显低于B组(P＜0.05).B、C组β-类胰蛋白酶表达在3个时段均高于A组(P＜0.05);从72h后C组明显低于B组(P＜0.05),以168h为最明显.B、C组组胺含量在3个时间段高于A组(P＜0.05),B、C两组比较无差异.B、C两组TNF-α、IL-6含量变化基本一致,在不同时间段均明显高于A组(P＜0.05),24h时达到峰值,随后下降.结论:淫羊藿苷可减轻气管切开插管大鼠肺部炎性反应,其机制与抑制肥大细胞活性有关.

Objective:To investigate the effect and mechanisms of Xuefu Zhuyu decoction on severe traumatic brain injury (STBI) in rats.Methods:A total of 72 SD rats were randomly divided into group A (normal control group),group B (model group) and group C (Xuefu Zhuyu decoction group),with 24 cases in each group.Modified Feeney's method was used to establish a rat model of STBI.After 6 h of modeling,the rats in group C were given Xuefu Zhuyu decoction by gavage,and rats in group A and B were given equal volume of saline.Eight rats from each group were sacrificed at 24 h,72 h and 168 h post-modeling.The neurological deficit score (NSS) and the apoptosis rate of brain cells were detected.The protein expressions of Bcl-2 and Bax were determined by immunohistochemistry.Results:Compared with group A,the NSS score in group B was significantly increased (P＜0.05),and reached peak at 24 h,then gradually decreased and came to its lowest level at 168 h.However,the NSS score in group C was markedly decreased compared with group B (P＜0.05).The apoptosis rate of brain cells in group B was higher than that in group A,as evidenced by TUNEL staining (P＜0.05).It peaked at 72 h and then decreased.Compared with group B,the apoptosis rate of brain cells in group C was reduced significantly (P＜0.05).There was no obvious change in the apoptosis rate from 24 h to 72 h in group C (P＞0.05),but it reduced at 168 h (P＜0.05).Compared with group A,the expressions of Bcl-2 and Bax in group B were increased (P＜0.05).The Bax protein level in group B reached the peak at 72 h,and then reduced.Compared with group B,Bax protein level in group C was decreased notably at 72 h and 168 h (P＜0.05),while Bcl-2 was increased significantly (P＜0.05).The Bcl-2 expression in group C increased gradually and reached peak at 168 h,while Bax consistently decreased and descended to the lowest level at 168 h.Conclusion:Xuefu Zhuyu decoction could improve the neurological function in rats with STBI,and its mechanism might be related to the regulation of Bcl-2/Bax expression and the inhibition of brain cell apoptosis.