Chaperone-mediated autophagy (CMA) is a highly selective form of autophagy responsible for the degradation of specific cytosolic proteins within lysosomes. Recent research has established a significant correlation between CMA and colorectal cancer (CRC). However, the majority of current research focuses on tumor parenchymal cells, with limited attention paid to the expression and role of CMA in tumor stromal cells, particularly in tumor-associated macrophages (TAMs). In this study, we generated myeloid-specific LAMP2A-knockout and knock-in mice to investigate the role of macrophage CMA in dextran sodium sulfate (DSS)-induced colitis and azoxymethane/dextran sodium sulfate-induced CRC. Our findings indicated that the expression of LAMP2A, the rate-limiting component of CMA, was reduced in tumor-associated macrophages of both human and mouse CRC tissues. The knockout of LAMP2A in macrophages exacerbated experimentally induced colitis and colitis-related CRC, whereas its overexpression in macrophages alleviated the progression of colitis and CRC in mice. Notably, we observed increased angiogenesis within the tumor mass of CRC tissues from LAMP2A-mØKO mice. Mechanistically, LAMP2A deficiency elevated the protein levels of HIF-1α, thereby enhancing the secretion of its target genes, vascular endothelial growth factor A and IL-1β, which are 2 important proangiogenic cytokines. Our study suggests that the activation of CMA in macrophages may represent a promising therapeutic strategy for the treatment of CRC.

The use of hypoxia-preconditioned exosomes (HP-Exos) to modulate intestinal immunity in ulcerative colitis (UC) represents a promising therapeutic strategy. However, the effects of hypoxic preconditioning on exosomes derived from bone marrow mesenchymal stem cells (BMSCs) and the underlying mechanisms in UC treatment remain inadequately understood. This study sought to elucidate the regulatory roles and molecular mechanisms of HP-Exos in the context of UC. HP-Exos were isolated from BMSCs and characterized. We employed hypoxia-inducible factor 1α (HIF-1α)-silenced lentivirus-interfered HP-Exos to assess their effects in both in vivo and in vitro models. A series of experiments were conducted to evaluate the effects of HP-Exos on mitophagy, oxidative stress, and apoptosis in HT-29 cells and colonic tissues and to comprehensively analyze the immunoprotective mechanisms of HP-Exos. The results demonstrated that HP-Exos enhanced mitophagy, inhibited reactive oxygen species (ROS) accumulation and apoptosis in HT-29 cells and colon tissues, and upregulated the expression of Bcl-2 19-kDa interacting protein 3 (BNIP3), a downstream effector of HIF-1α. Conversely, HIF-1α knockdown markedly reversed the increase in mitophagy and the inhibition of apoptosis. Our findings indicate that HP-Exos protect against DSS-induced colitis by mitigating apoptosis and ROS production through HIF-1α-BNIP3-mediated mitophagy.

INTRODUCTION:Ulcerative colitis (UC) is a refractory disease with complex pathogenesis, and its pathogenesis is not clear. The present study aimed to investigate the potential target and related mechanism of Compound Sophora Decoction (CSD) in treating UC. METHODS:A network pharmacology approach predicted the components and targets of CSD to treat UC, and cell and animal experiments confirmed the findings of the approach and a new target for CSD treatment of UC. RESULTS:A total of 155 potential targets were identified for CSD treatment of UC, with some related to macrophage polarization, such as nitric oxide synthase (NOS2), also known as inducible nitric oxide synthase (iNOS). GO and KEGG enrichment analysis indicated that oxidative stress response and multiple inflammatory signaling pathways such as TNF-α may play a significant role. In vitro experiments revealed that Interferon-stimulated DNA (ISD) interference can cause polarization imbalances in Raw 264.7 and bone marrow-derived macrophages (BMDMs). Flow cytometry demonstrated that polarization of macrophages in the intestine, spleen, and lymph nodes in vivo was also unbalanced after dextran sulfate sodium (DSS) modeling with pathological intestinal injury. Both in vitro and in vivo studies indicated that after inducing inflammation, the levels of macrophage polarization-related markers (iNOS and Arg1) and inflammation-related factors (CCL17, IL10, TNF-α, and CXCL10) changed, accompanied by increased expression of cGAS. However, CSD treatment based on inflammation can inhibit the expression of cGAS protein and mRNA, lower the level of inflammatory factors, promote the expression of anti-inflammatory factors, and regulate macrophage polarization. CONCLUSION:We concluded that CSD alleviated DSS-induced UC by inhibiting cGAS, thus regulating macrophage polarization.

Macrophage-driven immune dysfunction of the intestinal mucosa is involved in the pathophysiology of ulcerative colitis (UC). Emerging evidence indicates that there is an elevation in miR-31-5p levels in UC, which is accompanied by a downregulation of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) expression. Nevertheless, the precise influence of miR-31-5p on macrophage polarization and the integrity of the intestinal epithelial barrier in UC remains to be fully elucidated. This study explored the role of miR-31-5p and AMPK in UC through a bioinformatics investigation. It investigated the potential of miR-31-5p antagomir to shift macrophages from pro-inflammatory M1 phenotype to anti-inflammatory M2 phenotype and enhance the intestinal mucosal barrier in DSS-induced UC mice. Additionally, RAW264.7 cells stimulated with LPS were employed to confirm the reversal of miR-31-5p antagomir's therapeutic effect under AMPK inhibition. The findings demonstrated that miR-31-5p antagomir penetrated colonic tissues and ameliorated DSS-induced experimental colitis. Transformation of spleen and mesenteric lymph node macrophages from M1 to M2 type was seen in the DSS+miR-31-5p antagomir group. AMPK/Sirt1 expression increased while NLRP3 expression decreased. Expression of M2-related genes and proteins was enhanced and that of the M1 phenotype suppressed. Tight junction proteins, ZO-1 and occludin, were increased. The therapeutic effects of miR-31-5p antagomir transfection into RAW264.7 cells were repressed when AMPK expression was inhibited. Therefore, our results suggest that suppression of miR-31-5p expression transformed macrophages from M1 to M2, ameliorated inflammation and repaired the intestinal epithelium to alleviate DSS-induced colitis. AMPK/Sirt1/NLRP3 was involved.

Macrophage polarization is closely associated with the pathogenesis of ulcerative colitis (UC). Quercetin, a flavonoid, has shown promise as a treatment for inflammatory diseases, but its specific mechanism of action remains unclear. This study investigates whether quercetin can regulate intestinal macrophage polarization and promote intestinal tissue repair via the cGAS-STING pathway for the treatment of UC. In vivo, mice with 3% DSS-induced UC were intraperitoneally injected with quercetin and RU.521 for 7 days, following which their general conditions and corresponding therapeutic effects were assessed. The impact of interferon-stimulated DNA (ISD) and quercetin on macrophage polarization and the cGAS-STING pathway was investigated using RAW264.7 cells and bone marrow-derived macrophages (BMDMs) in vitro. The results demonstrated that ISD induced M1 macrophage polarization and activated the cGAS-STING pathway in vitro, while quercetin reversed ISD’s inflammatory effects. In vivo, quercetin suppressed the cGAS-STING pathway in the intestinal macrophages of DSS-induced UC mice, which reduced M1 macrophage polarization, increased M2 polarization, and facilitated intestinal barrier repair in UC. Taken together, these findings provide new insights into the mechanisms via which quercetin could be used to treat UC.

AimsGastrointestinal (GI) dysfunction, as a common peripheral-organ complication after traumatic brain injury (TBI), is primarily characterized by gut inflammation and damage to the intestinal mucosal barrier (IMB). Previous studies have confirmed that TongQiao HuoXue Decoction (TQHXD) has strong anti-inflammatory properties and protects against gut injury. However, few have reported on the therapeutic effects of TQHXD in a TBI-induced GI dysfunction model. We aimed to explore the effects of TQHXD on TBI-induced GI dysfunction and the underlying mechanism thereof. MethodsWe assessed the protective effects and possible mechanism of TQHXD in treating TBI-induced GI dysfunction via gene engineering, histological staining, immunofluorescence (IF), 16S ribosomal ribonucleic acid (rRNA) sequencing, real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), Western blot (WB), and flow cytometry (FCM). ResultsTQHXD administration ameliorated TBI-induced GI dysfunction by modulating the abundance and structure of bacteria; reconstructing the destroyed epithelial and chemical barriers of the IMB; and improving M1/M2 macrophage, T-regulatory cell (Treg)/T helper 1 cell (Th-1), as well as Th-17/Treg ratios to preserve homeostasis of the intestinal immune barrier. Notably, Cluster of Differentiation 36 (CD36)/15-lipoxygenase (15-LO)/nuclear receptor subfamily 4 group A member 1 (NR4A1) signaling was markedly stimulated in colonic tissue of TQHXD-treated mice. However, insufficiency of both CD36 and (C-X3-C motif) chemokine receptor 1 (CX3CR1) worsened GI dysfunction induced by TBI, which could not be rescued by TQHXD. ConclusionTQHXD exerted therapeutic effects on TBI-induced GI dysfunction by regulating the intestinal biological, chemical, epithelial, and immune barriers of the IMB, and this effect resulted from the stimulation of CD36/NR4A1/15-LO signaling; however, it could not do so when CX3CR1 and CD36 were deficient. TQHXD might therefore be a potential drug candidate for treating TBI-induced GI dysfunction.

BackgroundDisruption of intestinal barrier function and an imbalance in intestinal immunity are crucial for the occurrence and development of ulcerative colitis. Because of their important roles in regulating inflammation and immunity, exosomes (Exos) released from bone marrow mesenchymal stem cells (BMSCs) may be useful for treating ulcerative colitis. The EphB/EphrinB signaling pathway plays a crucial role in the inflammatory process and the development and function of immune cells, and can mediate long-distance intercellular communication through extracellular vesicles. This study was conducted to explore the effects of pre-modified BMSC-Exos expressing EphB2 (EphB2-Exos) on immunoregulation in vitro.MethodsWe transfected a lentivirus vector encoding EphB2 into BMSCs and isolated EphB2-Exos from the culture supernatant. Inflammation and oxidative damage in the human colon adenocarcinoma cell line (Caco-2) were induced by dextran sulfate sodium/hydrogen peroxide. In addition, spleen CD4(+) T lymphocytes of rats were sorted in vitro. We conducted a series of experiments to explore the biological functions of EphB2-Exos.ResultsEphB2-Exos were successfully isolated and were found to significantly protect the activity, proliferation, and migration of Caco-2 cells that were inhibited by dextran sulfate sodium. EphB2-Exos alleviated inflammation and apoptosis and increased the activity of antioxidant enzymes while inhibiting oxidative stress in Caco-2 cells. EphB2-Exos restored intestinal barrier function by inhibiting the RhoA/ROCK pathway and regulated the polarization of CD4(+)T cells.ConclusionEphB2-Exos enhanced intestinal barrier function and regulated the immune balance by inhibiting the RhoA/ROCK pathway in vitro. These findings suggest that EphB2-Exos can be applied as a cell-free therapy for ulcerative colitis.

Traumatic brain injury (TBI) is the leading cause of disability and death, and the social burden of mortality and morbidity caused by TBI is significant. Under the influence of comprehensive factors, such as social environment, lifestyle, and employment type, the incidence of TBI continues to increase annually. Current pharmacotherapy of TBI mainly focuses on symptomatic supportive treatment, aiming to reduce intracranial pressure, ease pain, alleviate irritability, and fight infection. In this study, we summarized numerous studies covering the use of neuroprotective agents in different animal models and clinical trials after TBI. However, we found that no drug has been approved as specifically effective for the treatment of TBI. Effective therapeutic strategies for TBI remain an urgent need, and attention is turning toward traditional Chinese medicine. We analyzed the reasons why existing high-profile drugs had failed to show clinical benefits and offered our views on the research of traditional herbal medicine for treating TBI.

BACKGROUND:The mechanism of Heat Shock Protein 90 (HSP90) in Ulcerative Colitis (UC) has been studied, and mitogenic-activated protein kinases (MAPK) also contribute to the pathogenesis of UC. However, the effect of the HSP90/MAPK pathway in UC is still unclear. Therefore, the mainstay of this research is to explore the mechanism of action of this pathway in UC. Compound sophorae decoction (CSD), as a Chinese herbal decoction, can synergistically affect the above process.OBJECTIVE:This study aimed to uncover the synergistic effects of HSP90 inhibitors regulating the MAPK pathway for treating DSS-induced colitis in mice and the synergistic effects of CSD.METHODS:This experiment used oral administration of standard diets containing 3% dextran sodium sulfate (DSS) to establish an experimental colitis model in mice. The model was treated with HSP90 inhibitor, CSD, or dexamethasone. Mouse feces, mobility, body weight, colon length, and colon histopathology scores were recorded daily to assess the degree of colitis inflammation. Expression levels of HSP90 and MAPK pathway-related genes and proteins were evaluated by Western blot and qPCR. The evaluation of intestinal mucosal permeability was measured by enzyme-linked immunosorbent assay (ELISA), which could detect the protein level of D-Amino Acid Oxidase (DAO) and D-lactic acid (D-LA). The same went for downstream molecules AFT-2, p53, and apoptosis-related proteins BAX, BCL-2, Caspase3, and survivin in the MAPK pathway. Immunohistochemical measured p-38, p-JNK, and p-ERK expressions. JAM-A and claudin-1 connexin were tested by immunofluorescence staining. The TUNEL method was for measuring the apoptosis rate of colonic epithelial cells. CBA kit determined the level of inflammatory factors of colons.RESULTS:HSP90 inhibitor can improve the degree of pathological damage in the colon of mice treated with DSS, increase the mice's weight and the length of the colon, and significantly reduce the disease activity index (DAI) score. Intraperitoneal injection of HSP90 inhibitor can reduce the expression of MAPK pathway markers P38, JNK, ERK, and their phosphorylation and decrease the content of AFT-2 and p53, which is downstream of the MAPK pathway. In addition, treatment of the HSP90 inhibitor up-regulated the expression of anti-apoptotic proteins BCL-2 and survivin, as well as down-regulated apoptotic protein caspase3, BAX in the colon of mice with colitis. Lower levels of inflammatory factors such as IL-6, MCP-1, IFN-γ, TNF, IL-12p70, and increased IL-10 were observed after HSP90 inhibitor therapy. Furthermore, the combination treatment of CSD can enhance the effect of the single HSP90 inhibitor treatment and play a synergistic effect.CONCLUSION:These data suggest that an HSP90 inhibitor is available to treat UC by inhibiting the MAPK signaling pathway. This axis can restore the intestinal mucosa barrier's function by reducing intestinal mucosa's permeability and inhibiting apoptosis of intestinal epithelial cells. The specific mechanism is that HSP90 inhibitor can reduce the pathological damage and inflammation levels of colitis mice, and reduce the apoptosis rate of colonic epithelial cells and the mucosal permeability, thereby restoring the mucosal barrier function. During this process, CSD works synergistically to improve the therapeutic effect of the HSP90 inhibitor.

Objective Bone marrow-derived mesenchymal stem cells (BMSCs) are a kind of stem cells with high differentiation potential and immunomodulatory ability, which has a broad prospect in the treatment of inflammatory bowel disease. The aim of this study is to investigate whether BMSCs could improve TNBS-induced colitis in Sprague-Dawley (SD) rats through inducing Tregs differentiation by expressing PD-L1. Methods BMSCs were isolated and identified by flow cytometry before being transfected with PD-L1 siRNA recombinant lentiviral vector. Then SD rats were randomly divided into 4 groups. Colitis induced by TNBS (sigma Aldrich) except normal group. On the fourth day of modeling, the rats in BMSCs control group and PD-L1 siRNA BMSCs group were injected with corresponding BMSCs through tail vein for 1 week, the dose was 5 × 10 6 cells. The normal group and model group were given the same volume of PBS. Results PD-L1 siRNA BMSCs and BMSCs could reach colon tissue in TNBS-induced colitis. BMSCs control group significantly improved the clinical symptoms and histopathological severity of TNBS induced colitis, but PD-L1 siRNA BMSCs group did not. We found that the percentage of Tregs in spleen and mesenteric lymph nodes decreased, the expression of PD-L1, IL10, PTEN was down-regulated, and the expression of p-Akt and p-mTOR was up-regulated in colon tissue after PD-L1 siRNA intervention. Conclusion This study suggested that BMSCs can induce the differentiation of Treg through inhibiting Akt/mTOR pathway by expressing PD-L1, which can significantly improve the symptoms and pathological damage of ulcerative colitis rats and affect the immune function.

目的 研究复方苦参汤对实验性溃疡性结肠炎(UC)小鼠肠道T细胞分化的影响.方法 将32只健康雄性C57小鼠随机分为正常对照组、模型组、复方苦参汤组及美沙拉嗪组,每组8只,除正常对照组外,均予以3％ 葡聚糖硫酸钠饮用1周复制UC模型,复方苦参汤组及美沙拉嗪组从造模第1天开始予以相应药物灌胃处理,对照组及模型组予以等量磷酸盐缓冲液灌胃,连续给药7 d.第8天处死所有小鼠,评估结肠组织损伤情况;qRT-PCR及ELISA检测结肠组织IFNγ、IL-17A、TNF-α、RORγt表达情况;流式细胞术检测脾脏及肠系膜淋巴结中IFNγ及RORγt表达阳性的CD4+T细胞占比.结果 与对照组比较,模型组结肠黏膜损伤严重,疾病活动指数评分升高,结肠长度缩短,结肠组织中IFNγ、TNF-α、IL-17A、RORγt的mRNA及蛋白表达水平明显升高,脾脏及肠系膜淋巴结中IFNγ+及RORγt+的CD4+T细胞占比也显著上升;而与模型组比较,复方苦参汤组及美沙拉嗪组小鼠结肠组织损伤程度有所减轻,IFNγ、TNF-α、IL-17A、RO Rγt的表达均有不同程度降低,脾脏及肠系膜淋巴结T h1及T h17细胞占比降低.结论 复方苦参汤可通过降低T h1及T h17细胞分化及肠黏膜相关促炎因子表达缓解肠道炎症损伤.

Objective . Ulcerative colitis (UC) is closely related to immune response, in which Treg cells (Tregs) suppress the autoimmune response of effector T cells to maintain homeostasis. As a marker of endoplasmic reticulum stress (ERS), HSPA5 was highly expressed in the colon tissue of UC patients. This study is aimed at evaluating the therapeutic effect of HSPA5 inhibitor (HA15) on dextran sulfate sodium- (DSS-) induced ulcerative colitis in mice and explored the effect and related mechanism of HSPA5 inhibitor on the differentiation and function of Tregs. Methods . Thirty-two C57BL/6 mice were randomly divided into four groups (8 mice per group): normal control group, DSS model group, HSPA5 inhibitor (HA15) group (intraperitoneal injection), and dexamethasone (DXM) group (intraperitoneal injection). Except for the blank control group, the other groups were induced with 3% DSS for 7 days and then given corresponding intervention therapy for 7 days. Results . The disease activity index (DAI) score, colon length, histopathological changes, and scores of DSS-induced mice show that HA15 could significantly improve the degree of inflammation in ulcerative colitis. Moreover, HA15 can better inhibit the expression of HSPA5, HSPA1A, and CHIP in the colon and increase the level of FOXP3 mRNA. Finally, the content of Treg cells and the levels of IL-10 and TGF- β 1 were significantly increased, and the levels of IL-6 were significantly reduced. Conclusions . HA15 can improve the differentiation and function of Treg cells by inhibiting the HSPA1A/CHIP pathway, thereby improving ulcerative colitis. Therefore, inhibiting the expression of HSPA5 may serve as a new approach to treat ulcerative colitis.

Objective. Ulcerative colitis (UC) is closely related to immune response, in which Treg cells (Tregs) suppress the autoimmune response of effector T cells to maintain homeostasis. As a marker of endoplasmic reticulum stress (ERS), HSPA5 was highly expressed in the colon tissue of UC patients. This study is aimed at evaluating the therapeutic effect of HSPA5 inhibitor (HA15) on dextran sulfate sodium- (DSS-) induced ulcerative colitis in mice and explored the effect and related mechanism of HSPA5 inhibitor on the differentiation and function of Tregs. Methods. Thirty-two C57BL/6 mice were randomly divided into four groups (8 mice per group): normal control group, DSS model group, HSPA5 inhibitor (HA15) group (intraperitoneal injection), and dexamethasone (DXM) group (intraperitoneal injection). Except for the blank control group, the other groups were induced with 3% DSS for 7 days and then given corresponding intervention therapy for 7 days. Results. The disease activity index (DAI) score, colon length, histopathological changes, and scores of DSS-induced mice show that HA15 could significantly improve the degree of inflammation in ulcerative colitis. Moreover, HA15 can better inhibit the expression of HSPA5, HSPA1A, and CHIP in the colon and increase the level of FOXP3 mRNA. Finally, the content of Treg cells and the levels of IL-10 and TGF-β1 were significantly increased, and the levels of IL-6 were significantly reduced. Conclusions. HA15 can improve the differentiation and function of Treg cells by inhibiting the HSPA1A/CHIP pathway, thereby improving ulcerative colitis. Therefore, inhibiting the expression of HSPA5 may serve as a new approach to treat ulcerative colitis.

Traditional computed tomography (CT) contrast agents, such as iodine-containing small molecules (omnipaque), have limitations in some applications. The development of nanotechnology has made it possible to develop CT contrast agents based on this technology. In this study, a large number of surface functional groups of the fifth-generation polyamide-amine dendrimer (P5-NH2) were applied to functionally modify polyethylene glycol (PEG), targeting molecules, or drugs, which were used as the carrier of CT contrast agents. With the help of sodium borohydride (NaBH4), there was a rapid reduction. The fluorescein thiocyanate (FT) and PEG modified with lactobionic acid (PEG-LA) weres connected before gold coating to obtain gold nanoparticles coated with targeted dendrimer (Au(P5-LA)DENPs). In the experiment, the gold nanoparticles were characterized, and the liver cancer nude mouse model was established, so as to analyze the CT imaging performance of the material. Besides, the above was applied in the motor function of children with cerebral palsy, and the improvement effect of CT imaging combined with transcranial magnetic stimulation based on the preparation of nanomaterials on the movement function of children was analyzed and demonstrated with the help of graph theory. The results showed that the average particle size of gold nanoparticles was 1.88 nm. Within the range of 5 °C–50 °C and pH = 4–7, the physical properties of the aqueous solution of this material were stable. What’s more, the cell activity still exceeded 80% when the material concentration reached 2000 nm. The nude mouse model of liver cancer indicated that the CT imaging based on this material enhanced the image contrast effect of the tumor part, and the material had no obvious toxic and side effects. CT imaging based on the preparation of nanomaterials can promote transcranial magnetic stimulation to accelerate the efficiency of brain movement, accelerate the global and local information exchange and integration speed of brain network, thereby improving the movement function of children.

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.

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.