Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific...Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.展开更多
目的:探讨同型半胱氨酸(Hcy)促进单核细胞趋化蛋白-1(MCP-1)表达诱导巨噬细胞浸润加重急性缺血性卒中(AIS)的机制。方法:采用线栓法制备局灶性脑缺血再灌注小鼠模型。采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑缺血严重程度,HE染色...目的:探讨同型半胱氨酸(Hcy)促进单核细胞趋化蛋白-1(MCP-1)表达诱导巨噬细胞浸润加重急性缺血性卒中(AIS)的机制。方法:采用线栓法制备局灶性脑缺血再灌注小鼠模型。采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑缺血严重程度,HE染色法检测缺血脑组织神经细胞损伤状况和炎症细胞浸润程度;高效液相色谱法检测急性期小鼠脑组织Hcy表达;免疫组织化学染色、Western blot、real time RT-PCR法检测小鼠脑组织MCP-1表达。Transwell小室检测巨噬细胞迁移程度,TUNEL法检测Hcy对内皮细胞影响;利用MCP-1中和抗体观察对巨噬细胞迁移的影响。结果:与假手术组比较,模型组小鼠右侧大脑半球梗死明显,缺血脑组织损伤严重,神经细胞死亡并伴有大量炎症细胞浸润;Hcy水平显著升高,MCP-1蛋白和mRNA表达均明显增加。Hcy刺激小鼠脑血管内皮细胞后可诱导NF-κB(p65)入核,促进巨噬细胞迁移,促进内皮细胞凋亡,给予NF-κB(p65)抑制剂后MCP-1生成减少;MCP-1中和抗体明显抑制巨噬细胞迁移。结论:Hcy、MCP-1与脑缺血再灌注炎性反应相关,且Hcy可能通过NF-κB通路促进MCP-1表达,发挥诱导巨噬细胞浸润作用,加重脑缺血损伤。展开更多
目的探究人乳腺癌裸鼠移植动物模型体内孕激素受体膜组分1(PGRMC1)在黄体酮与炔诺酮(NET)刺激下对乳腺癌异种移植瘤增殖影响的差异性。方法构建PGRMC1稳定表达的人乳腺癌细胞系(MCF7),36只雌性BALB/c-nu裸鼠按随机数字表法分为MCF7/HA-P...目的探究人乳腺癌裸鼠移植动物模型体内孕激素受体膜组分1(PGRMC1)在黄体酮与炔诺酮(NET)刺激下对乳腺癌异种移植瘤增殖影响的差异性。方法构建PGRMC1稳定表达的人乳腺癌细胞系(MCF7),36只雌性BALB/c-nu裸鼠按随机数字表法分为MCF7/HA-PGRMC1组(n=18)和MCF7/HA-Vector组(n=18)。各鼠于颈部皮下包埋雌二醇(E_(2))缓释片。24 h后两组裸鼠左、右侧面皮下分别注射MCF7/HA-PGRMC1和MCF7/HA-Vector细胞悬液150μL,注射细胞数为1×107个。肿瘤生长至100 mm 3左右,将两组裸鼠进行组内分组,分别皮下联合包埋安慰剂、黄体酮缓释片、NET缓释片,即E_(2)+安慰剂亚组(n=6)、E_(2)+黄体酮亚组(n=6)、E_(2)+NET亚组(n=6)。记录肿瘤体积变化,绘制肿瘤生长曲线。45 d后完整摘除肿瘤组织免疫组化染色检测PGRMC1和Ki67表达情况。结果MCF7/HA-PGRMC1株系构建成功,在MCF7/HA-PGRMC1组中,与E_(2)+安慰剂亚组相比,E_(2)+黄体酮亚组肿瘤增长差异无统计学意义(P>0.05),而E_(2)+NET亚组肿瘤明显生长(P<0.05)。在MCF7/HA-Vector组中,与E_(2)+安慰剂亚组相比,E_(2)+NET亚组、E_(2)+黄体酮亚组的肿瘤增长差异均无统计学意义(P>0.05)。在MCF7/HA-PGRMC1组中,E_(2)+NET亚组Ki67呈强阳性表达,PGRMC1与Ki67表达呈正相关(r=0.892,P=0.014)。结论该实验在裸鼠体内证实PGRMC1过表达可特异性介导NET联合E_(2)的促乳腺癌增殖效应,而联合黄体酮并没有促进额外增殖。这一发现为临床激素治疗中孕激素类型的选择提供了关键的动物实验依据。展开更多
AIM: To characterize high-mobility group protein 1-toll-like receptor 4(HMGB1-TLR4) and downstream signaling pathways in intestinal ischemia/reperfusion(I/R) injury.METHODS: Forty specific-pathogen-free male C57BL/6 m...AIM: To characterize high-mobility group protein 1-toll-like receptor 4(HMGB1-TLR4) and downstream signaling pathways in intestinal ischemia/reperfusion(I/R) injury.METHODS: Forty specific-pathogen-free male C57BL/6 mice were randomly divided into five groups(n = 8 per group): sham, control, anti-HMGB1, anti-myeloid differentiation gene 88(My D88), and anti-translocatingchain-associating membrane protein(TRIF) antibody groups. Vehicle with the control Ig G antibody, antiHMGB1, anti-My D88, or anti-TRIF antibodies(all 1 mg/kg, 0.025%) were injected via the caudal vein 30 min prior to ischemia. After anesthetization, the abdominal wall was opened and the superior mesenteric artery was exposed, followed by 60 min mesenteric ischemia and then 60 min reperfusion. For the sham group, the abdominal wall was opened for 120 min without I/R. Levels of serum nuclear factor(NF)-κB p65, interleukin(IL)-6, and tumor necrosis factor(TNF)-α were measured, along with myeloperoxidase activity in the lung and liver. Inaddition,morphologic changes that occurred in the lung and intestinal tissues were evaluated. Levels of m RNA transcripts encoding HMGB1 and NF-κB were measured by real-time quantitative PCR, and levels of HMGB1 and NF-κB protein were measured by Western blot. Results were analyzed using one-way analysis of variance.RESULTS: Blocking HMGB 1, MyD 8 8, and TRIF expression by injecting anti-HMGB1, anti-My D88, or anti-TRIF antibodies prior to ischemia reduced the levels of inflammatory cytokines in serum; NF-κB p65: 104.64 ± 11.89, 228.53 ± 24.85, 145.00 ± 33.63, 191.12 ± 13.22, and 183.73 ± 10.81(P < 0.05); IL-6: 50.02 ± 6.33, 104.91 ± 31.18, 62.28 ± 6.73, 85.90 ± 17.37, and 78.14 ± 7.38(P < 0.05); TNF-α, 43.79 ± 4.18, 70.81 ± 6.97, 52.76 ± 5.71, 63.19 ± 5.47, and 59.70 ± 4.63(P < 0.05) for the sham, control, anti-HMGB1, anti-My D88, and anti-TRIF groups, respectively(all in pg/m L).Antibodies also alleviated tissue injury in the lung and small intestine compared with the control group in the mouse intestinal I/R model. The administration of antiHMGB1, anti-My D88, and anti-TRIF antibodies markedly reduced damage caused by I/R, for which anti-HMGB1 antibody had the most obvious effect.CONCLUSION: HMGB1 and its downstream signaling pathway play important roles in the mouse intestinal I/R injury, and the effect of the TRIF-dependent pathway is slightly greater.展开更多
The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardi...The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts(CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pc DNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·m L^(-1) of osthole for 24 h, the m RNA and protein expression levels of collagensⅠand III were reduced. The collagen Ⅰ/III ratio was also reduced. The m RNA and protein expression levels of TGF-β1, TβRⅠ, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen Ⅰ and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.展开更多
Objective： Recent studies have shown that the local expression of soluble interleukin （IL） -1 receptor type Ⅱ （slL-1 R Ⅱ ） in endometrial tissue of women with endometriosis is decreased, and the depression of I...Objective： Recent studies have shown that the local expression of soluble interleukin （IL） -1 receptor type Ⅱ （slL-1 R Ⅱ ） in endometrial tissue of women with endometriosis is decreased, and the depression of IL-1 R Ⅱ was more significant in infertile women than that in fertile women with endometriosis. In this research, we investigated the remedial effect of slL-1-R Ⅱ administration on endometriosis in the nude mouse model. Methods： Nineteen nude model mice with endometriosis were randomly divided into three groups： group A was treated by intraperitoneal administration with only slL-1 R Ⅱ for two weeks, group B was similarly treated with only IL- 1, and group C （control） was administered saline. After 2 weeks, the size of the ectopic endometrial lesions was calculated, and the expression of vascular endothelial growth factor （VEGF） and B-cell lymphoma leukemia-2 （Bcl- 2） were detected by immunohistochemistry. The IL-8 and VEGF levels in the peritoneal fluid （PF） and serum were also measured by enzyme-linked immunosorbent assay （ELISA）. Results： The mean size of ectopic endometrial lesion did not differ between the three groups （P 〉 0.05）. Compared with the control, the expression of VEGF and Bcl-2 was significantly lower in group A, and higher in group B. In the three groups, the levels of IL-8 in the PF and serum were highest in group A, and lowest in group B. Conclusion： slL-1 R Ⅱ may suppresse hyperplasia of ectopic endometriosis, perhaps by reducing the expression of certain cytokines, such as VEGF, IL-8, and Bcl-2, which could provide a new clinical strategy for the treatment of endometriosis.展开更多
This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybr...This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups： control group, D2 group （2 days after pregnancy）, D4 group （4 days after pregnancy）, D6 group （6 days after pregnancy） and D8 group （8 days after pregnancy）, each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups （P〈0.01）. Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy （day 2）, and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice （P〈0.01）. In situ hybridization showed Ang-I mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 （P〈0.01）. It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.展开更多
Background The induced pluripotent stem cell （iPSC） has shown great potential in cellular therapy of myocardial infarction （MI）, while its application is hampered by the low efficiency of cardiomyocyte differentia...Background The induced pluripotent stem cell （iPSC） has shown great potential in cellular therapy of myocardial infarction （MI）, while its application is hampered by the low efficiency of cardiomyocyte differentiation. The present study was designed to investigate the effects of cardiotrophin-1 （CT-1） on cardiomyocyte differentiation from mouse induced pluripotent stem cells （miPSCs） and the underlying mechanisms involved. Methods The optimal treatment condition for cardiomyocyte differentiation from miPSCs was established with ideal concentration （10 ng/mL） and duration （from day 3 to day 14） of CT-1 administration. Up-regulated expression of cardiac specific genes that accounted for embryonic cardiogenesis was observed by quantitative RT-PCR. Elevated amount of a-myosin heavy chain （ct-MHC） and cardiac troponin I （cTn I） positive cells were detected by immunofluorescence staining and flow cytometry analysis in CT- 1 group. Results Transmission electron microscopic analysis revealed that cells treated with CT- 1 showed better organized sacromeric structure and more mitochondria, which are morphological characteristic of matured cardiomyocytes. Western blot demonstrated that CT-1 promotes cardiomyocyte differentiation from miPSCs partly via JAK2/STAT3/Pim-1 pathway as compared with control group. Conclusions These findings suggested that CT-1 could enhance the cardiomyocyte differentiation as well as the maturation of mouse induced pluripotent stem cell derived cardiomyocytes by regulating JAK2/STAT3/Pim-1 signaling pathway.展开更多
The purpose of the present study was to compare the effects of different doses of ultraviolet radiation A1 （UVA1） on human fibroblast proliferation and collagen level in a mouse model of scleroderma, so as to identi...The purpose of the present study was to compare the effects of different doses of ultraviolet radiation A1 （UVA1） on human fibroblast proliferation and collagen level in a mouse model of scleroderma, so as to identify appropriate irradiation doses for clinical treatment of scleroderma. Monolayer from human fibroblasts was cultured in vitro, and a mouse model of scleroderma was established by subcutaneous injection of 100 μL of 400 μg/mL bleomycin into the back of BALB/c mice for 4 weeks. The mouse models and human fibroblasts were divided into UVA1- exposed （100, 60 and 20 J/cm2） and UVA-unexposed groups. At 0, 24 and 48 h after exposure, cell proliferation and levels of hydroxyproline and collagen were detected. UVA1 irradiation was performed 3 times weekly for 10 weeks, and the pathological changes of skin tissues, skin thickness and collagen level were observed after phototherapy. Cell proliferation and the levels of hydroxyproline and collagen were inhibited after phototherapy, and there was a significant difference between the UVAl-exposed cells and UVAl-unexposed cells （P 〈 0.001）. In addition, UVA1 phototherapy improved dermal sclerosis and softened the skin, and there were significant differences between the high-dose UVA1 group and the model group, and the negative group （P 〈 0.05）. It is concluded that UVA1 radiation can reduce cell proliferation, and decrease hydroxyproline and collagen levels in a dose-dependent manner in vitro. High-dose UVA1 phototherapy has marked therapeutic effect on scleroderma in the mouse model. Decreased collagen level may be related to the reduced number and activity of cells, as well as inhibition of collagen synthesis.展开更多
Coxsackievirus B1(CVB1) is a leading causative agent of severe infectious diseases in humans and has been reported to be associated with outbreaks of aseptic meningitis, myocarditis, and the development of chronic dis...Coxsackievirus B1(CVB1) is a leading causative agent of severe infectious diseases in humans and has been reported to be associated with outbreaks of aseptic meningitis, myocarditis, and the development of chronic diseases such as type 1 diabetes mellitus(T1DM). There is no approved vaccine or effective antiviral therapy to treat CBV1 infection. And animal models to assess the effects of antiviral agents and vaccine remain limited. In this study, we established a neonatal mouse model of CVB1 using a clinically isolated strain to characterize the pathological manifestations of virus infection and to promote the development of vaccines and antiviral drugs against CVB1. One-day-old BALB/c mice were susceptible to CVB1 infection by intraperitoneal injection. Mice challenged with CVB1 at a low dose [10 median tissue culture infective dose(TCID_(50))] exhibited a series of clinical symptoms, such as inactivity, emaciation, limb weakness, hair thinning,hunching and even death. Pathological examination and tissue viral load analysis showed that positive signals of CVB1 were detected in the heart, spinal cord, limb muscle and kidney without pathological damage. Particularly, CVB1 had a strong tropism towards the pancreas, causing severe cellular necrosis with inflammatory infiltration, and was spread by viraemia. Notably, the monoclonal antibody(mAb) 6H5 and antisera elicited from CVB1-vaccinated mice effectively protected the mice from CVB1 infection in the mouse model. In summary, the established neonatal mouse model is an effective tool for evaluating the efficacy of CVB1 antiviral reagents and vaccines.展开更多
To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and e...To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and expressions of MMP-2 and MMP-9 were analyzed with zymography in a TGF-β1 transgenic mouse model after thoracic irradiation with 12 Gy. We found expressions of TGF-β1 in the lung from the transgenic mice were three folds as compared to those from control mice. With densitometrical analysis, we found a significant decrease in MMP-9 activity in lung homogenates from the transgenic mice as compared with those from non-transgenic control mice 8 weeks after sham-irradiation （relative MMP-9 activity： C： 1. 000±0. 1091; TG： 0. 4772±0. 470 （n=8, P〈0.05）. But MMP-2 was constitutively expressed in the lung homogenates from the transgenic mice as compared to those from control mice 8 weeks after sham-irradiation （relative MMP-2 activity 8 weeks after sham-irradiation： C： 1. 000±0. 1556, TG： 1. 0075±0. 1472）. Eight weeks after thoracic irradiation with 12 Gy, we observed a significant increase of MMP-2 and MMP-9 activity in lung homogenates from both transgenic and normal mice. In TGF-β1 transgenic mice relative MMP-9 activity was increased to 1. 5321±0. 2217 folds 8 weeks after thoracic irradiation with 12 Gy as compared to those after sham-irradiation （1. 000±0. 2153）, and relative MMP-2 activity was increased to 1. 7142 ± 0. 4231 folds. Our results show that TGF-β1 itself down-regulates activity of MMP-9, thereby decreases ECM degradation in lungs of TGF-β1 transgenic mice. Also we find that ionizing irradiation upregulates both MMP-2 and MMP-9 activity. Over-expressions of MMP-9 and MMP-2 after lung irradiation are involved in the inflammatory response associated with radiation-induced lung injury, and maybe further in radiation-induced lung fibrosis.展开更多
Aim： To investigate the expression of Spindlin 1 （Spin First, reverse-transcription polymerase chain reaction present in mouse testis. Then the expression patterns 1） isoform2 and assess its function in mouse testi...Aim： To investigate the expression of Spindlin 1 （Spin First, reverse-transcription polymerase chain reaction present in mouse testis. Then the expression patterns 1） isoform2 and assess its function in mouse testis. Methods： （RT-PCR） was used to determine whether Spinl isoform2 is of the isoform between newborn and adult mice testes were compared by immunoblot analysis. Finally, the diversity of its localization in mice testes at different ages （days 0, 7, 14, 21, 28 and 60） was observed by immunohistochemistry. The localization of the protein in mouse sperm was also investigated by immunofluorescence. Results： The RT-PCR results show that Spinl isoform2 is present in mouse testis. As shown by immunoblot analysis, the isoform was more highly expressed in adult testes compared with newborn testes. Interestingly, Spinl isoform2 did not show up in the cytoplasm of primary spermatocytes until day 14. Also, the protein exists at the tail of the mouse sperm. Conclusion： Spinl isoform2 is a protein expressed highly in adult testis, which might be involved in spermatogenesis and could be necessary for normal sperm motility.展开更多
基金supported by the National Natural Science Foundation of China (32471049,32170984,32471188,32200802)Natural Science Foundation of Shandong Province (ZR2023QH110)。
文摘Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.
文摘目的:探讨同型半胱氨酸(Hcy)促进单核细胞趋化蛋白-1(MCP-1)表达诱导巨噬细胞浸润加重急性缺血性卒中(AIS)的机制。方法:采用线栓法制备局灶性脑缺血再灌注小鼠模型。采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑缺血严重程度,HE染色法检测缺血脑组织神经细胞损伤状况和炎症细胞浸润程度;高效液相色谱法检测急性期小鼠脑组织Hcy表达;免疫组织化学染色、Western blot、real time RT-PCR法检测小鼠脑组织MCP-1表达。Transwell小室检测巨噬细胞迁移程度,TUNEL法检测Hcy对内皮细胞影响;利用MCP-1中和抗体观察对巨噬细胞迁移的影响。结果:与假手术组比较,模型组小鼠右侧大脑半球梗死明显,缺血脑组织损伤严重,神经细胞死亡并伴有大量炎症细胞浸润;Hcy水平显著升高,MCP-1蛋白和mRNA表达均明显增加。Hcy刺激小鼠脑血管内皮细胞后可诱导NF-κB(p65)入核,促进巨噬细胞迁移,促进内皮细胞凋亡,给予NF-κB(p65)抑制剂后MCP-1生成减少;MCP-1中和抗体明显抑制巨噬细胞迁移。结论:Hcy、MCP-1与脑缺血再灌注炎性反应相关,且Hcy可能通过NF-κB通路促进MCP-1表达,发挥诱导巨噬细胞浸润作用,加重脑缺血损伤。
文摘目的探究人乳腺癌裸鼠移植动物模型体内孕激素受体膜组分1(PGRMC1)在黄体酮与炔诺酮(NET)刺激下对乳腺癌异种移植瘤增殖影响的差异性。方法构建PGRMC1稳定表达的人乳腺癌细胞系(MCF7),36只雌性BALB/c-nu裸鼠按随机数字表法分为MCF7/HA-PGRMC1组(n=18)和MCF7/HA-Vector组(n=18)。各鼠于颈部皮下包埋雌二醇(E_(2))缓释片。24 h后两组裸鼠左、右侧面皮下分别注射MCF7/HA-PGRMC1和MCF7/HA-Vector细胞悬液150μL,注射细胞数为1×107个。肿瘤生长至100 mm 3左右,将两组裸鼠进行组内分组,分别皮下联合包埋安慰剂、黄体酮缓释片、NET缓释片,即E_(2)+安慰剂亚组(n=6)、E_(2)+黄体酮亚组(n=6)、E_(2)+NET亚组(n=6)。记录肿瘤体积变化,绘制肿瘤生长曲线。45 d后完整摘除肿瘤组织免疫组化染色检测PGRMC1和Ki67表达情况。结果MCF7/HA-PGRMC1株系构建成功,在MCF7/HA-PGRMC1组中,与E_(2)+安慰剂亚组相比,E_(2)+黄体酮亚组肿瘤增长差异无统计学意义(P>0.05),而E_(2)+NET亚组肿瘤明显生长(P<0.05)。在MCF7/HA-Vector组中,与E_(2)+安慰剂亚组相比,E_(2)+NET亚组、E_(2)+黄体酮亚组的肿瘤增长差异均无统计学意义(P>0.05)。在MCF7/HA-PGRMC1组中,E_(2)+NET亚组Ki67呈强阳性表达,PGRMC1与Ki67表达呈正相关(r=0.892,P=0.014)。结论该实验在裸鼠体内证实PGRMC1过表达可特异性介导NET联合E_(2)的促乳腺癌增殖效应,而联合黄体酮并没有促进额外增殖。这一发现为临床激素治疗中孕激素类型的选择提供了关键的动物实验依据。
基金Supported by National Natural Science Foundation of China,No.30940069the Natural Sciences Foundation of Beijing,No.7102127
文摘AIM: To characterize high-mobility group protein 1-toll-like receptor 4(HMGB1-TLR4) and downstream signaling pathways in intestinal ischemia/reperfusion(I/R) injury.METHODS: Forty specific-pathogen-free male C57BL/6 mice were randomly divided into five groups(n = 8 per group): sham, control, anti-HMGB1, anti-myeloid differentiation gene 88(My D88), and anti-translocatingchain-associating membrane protein(TRIF) antibody groups. Vehicle with the control Ig G antibody, antiHMGB1, anti-My D88, or anti-TRIF antibodies(all 1 mg/kg, 0.025%) were injected via the caudal vein 30 min prior to ischemia. After anesthetization, the abdominal wall was opened and the superior mesenteric artery was exposed, followed by 60 min mesenteric ischemia and then 60 min reperfusion. For the sham group, the abdominal wall was opened for 120 min without I/R. Levels of serum nuclear factor(NF)-κB p65, interleukin(IL)-6, and tumor necrosis factor(TNF)-α were measured, along with myeloperoxidase activity in the lung and liver. Inaddition,morphologic changes that occurred in the lung and intestinal tissues were evaluated. Levels of m RNA transcripts encoding HMGB1 and NF-κB were measured by real-time quantitative PCR, and levels of HMGB1 and NF-κB protein were measured by Western blot. Results were analyzed using one-way analysis of variance.RESULTS: Blocking HMGB 1, MyD 8 8, and TRIF expression by injecting anti-HMGB1, anti-My D88, or anti-TRIF antibodies prior to ischemia reduced the levels of inflammatory cytokines in serum; NF-κB p65: 104.64 ± 11.89, 228.53 ± 24.85, 145.00 ± 33.63, 191.12 ± 13.22, and 183.73 ± 10.81(P < 0.05); IL-6: 50.02 ± 6.33, 104.91 ± 31.18, 62.28 ± 6.73, 85.90 ± 17.37, and 78.14 ± 7.38(P < 0.05); TNF-α, 43.79 ± 4.18, 70.81 ± 6.97, 52.76 ± 5.71, 63.19 ± 5.47, and 59.70 ± 4.63(P < 0.05) for the sham, control, anti-HMGB1, anti-My D88, and anti-TRIF groups, respectively(all in pg/m L).Antibodies also alleviated tissue injury in the lung and small intestine compared with the control group in the mouse intestinal I/R model. The administration of antiHMGB1, anti-My D88, and anti-TRIF antibodies markedly reduced damage caused by I/R, for which anti-HMGB1 antibody had the most obvious effect.CONCLUSION: HMGB1 and its downstream signaling pathway play important roles in the mouse intestinal I/R injury, and the effect of the TRIF-dependent pathway is slightly greater.
基金supported by the National Nature Science Foundation of China(No.81302772)Jiangsu Provincial Medical Talent Project(No.QNRC2016716)
文摘The present study was designed to elucidate whether the mechanism by which osthole decreases collagenⅠ/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts(CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pc DNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·m L^(-1) of osthole for 24 h, the m RNA and protein expression levels of collagensⅠand III were reduced. The collagen Ⅰ/III ratio was also reduced. The m RNA and protein expression levels of TGF-β1, TβRⅠ, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen Ⅰ and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.
基金supported by funding from Innovative Research Team in Nanjing Medical University(IRT0631)the collaborating Grants(30611120524)
文摘Objective： Recent studies have shown that the local expression of soluble interleukin （IL） -1 receptor type Ⅱ （slL-1 R Ⅱ ） in endometrial tissue of women with endometriosis is decreased, and the depression of IL-1 R Ⅱ was more significant in infertile women than that in fertile women with endometriosis. In this research, we investigated the remedial effect of slL-1-R Ⅱ administration on endometriosis in the nude mouse model. Methods： Nineteen nude model mice with endometriosis were randomly divided into three groups： group A was treated by intraperitoneal administration with only slL-1 R Ⅱ for two weeks, group B was similarly treated with only IL- 1, and group C （control） was administered saline. After 2 weeks, the size of the ectopic endometrial lesions was calculated, and the expression of vascular endothelial growth factor （VEGF） and B-cell lymphoma leukemia-2 （Bcl- 2） were detected by immunohistochemistry. The IL-8 and VEGF levels in the peritoneal fluid （PF） and serum were also measured by enzyme-linked immunosorbent assay （ELISA）. Results： The mean size of ectopic endometrial lesion did not differ between the three groups （P 〉 0.05）. Compared with the control, the expression of VEGF and Bcl-2 was significantly lower in group A, and higher in group B. In the three groups, the levels of IL-8 in the PF and serum were highest in group A, and lowest in group B. Conclusion： slL-1 R Ⅱ may suppresse hyperplasia of ectopic endometriosis, perhaps by reducing the expression of certain cytokines, such as VEGF, IL-8, and Bcl-2, which could provide a new clinical strategy for the treatment of endometriosis.
文摘This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups： control group, D2 group （2 days after pregnancy）, D4 group （4 days after pregnancy）, D6 group （6 days after pregnancy） and D8 group （8 days after pregnancy）, each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups （P〈0.01）. Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy （day 2）, and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice （P〈0.01）. In situ hybridization showed Ang-I mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 （P〈0.01）. It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.
基金This work was supported by the National Funds for Distinguished Young Scientists of China （No. 81325009） and National Nature Science Foundation of China （No. 81270168, No. 81227901）, （Feng Cao BWS12J037）, Innovation Team granted by Ministry of Education PRC （IRT1053）, National Basic Research Program of China （2012CB518101）. Shaanxi Province Program （2013K12-02-03, 2014KCT-20）. The authors declare no conflict of interest.
文摘Background The induced pluripotent stem cell （iPSC） has shown great potential in cellular therapy of myocardial infarction （MI）, while its application is hampered by the low efficiency of cardiomyocyte differentiation. The present study was designed to investigate the effects of cardiotrophin-1 （CT-1） on cardiomyocyte differentiation from mouse induced pluripotent stem cells （miPSCs） and the underlying mechanisms involved. Methods The optimal treatment condition for cardiomyocyte differentiation from miPSCs was established with ideal concentration （10 ng/mL） and duration （from day 3 to day 14） of CT-1 administration. Up-regulated expression of cardiac specific genes that accounted for embryonic cardiogenesis was observed by quantitative RT-PCR. Elevated amount of a-myosin heavy chain （ct-MHC） and cardiac troponin I （cTn I） positive cells were detected by immunofluorescence staining and flow cytometry analysis in CT- 1 group. Results Transmission electron microscopic analysis revealed that cells treated with CT- 1 showed better organized sacromeric structure and more mitochondria, which are morphological characteristic of matured cardiomyocytes. Western blot demonstrated that CT-1 promotes cardiomyocyte differentiation from miPSCs partly via JAK2/STAT3/Pim-1 pathway as compared with control group. Conclusions These findings suggested that CT-1 could enhance the cardiomyocyte differentiation as well as the maturation of mouse induced pluripotent stem cell derived cardiomyocytes by regulating JAK2/STAT3/Pim-1 signaling pathway.
文摘The purpose of the present study was to compare the effects of different doses of ultraviolet radiation A1 （UVA1） on human fibroblast proliferation and collagen level in a mouse model of scleroderma, so as to identify appropriate irradiation doses for clinical treatment of scleroderma. Monolayer from human fibroblasts was cultured in vitro, and a mouse model of scleroderma was established by subcutaneous injection of 100 μL of 400 μg/mL bleomycin into the back of BALB/c mice for 4 weeks. The mouse models and human fibroblasts were divided into UVA1- exposed （100, 60 and 20 J/cm2） and UVA-unexposed groups. At 0, 24 and 48 h after exposure, cell proliferation and levels of hydroxyproline and collagen were detected. UVA1 irradiation was performed 3 times weekly for 10 weeks, and the pathological changes of skin tissues, skin thickness and collagen level were observed after phototherapy. Cell proliferation and the levels of hydroxyproline and collagen were inhibited after phototherapy, and there was a significant difference between the UVAl-exposed cells and UVAl-unexposed cells （P 〈 0.001）. In addition, UVA1 phototherapy improved dermal sclerosis and softened the skin, and there were significant differences between the high-dose UVA1 group and the model group, and the negative group （P 〈 0.05）. It is concluded that UVA1 radiation can reduce cell proliferation, and decrease hydroxyproline and collagen levels in a dose-dependent manner in vitro. High-dose UVA1 phototherapy has marked therapeutic effect on scleroderma in the mouse model. Decreased collagen level may be related to the reduced number and activity of cells, as well as inhibition of collagen synthesis.
基金This research was supported by grants from the National Natural Science Foundation of China(No.82072282 and 81801646)the National Science and Technology Major Project of Infectious Diseases(No.2017ZX10304402-002-003)the National Science and Technology Major Projects for Major New Drugs Innovation and Development(No.2018ZX09711003-005003)。
文摘Coxsackievirus B1(CVB1) is a leading causative agent of severe infectious diseases in humans and has been reported to be associated with outbreaks of aseptic meningitis, myocarditis, and the development of chronic diseases such as type 1 diabetes mellitus(T1DM). There is no approved vaccine or effective antiviral therapy to treat CBV1 infection. And animal models to assess the effects of antiviral agents and vaccine remain limited. In this study, we established a neonatal mouse model of CVB1 using a clinically isolated strain to characterize the pathological manifestations of virus infection and to promote the development of vaccines and antiviral drugs against CVB1. One-day-old BALB/c mice were susceptible to CVB1 infection by intraperitoneal injection. Mice challenged with CVB1 at a low dose [10 median tissue culture infective dose(TCID_(50))] exhibited a series of clinical symptoms, such as inactivity, emaciation, limb weakness, hair thinning,hunching and even death. Pathological examination and tissue viral load analysis showed that positive signals of CVB1 were detected in the heart, spinal cord, limb muscle and kidney without pathological damage. Particularly, CVB1 had a strong tropism towards the pancreas, causing severe cellular necrosis with inflammatory infiltration, and was spread by viraemia. Notably, the monoclonal antibody(mAb) 6H5 and antisera elicited from CVB1-vaccinated mice effectively protected the mice from CVB1 infection in the mouse model. In summary, the established neonatal mouse model is an effective tool for evaluating the efficacy of CVB1 antiviral reagents and vaccines.
文摘To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and expressions of MMP-2 and MMP-9 were analyzed with zymography in a TGF-β1 transgenic mouse model after thoracic irradiation with 12 Gy. We found expressions of TGF-β1 in the lung from the transgenic mice were three folds as compared to those from control mice. With densitometrical analysis, we found a significant decrease in MMP-9 activity in lung homogenates from the transgenic mice as compared with those from non-transgenic control mice 8 weeks after sham-irradiation （relative MMP-9 activity： C： 1. 000±0. 1091; TG： 0. 4772±0. 470 （n=8, P〈0.05）. But MMP-2 was constitutively expressed in the lung homogenates from the transgenic mice as compared to those from control mice 8 weeks after sham-irradiation （relative MMP-2 activity 8 weeks after sham-irradiation： C： 1. 000±0. 1556, TG： 1. 0075±0. 1472）. Eight weeks after thoracic irradiation with 12 Gy, we observed a significant increase of MMP-2 and MMP-9 activity in lung homogenates from both transgenic and normal mice. In TGF-β1 transgenic mice relative MMP-9 activity was increased to 1. 5321±0. 2217 folds 8 weeks after thoracic irradiation with 12 Gy as compared to those after sham-irradiation （1. 000±0. 2153）, and relative MMP-2 activity was increased to 1. 7142 ± 0. 4231 folds. Our results show that TGF-β1 itself down-regulates activity of MMP-9, thereby decreases ECM degradation in lungs of TGF-β1 transgenic mice. Also we find that ionizing irradiation upregulates both MMP-2 and MMP-9 activity. Over-expressions of MMP-9 and MMP-2 after lung irradiation are involved in the inflammatory response associated with radiation-induced lung injury, and maybe further in radiation-induced lung fibrosis.
基金This research was supported by grants from the 973 Program （2006CB504002, 2006CB944002） and the National Natural Science Foundation of China （30425006）.
文摘Aim： To investigate the expression of Spindlin 1 （Spin First, reverse-transcription polymerase chain reaction present in mouse testis. Then the expression patterns 1） isoform2 and assess its function in mouse testis. Methods： （RT-PCR） was used to determine whether Spinl isoform2 is of the isoform between newborn and adult mice testes were compared by immunoblot analysis. Finally, the diversity of its localization in mice testes at different ages （days 0, 7, 14, 21, 28 and 60） was observed by immunohistochemistry. The localization of the protein in mouse sperm was also investigated by immunofluorescence. Results： The RT-PCR results show that Spinl isoform2 is present in mouse testis. As shown by immunoblot analysis, the isoform was more highly expressed in adult testes compared with newborn testes. Interestingly, Spinl isoform2 did not show up in the cytoplasm of primary spermatocytes until day 14. Also, the protein exists at the tail of the mouse sperm. Conclusion： Spinl isoform2 is a protein expressed highly in adult testis, which might be involved in spermatogenesis and could be necessary for normal sperm motility.
