High-fat diet (HFD), characterized by an increased proportion of palmitic acid (PA), can induce inflammatory responses in Sertoli cells, trigger apoptosis, and cause spermatogenic dysfunction. Endoplasmic reticulum stress (ERS) is intimately linked to inflammation, but whether ERS contributes to HFD-induced Sertoli cell inflammatory injury remains unclear. This study investigated how HFD and PA mediated Sertoli cell injury by upregulating inflammatory response at the in vivo and cellular levels. Obese mice and TM4 cell models were established using HFD and PA, respectively. The sperm quality of mice was systematically assessed. Key regulatory pathways were identified via transcriptome sequencing, and the mechanism underlying HFD-induced Sertoli cell injury was validated using Western blot and flow cytometry. The results showed that HFD could upregulate the expression of inflammatory cytokines in Sertoli cells by activating the NF-κB signaling pathway, ultimately leading to a reduction in Sertoli cell numbers. Through transcriptome sequencing, we found that PA could activate ERS in TM4 cells. After inhibiting the activity of ERS transmembrane protein inositol-requiring 1α (IRE1α), the apoptosis rate, inflammatory cytokine production, and the expression levels of NF-κB signaling pathway proteins in PA-stimulated TM4 cells were significantly decreased. Notably, inhibition of IRE1α protein activity significantly downregulated the expression of adaptor protein tumor necrosis factor 2 (TRAF2). Knocking down TRAF2 reduced the expression of both NF-κB signaling pathway proteins and inflammatory cytokines. Overall, this study provides a theoretical basis for preventing and treating HFD-induced male reproductive dysfunction by targeting the IRE1α/ TRAF2/NF-κB axis.

Di-(2-ethylhexyl) phthalate (DEHP), a widely used plasticizer, could cause male reproductive toxicity by disrupting spermatogenesis. Piwi-interacting RNAs (piRNAs) are a small non-coding RNAs specifically highly expressed in the germline and interact with PIWI proteins to regulate spermatogenesis. Accumulating studies have confirmed that environmental poisons could induce male reproductive injury via altering piRNA expression. However, it remains unclear whether DEHP causes male reproductive dysfunction by perturbing piRNA expression levels. In this study, we conducted piRNA microarray expression analyses on testes of DEHP-exposed and control male rats and performed some in vitro and in vivo studies to explore the role of piRNA on DEHP-induced male reproductive toxicity. Our results showed that DEHP exposure leaded to changed expression profiles of piRNAs in pubertal male rat testes. And bioinformatics analyses revealed that down-regulated piR-rno-26751 probably targeted Insr mRNA expression regulation. Results from gene and protein expression tests demonstrated that DEHP caused decreased expression level of INSR mainly in spermatogonia. Moreover, MEHP, the main metabolite of DEHP resulted in cell apoptosis and down-regulation of INSR and its downstream p-IRS1, p-PI3K, p-AKT and p-FOXO1 in GC-1spg cells. Conversely, overexpression of INSR restored cell apoptosis and the down-regulation of the above proteins in GC-1spg cells. In conclusion, these findings suggest that DEHP-induced down-regulation of piR-rno-26751 targets the suppression of INSR, leading to apoptosis of spermatogonia in pubertal male rats.

Cadmium (Cd) is a harmful environmental pollutant that disrupts public health, including respiratory, digestive, and reproductive systems. In this study, male rats were exposed to CdCl2 at a dose of 3 mg/kg by oral for 28 days to investigate the impact on spermatogenesis. Testis tissue samples were collected after sacrifice, and piRNA expression levels were measured using piRNA microarray and qPCR. PiRNAs, specialized molecules involved in spermatogenesis, were examined. CdCl2 exposure led to disrupted piRNA expression, particularly in piRNADQ759395 in rats. This piRNA was found to have a binding site with p53, and a similar piRNA-DQ717867 was discovered in mice. In GC-2spd cells, CdCl2 exposure increased piRNA-DQ717867 expression, which resulted in cell cycle arrest and abnormal expression of cell cycle-related proteins. The activation of p53-related pathways and disruptions in cell cycle regulation were also observed. Antagomir-717867 transfections and PFT-a pretreatment in GC-2spd cells supported the involvement of piRNA-DQ717867 in regulating cell cycle-related proteins. This study suggests that Cd exposure induces abnormal expression of piRNA-DQ759395 in rat testis and that piRNA-DQ717867 may regulate p53, causing cell cycle abnormalities in GC-2spd cells. These findings help understand the mechanisms of male reproductive toxicity caused by Cd exposure and emphasize the role of piRNAs in cell cycle regulation and male reproductive health.

目的 从内质网应激(endoplasmic reticulum stress,ERS)角度探讨棕榈酸(palmitic acid,PA)诱导小鼠精原细胞株GC-1 细胞凋亡的机制.方法 将GC-1 细胞分为正常组(Control)、溶剂对照组(Vehicle)和不同浓度 PA 处理组(100、150、200、250 μmol·L-1).MTT法检测细胞增殖活力,流式细胞术Annexin V-FITC/PI双染法检测细胞凋亡情况;Caspaes-3 活性检测试剂盒检测凋亡蛋白caspase-3 的活性;Western blot检测凋亡相关蛋白和ERS 相关蛋白表达水平.结果 PA处理GC-1 细胞 48h后,细胞增殖活力显著下降,细胞凋亡率上升,凋亡蛋白caspase-3 活性增高,结果均具有浓度依赖性;PA可明显增加 GC-1 细胞促凋亡蛋白cleaved-caspase-3 的表达水平,明显降低抑凋亡蛋白BCL2 的表达水平,同时上调 ERS 相关蛋白 GRP78、CHOP、p-IRE1、XBP1 的表达水平,但对 p-PERK、PERK、p-eIF2α、eIF2α、ATF4 以及ATF6 的表达无显著影响.结论 PA 诱导小鼠GC-1 细胞凋亡可能与激活IRE1 通路有关.

Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplas-mic reticulum stress.Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly,with six mice in each group.The mice in normal group or high-fat diet group were fed with regu-lar or high-fat diet continuously for five months.The mice were weighed,anesthetized,and euthanized to col-lect testicular and epididymal tissue for analysis.The testicular tissue was weighed and their indices were calculated.Epididymal tissue was collected for semen analysis.The morphological alterations of testicular tis-sue were observed using hematoxylin-eosin(HE)stai-ning.The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calcu-lated.The expression levels of apoptosis and endoplas-mic reticulum stress-related proteins in testicular tissue were detected by Western blot.The protein expression and localization of GRP78 in testicular tissue were fur-ther detected by immunofluorescence.Results The results showed that compared to the normal group,the high-fat diet group had a significant increase in body weight,a significant decrease in testicular index,sperm concentration,and sperm vability,loose arrangement of germ cells,significant thinning of the seminiferous epi-thelium,no significant change in the diameter of semi-niferous tubules,a significant increase in germ cell ap-optosis,with an increased apoptosis index,and signifi-cant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expres-sion.The expression levels of GRP78,p-IRE1,XBP1,and ATF6α proteins were significantly up-regulated,while p-PERK,p-eIF2α,ATF4 protein expression showed no significant changes.Immunofluorescence re-sults further showed a significant increase in the ex-pression of GRP78 protein in the testicular tissue,with no significant changes in the expression location.Con-clusions High-fat diet can induce the apoptosis of mouse testicular germ cells,and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.

Perfluorooctanoic acid (PFOA), a man-made chemical widely used in consumers, could cause male reproductive toxicity by disrupting blood-testis barrier (BTB) integrity. Autophagy in Sertoli cells is essential for regulation of spermatogenesis and BTB. However, it remains a mystery that whether PFOA-induced BTB injury is associated with autophagy in Sertoli cells. In this study, we found that PFOA dose-dependently disrupted tight junction (TJ) function in Sertoli cells in vivo and in vitro. Furthermore, the results from transmission electron microscopy, Western blot and immunofluorescence analysis revealed that PFOA induced the accumulation of autophagosome in testicular Sertoli cells as well as TM4 cells. Further study confirmed that autophagosome accumulation resulted from the blockage of autophagic degradation because of disruption of autophagosome and lysosome fusion via downregulation of the expression of α-SNAP. In parallel, the overexpressed MMP9 was also observed in vivo and in vitro. Conversely, overexpression of α-SNAP inhibited the expression of MMP9 in TM4 cells. In conclusion, PFOA blocks autophagic flux through downregulating the expression levels of α-SNAP in Sertoli cells, and then induces the accumulation of MMP9 leading to disruption of TJ function. This finding will provide clues for effective prevention and treatment of PFOA-induced male reproductive toxicity.

OBJECTIVE:To study the impact of cadmium (Cd) on the expressions of PIWI-interacting RNAs (piRNA) in the rat testis and its possible action mechanism.METHODS:Twelve 6-week-old SD rats were randomly divided into a Cd-exposure and a control group, the former gavaged with CdCl2 at 3 mg/kg/d and the latter with normal saline, all for 28 successive days. Then the testicular tissues were collected from the rats, sperm concentration and motility were obtained by computer-assisted sperm analysis (CASA), and piRNA sequencing was performed using the gene chip, followed by bioinformatics analysis of differentially expressed piRNAs.RESULTS:Compared with the controls, the rats in the Cd-exposure group showed significantly decreased sperm concentration and motility (P < 0.05). The expressions of 272 piRNAs were up-regulated and 402 down-regulated after 28 days of Cd exposure, and 4 of the up-regulated piRNAs were consistent with the results of gene chip verification. Bioinformatics analysis showed that the 4 up-regulated piRNA target genes were involved in 50 biological processes, such as negative regulation of apoptosis, positive regulation of gene expression and positive regulation of GTPase activity, and mainly concentrated in 13 signaling pathways including transcription dysregulation, calcium and mitogen-activated protein kinase signaling pathways in cancer. Among them, PIRNA-DQ765261 had a binding site with Bcl-2.CONCLUSION:Cadmium can induce changes in the expressions piRNAs in the rat testicular tissue, and some piRNAs may be involved in the autophagy and apoptosis of sperm. Bcl-2 may be the target of PIRNA-DQ765261.

目的 研究氧化应激在邻苯二甲酸单(2-乙基己基)酯(mono-2-ethylhexyl-phthalate,MEHP)诱导小鼠初级精母细胞(GC-2spd)毒性中的作用及机制.方法 将培养的GC-2spd细胞分为对照组(含0.1％二甲基亚砜的无血清培养基)、MEHP染毒组(1、10、100 μmol/L MEHP 3个不同剂量组)、N-乙酰半胱氨酸(N-acetylcysteine,NAC)预处理组(100 μmol/L NAC+100 μmol/L MEHP),MEHP 染毒24h,NAC预处理2h,NAC的预处理浓度(100 μmol/L)通过CCK-8实验确定.采用DCFH-DA探针法检测细胞内活性氧(reactive oxide species,ROS)水平,JC-1法检测线粒体膜电位,Western blot法检测细胞内线粒体凋亡通路相关蛋白和p-STAT3Tyr705、P53蛋白表达水平.结果 随着MEHP染毒剂量升高,细胞内代表活性氧水平的绿色荧光信号增多;不同剂量MEHP染毒组的线粒体膜电位均下降,表现为红绿荧光细胞比例比值减小,10和100μmol/L MEHP染毒组的红绿荧光细胞比例比值分别为5.15±1.68和4.09±1.72,明显低于对照组(7.91±1.24)(P＜0.05);促凋亡蛋白Bax与抗凋亡蛋白Bcl-2表达水平比值(Bax/Bcl-2)在1、10、100 μmol/L MEHP染毒组分别为 1.23±0.17、2.64±0.43和4.75±0.73,与对照组(0.52±0.11)相比明显增加(P＜0.05);cytochrome C 蛋白表达水平在 100 μmol/L MEHP 染毒组升高为0.83±0.09,cleaved caspase-9蛋白表达水平在10和100 μmol/L MEHP染毒组升高为0.41±0.03和0.52±0.09,cleaved caspase-3蛋白表达水平在 10和 100 μmol/L MEHP 染毒组升高为0.60±0.12和0.84±0.17,与对照组相比差异均具有统计学意义(P＜0.05);STAT3/p53通路中,p-STAT3Tyr705蛋白表达水平在10和100 μmol/L MEHP染毒组分别下降为0.70±0.14和0.41±0.04,P53蛋白表达水平在10和100 μmol/L MEHP染毒组分别升高为1.32±0.05和1.66±0.22,与对照组相比差异均具有统计学意义(P＜0.05);NAC预处理后,与100 μmol/L MEHP组相比,NAC预处理组ROS生成减少,线粒体膜电位升高为5.92±1.64,Bax/Bcl-2下降为 1.92±0.06,cytochrome C、cleaved caspase-9和 cleaved caspase-3蛋白表达水平分别下降为0.57±0.07、0.35±0.04和0.53±0.06,p-STAT3Tyr705蛋白表达水平升高为0.86±0.07,P53蛋白表达水平下降为1.01±0.06,差异均有统计学意义(P＜0.05).结论 MEHP诱导GC-2spd细胞凋亡可能与ROS介导的STAT3/p53通路调控线粒体凋亡通路有关.

Lung cancer is the leading cause of malignancy-related mortality and lung adenocarcinoma accounts for about 40% of lung malignancies. The aim of this study was to investigate the associations of intraflagellar transport protein 20 (IFT20) and Golgi matrix protein 130 (GM130) expression with clinicopathological features and survival in patients with lung adenocarcinoma. The expressions of IFT20 and GM130 protein in cancerous and matched adjacent lung tissues of 235 patients with lung adenocarcinoma were assessed by tissue microarray and immunohistochemistry, which were indicated by the mean optical density (IOD/area), the rate of positive staining cells and staining intensity score. The correlation between IFT20 and GM130 protein was assessed by Spearman’s rank correlation. Associations of IFT20 and GM130 protein expression with clinicopathological features of patients were analyzed by multivariate logistic regression models. The survival analysis of patients was performed by Cox proportional hazard regression models. With adjustment for multiple potential confounders, each one-point increase in IFT20 protein staining intensity score was significantly associated with 32% and 29% reduced risk for TNM stage in II ~ IV and lymphatic metastasis of patients, respectively (P < 0.05). And each one-point increase in GM130 protein staining intensity score was associated with a significant reduction in the risk of poor differentiation and tumors size > 7 cm by 29% and 38% for lung adenocarcinoma patients, respectively (P < 0.05). In stratified Cox model analysis, enhanced IFT20 staining intensity score was significantly decreased the risk of death by 16% for patients without distant metastasis. And elevated the IOD/area of GM130 expression significantly decreased the death risk of lung adenocarcinoma patients with tumor size > 7 cm or distant metastasis by 54% and 65%, respectively (P < 0.05). IFT20 and GM130 protein expressions were negatively associated with tumor differentiated types, size, TNM stage and lymphatic metastasis of lung adenocarcinoma. Both IFT20 and GM130 proteins have some protective effects on the survival of lung adenocarcinoma patients with specific clinicopathological features.

Cadmium (Cd) is a heavy metal that is widely present in modern industrial production. It is a known, highly toxic environmental endocrine disruptor. Long-term exposure to Cd can cause varying degrees of damage to the liver, kidney, and reproductive system of organisms, especially the male reproductive system. This study aimed to explore the mechanism of Cd toxicity in the male reproductive system during puberty. Eighteen healthy 6-week-old male Sprague-Dawley rats were randomly divided into three groups (control group, low-dose group, and high-dose group) according to their body weight, with six in each group. Cd (0, 1, and 3 mg/kg/day) was given by gavage for 28 consecutive days. The results showed that Cd exposure to each dose group caused a decrease in the testicular organ coefficient and sperm count, compared with the control group. Cd exposure resulted in significant changes in testicular morphology in the 3 mg/kg/day Cd group. In the 1 and 3 mg/kg/day Cd groups, serum testosterone decreased and apoptosis of testicular cells increased significantly (p < 0.05). In addition, compared with the control group, the activity of glutathione peroxidase and superoxide dismutase in each Cd exposure dose group decreased, but the content of malondialdehyde in the high-dose, 3 mg/kg/day Cd treatment group significantly increased (p < 0.05). Although Cd exposure caused an increase in the messenger RNA (mRNA) levels of Bcl-2, Caspase-3 and Caspase-9 in the testicular tissues (p < 0.05), Bcl-2 expression was unchanged (p > 0.05). The expression level of Akt mRNA in testicular tissue of rats in the high-dose 3 mg/kg/day Cd group was increased (p < 0.05). Our data suggest that Cd affected testosterone levels, and apoptosis was observed in spermatids.

Long-term inhalation of carbon black nanoparticles (CBNPs) leads to pulmonary inflammatory diseases. Histone deacetylase 6 (HDAC6) has been identified as an important regulator in the development of inflammatory disorders. However, the direct involvement of HDAC6 in CBNPs-induced pulmonary inflammatory responses remains unclear. To explore whether HDAC6 participates in CBNPs-induced pulmonary inflammation, human bronchial epithelial cell line (16HBE cells) was transfected with HDAC6 small interference RNA (siRNA) and then exposed to CBNPs at concentrations of 0, 25, and 50 µg/ml for 24 h. Intracellular HDAC6 and intraflagellar transport protein 88 (IFT88) mRNA and protein were determined by real-time polymerase chain reaction and Western blot, respectively. The secretions of inflammatory cytokines including interleukin (IL)-8, tumor necrosis factor (TNF)-α, IL-6, and IL-1β were measured by enzyme-linked immunosorbent assay. CBNPs induced a significant increase in the expressions of IL-8 and IL-6, accompanied by a high level of intracellular HDAC6 mRNA when compared with a blank control group ( p < 0.05). However, there were no significant changes in the levels of TNF-α secretion, intracellular HDAC6 and IFT88 protein induced by CBNPs ( p > 0.05). The HDAC6 mRNA expression was significantly suppressed in HDAC6 siRNA-transfected cells ( p < 0.05). The secretions of IL-8, TNF-α, and IL-6 were significantly less in HDAC6 siRNA-transfected cells than that in normal 16HBE cells with exposure to 25 or 50 µg/ml of CBNPs, but intracellular IFT88 mRNA expression was markedly increased in HDAC6 siRNA-transfected cells when compared with normal 16HBE cells exposed to 50 µg/ml of CBNPs (all p < 0.05). Downregulation of the HDAC6 gene inhibits CBNPs-induced inflammatory responses in bronchial epithelial cells, partially through regulating IFT88 expression. It is suggested that CBNPs may trigger inflammatory responses in bronchial epithelial cells by an HDAC6/IFT88-dependent pathway.

目的 探讨STAT3/p53在邻苯二甲酸二(2-乙基)己酯(di-2-ethylhexyl phthalate,DEHP)诱导大鼠睾丸损伤中的作用.方法 将24只6周龄SD雄性大鼠随机分为对照(玉米油)组及50、100、200 mg/mlDEHP染毒组,每组6只.每日早晨9点,采用灌胃方式进行染毒,每天1次,连续28 d.检测睾丸组织中丙二醛(MDA)含量、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力和细胞凋亡水平及Bax、Bcl-2、细胞色素C(Cyt C)、caspase-9、caspase-3的mRNA表达水平以及p53、p-STAT3蛋白的表达水平.结果 与对照组相比,各剂量DEHP染毒组大鼠睾丸组织GSH-Px活力及500、1 000 mg/kg DEHP染毒组大鼠SOD活力均降低,差异均有统计学意义(P＜0.05);而各剂量DEHP染毒组大鼠睾丸组织MDA的含量均无明显改变;且随着DEHP暴露剂量的升高,大鼠睾丸组织MDA含量呈上升趋势,而SOD、GSH-Px活力均呈下降趋势.与对照组相比,500、1 000 mg/kg DEHP染毒组大鼠睾丸细胞的凋亡指数较高,差异有统计学意义(P＜0.05);且随着DEHP暴露剂量的升高,大鼠睾丸的细胞凋亡指数呈上升趋势.与对照组相比,500、1 000 mg/kg DEHP染毒组大鼠睾丸组织Bax、Cyt C、caspase-9、caspase-3 mRNA的表达水平及250 mg/kg DEHP染毒组大鼠睾丸组织Bcl-2 mRNA的表达水平均较高,而1 000 mg/kg DEHP染毒组大鼠睾丸组织Bcl-2 mRNA的表达水平较低,差异均有统计学意义(P＜0.05);且随着DEHP暴露剂量的升高,大鼠睾丸组织Bax、Cyt C、caspase-9、caspase-3 mRNA的表达水平均呈上升趋势,而Bcl-2 mRNA的表达水平呈下降趋势.与对照组相比,500、1 000 mg/kg DEHP染毒组大鼠睾丸组织p53蛋白的表达水平均较高,而各剂量DEHP染毒组大鼠睾丸组织p-STAT3蛋白的表达水平均较低,差异均有统计学意义(P＜0.05);且随着DEHP暴露剂量的升高,大鼠睾丸组织p53蛋白的表达水平呈上升趋势,而p-STAT3蛋白的表达水平呈下降趋势.结论 DEHP可诱导大鼠睾丸组织发生氧化应激,影响STAT3-p53调控的线粒体途径,诱导大鼠睾丸细胞凋亡,引起睾丸损伤.

目的 探索邻苯二甲酸二(2-乙基己基)酯(di-2-ethylhexyl phthalate,DEHP)和镉联合暴露对青春期雄性大鼠的生殖毒性.方法 采用3×3析因设计,将54只健康6周龄清洁级雄性SD大鼠按体重随机分为9组,分别为对照(0 mg/kg DEHP+0 mg/kg氯化镉)组和250、1000mg/kgDEHP染毒组及1、3 mg/kg氯化镉染毒组以及DEHP(250、1000 mg/kg)+氯化镉(1、3 mg/kg)染毒组,每组6只.采用灌胃方式进行染毒,染毒容量均各为5 ml/kg,连续染毒28 d.收集附睾尾部精子,采用CASA精子分析系统记录精子活力及密度;采用TUNEL法检测睾丸细胞的凋亡情况.结果 与对照组相比,仅1000 mg/kg DEHP+3 mg/kg氯化镉染毒组大鼠的精子活力降低,250 mg/kg DEHP+3 mg/kg氯化镉联合染毒组以及1000mg/kg DEHP+1、3 mg/kg氯化镉联合染毒组大鼠的精子密度和活动精子密度也降低,差异均有统计学意义(P＜0.05).与对照组相比,各染毒组的睾丸细胞凋亡指数均增高,除250 mg/kg DEHP染毒组外,差异均有统计学意义(P＜0.05);联合染毒组的睾丸细胞凋亡指数均低于相同浓度的DEHP染毒组,差异有统计学意义(P＜0.05);250、1000 mg/kg DEHP+3 mg/kg氯化镉染毒组的睾丸细胞凋亡指数均低于3 mg/kg氯化镉染毒组,差异有统计学意义(P＜0.05).250、1000 mg/kg DEHP+3 mg/kg氯化镉染毒对大鼠睾丸细胞凋亡率的影响有交互作用(P＜0.05),表现为为拮抗作用.结论 DEHP和镉联合染毒可以造成青春期雄性大鼠睾丸损伤,其具体作用机制有待进一步研究.

Di (2-ethylhexyl) phthalate (DEHP) is a known environmental endocrine disruptor that impairs development of testis and spermatogenesis. This study aims to explore the effects of STAT3/p53 and PI3K-Akt-mTOR signaling pathway on DEHP-induced reproductive toxicity in pubertal male rat. 24 6-week-old male Sprague-Dawley rats were randomly divided into 4 groups (Control, low-dose, middle-dose and high-dose group) and were treated with increasing concentration of DEHP (0, 250, 500, 1000 mg/kg/day) respectively for 28 consecutive days by intragastric administration. Our results showed that DEHP exposure induced obvious morphological changes of testis, decreased organ coefficient of testis and sperm count, and increased testicular cell apoptosis in the 500 and 1000 mg/kg/day DEHP groups (p < .05). The serum testosterone decreased in a dose-dependent manner after treatment with DEHP. Furthermore, the exposure of DEHP elevated the levels of oxidative stress accompanied by upregulated expression of p53 and reduced expression of STAT3. In addition, compared with the control group, the expression of PI3K, p-Akt and p-mTOR proteins significantly decreased, whereas the downstream autophagy-related proteins phosphorylated ULK1, Beclin-1, Atg7, LC3-II obviously increased in the 250 mg/kg/day DEHP group (p < .05). The expression of p62 was reduced in DEHP-treated groups. Our data indicated that autophagy could be activated to protect testes from DEHP-induced reproductive damage by inhibiting PI3K-Akt-mTOR signaling pathway in the 250 mg/kg/day DEHP group. STAT3/p53-mediated mitochondrial apoptosis pathway might play a major role to cause testis injury and reproductive dysfunction in the 500 and 1000 mg/kg/day DEHP groups.

目的 研究双酚A(bisphenol A,BPA)与壬基酚(nonylphenol,NP)联合作用对子代大鼠睾丸组织氧化应激的影响.方法 采用2×2析因设计,将48只清洁级21日龄SD大鼠随机分为4组,分别为对照组、NP(5 mg/kg)染毒组、BPA(0.5 mg/kg)染毒组和NP(5 mg/kg)+BPA(0.5 mg/kg)染毒组,每组12只(雄雌比为1:3).采用经口灌胃方式染毒至大鼠55日龄,妊娠期和哺乳期(出生21 d)仅对母鼠染毒,隔天一次.每组随机选取6只雄性仔鼠,饲养至60日龄,测定睾丸组织中丙二醛(MDA)含量和总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力.结果 与对照组相比,BPA染毒组仔鼠睾丸组织GSH-Px、T-SOD的活力均较低;NP+BPA染毒组T-SOD活力降低,而MDA含量升高,差异均有统计学意义(P<0.05);而NP染毒组仔鼠睾丸组织GSH-Px、T-SOD的活力及MDA含量均无明显改变.与NP+BPA染毒组比较,NP染毒组睾丸组织T-SOD活力较高,而MDA含量降低;BPA染毒组睾丸组织GSH-Px活力较高,而MDA含量降低,差异均有统计学意义(P<0.05).析因分析结果显示,NP和BPA联合染毒对子鼠睾丸组织GSH-Px活力和MDA含量的影响存在交互作用(P<0.05),分别表现为拮抗效应和协同效应;而对T-SOD活力的影响不存在交互作用(P>0.05).结论 NP和BPA联合染毒可造成雄性子代大鼠睾丸组织氧化损伤,其具体作用机制有待进一步研究.

Purpose: Impaired mucociliary clearance is an initial characteristic of recurrent cough, respiratory infection and chronic respiratory diseases. It has been demonstrated that prolonged inhalation of respirable silica particles results in a variety of pulmonary diseases, but whether the mucociliary system is involved in this process is unclear. This study aims to evaluate the effects of silica particles on mucociliary structure and MUC5B production in respiratory tract.Materials and Methods: C57BL/6 mice were administered with 2.5 mg silica particles through a single intratracheal instillation. The changes of mucociliary structure and MUC5B expression in trachea was evaluated by HE and AB-PAS staining, transmission electron microscopy and immunohistochemistry on days 1, 7, 28 and 84 post-exposure.Results: The mucociliary structure of airway epithelium was obviously impaired by silica particles, showing disordered, shortened or partially lost cilia on the surface, increased mucus in mucous layer and submucosal glands from day 7 to day 84. A variety of ultrastructural abnormalities were discovered in silica-exposed airway cilia, including absence of central pair microtubules, disorganized microtubules and clusters of axoneme on day 1 and 7. The numbers of ciliary axonemes and basal bodies in ciliated epithelial cells were significantly decreased, whereas the proportion of abnormal axonemes was gradually increased with exposure to silica particles (P < 0.05). In addition, silica particles significantly decreased MUC5B expression on the surface of airway epithelium on day 28 and 84, but obviously increased its production in submucosal glands from day 1 to day 84 (P < 0.01).Conclusions: Silica particles could lead to ultrastructural defects in airway cilia, mucus hypersecretion and altered MUC5B expression in trachea, indicating that impaired mucociliary structure and altered MUC5B production might participate in the development of silica-related respiratory diseases.

Prolonged exposure to crystalline silica leads to persistent pulmonary inflammation and progressive fibrosis. Connective tissue growth factor (CTGF) has emerged as a potent proinflammatory and profibrotic regulator to participate in a variety of chronic inflammatory diseases. However, the role of CTGF in silica-induced pulmonary inflammation remains poorly understood. To explore the effect of CTGF on inflammatory responses caused by silica particles, human bronchial epithelial cells (16HBE) were transfected with CTGF siRNA and exposed to silica particles at concentrations of 0, 12.5, 25, 50, 100 μg/ml for 48 h. Intracellular CTGF mRNA and protein expressions were determined by RT-PCR and Western blotting, respectively. The levels of inflammatory cytokines including IL-8, TNF-α, IL-6, IL-1β, IL-17A and TGF-β1 were measured by ELISA kits. Silica particles induce significantly elevated intracellular CTGF mRNA expression in 16HBE cells in a dose-dependent manner when compared with blank control group (P < 0.05). The secretions of IL-8, TNF-α, IL-6 and IL-17A were also significantly increased by silica particles (P < 0.05). After exposure to 25 or 50 μg/ml silica particles, the expression of intracellular CTGF mRNA was significantly inhibited in 16HBE cells when transfected with CTGF siRNA (P < 0.05). The secreted levels of IL-8, TNF-α, IL-6 and IL-17A induced by silica particles were also significantly lower from CTGF siRNA-transfected cells than that from normal 16HBE cells (P < 0.05). Inhibition of CTGF gene attenuates silica-induced inflammatory responses in bronchial epithelial cells, suggesting that CTGF could be a pivotal regulator in the development of silica-induced inflammation.

黑碳(black carbon,BC)又称环境烟尘(soot),是由含碳有机物不完全燃烧而产生的复合型颗粒物,是重要的大气污染物之一;而炭黑(carbon black,CB)是一类几乎只含有元素碳(elemental carbon,EC)的商品化颗粒状产品.多数流行病学研究表明,长期吸入这两种颗粒物均可引起气管、支气管及肺部等组织发生病理改变,进而导致慢性支气管炎、哮喘、慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)、肺癌等多种呼吸系统疾病的发生或发展.但两者在来源、理化特性、暴露人群及对呼吸系统损伤的程度和机制等方面存在较大差异.该文围绕两者的异同点进行综述,有助于更全面了解这两种含碳颗粒物对呼吸系统损伤的特征,为探讨大气颗粒物对呼吸系统的影响及其机制提供有利的科学依据.

目的 研究邻苯二甲酸单-2-乙基己基酯(Mono(2-ethylhexyl)phthalate,MEHP)对小鼠生精细胞(GC-2 spd细胞)凋亡率及Bcl-2 mRNA表达的影响.方法 培养GC-2 spd细胞,用二甲基亚砜(DMSO)溶解MEHP,用不同浓度的MEHP溶液(0、1、10、100、200 μmol/L)对细胞进行24 h染毒.采用MTT法测定细胞活性,荧光定量PCR检测Bcl-2 mRNA的表达情况,流式细胞法测定细胞的凋亡情况.结果 随着MEHP染毒浓度的增加,GC-2 spd细胞活力逐渐降低,细胞凋亡逐渐上升,而Bcl-2 mRNA的表达水平则下降.结论 MEHP可能是通过影响Bcl-2介导的线粒体途径诱导生精细胞凋亡,从而影响雄性生殖功能.

Di(2-ethylhexyl)phthalate (DEHP) is a typical endocrine-disrupting chemical and reproductive toxicant. Although previous studies have attempted to describe the mechanism by which DEHP exposure results in reproductive dysfunction, few studies focused on puberty, a critical period of reproductive development, and the increased susceptibility to injury in adolescents. To elucidate the mechanism underpinning the testicular effects of DEHP in puberty, we sought to investigate the JAZF1/TR4 pathway in the testes of pubertal rats. Specifically, we focused on the role of the JAZF1/TR4 pathway in male reproduction, including the genes JAZF1, TR4, Sperm 1, and Cyclin A1. In the present study, rats were exposed to increasing concentrations of DEHP (0, 250, 500, and 1000 mg/kg/day) by oral gavages for 30 days. Then we assayed testicular zinc and oxidative stress levels. Our results indicated that DEHP exposure could lead to oxidative stress and decrease the contents of testicular zinc. Additionally, significant morphological changes and cell apoptosis were observed in testes exposed to DEHP, as identified by hematoxylin and eosin staining and the terminal deoxynucleotidyl transferase-mediated nick and labeling assay. By measuring the expression levels of the above relevant genes by qPCR, we found the DEHP-induced increased expression of JAZF1 and decreased expression of TR4, Sperm 1, and Cyclin A1. Therefore, we have demonstrated that in vivo exposure to DEHP might induce reproductive toxicity in pubertal male rats through the JAZF1/TR4 pathway and oxidative stress.