1.Hepatic Oxidative Stress and Inflammation in Emphysema and Intermittent Hypoxia Rat Model
Yameng LIU ; Mengli GAO ; Jie CAO ; Jing FENG ; Baoyuan CHEN
Tianjin Medical Journal 2014;(2):148-151
Objective To investigate the effect of emphysema and intermittent hypoxia (IH) on the hepatic oxidative stress and inflammatory injury in rats. Methods Sixty male Wistar rats were randomly assigned to 4 groups, control group (A), emphysema group (B), IH group (C) and emphysema+IH group (D). Group A was normally fed. Group B was exposed to smoke, 30 min per time, twice everyday. Group C was exposed to 5%O2 30 s/Air 90 s for 8 h/d. Group D was exposed to smoke twice, about 30 min each time, and exposed 5%O2 30 s/Air 90 s for 8 h/d. After continues exposure for 8 weeks, five rats in each group were randomly selected for arterial blood gas analysis. The tissue blocks of liver was obtained for pathologi-cal scoring and measurements of liver oxidative stress in the rest 10 rats of each group. HE staining was used to calculate the mean lining interval (MLI) and mean alveolar number (MAN). The hepatic inflammatory factor interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), superoxide dismutase (SOD) activity, catalase (CAT) activity and malondialdehyde (MDA) con-centration were measured in four groups. Results Characteristics of emphysema were found in group B and group D. The values of MLI were significantly higher in Group B and group D than those of group A and group C (P<0.05). The values of MAN were significantly lower in group B and group D than those of group A and group C (P<0.05). The levels of SOD and CAT were significantly lower in group B, group C and group D than those of group A (P<0.05). And the levels of SOD and CAT were significantly lower in group D than those of group B and group C (P<0.05). The values of liver MDA were signifi-cantly higher in group B, group C and group D than those of group A, and the values were significantly higher in group D than those of group B and group C (P<0.05). The liver histological scores and the levels of IL-6 and TNF-αwere signifi-cantly higher in group B, group C and group D than those of group A, and the values were significantly higher in group D than those of group B and group C (P<0.05). Conclusion Emphysema and IH have synergistic action in causing hepatic oxidative stress and inflammation.
2.Effect of intermittent hypoxia on RhoA/ROCK pathway in lung and on the muscularization in pulmonary vascular in rat model
Xiaokun YANG ; Mengli GAO ; Yameng LIU ; Caili LI ; Jie CAO ; Jing FENG
Tianjin Medical Journal 2015;(2):113-116
Objective To explore the effect of intermittent hypoxia (IH) on RhoA/ROCK pathway in lung and on the muscularization in pulmonary vascular in rat model. Methods Wistar rats (n=40) were randomly divided into two groups:the normal oxygen control group (n=20) and the IH group ( n=20). For 4 weeks, rats in control group and IH group were ex?posed to intermittent normal oxygen (21%O2) or IH (5%-21%O2) respectively. Then, mRNA transcription and protein trans?lation levels of RhoA/ROCK were examined by Real-time PCR and Western blot. Expression of proliferation cell nuclear an?tigen (PCNA) andα-smooth muscle actin (SM-α-actin ) of lung and pulmonary artery were detected by immunohistochemis?try. Results RhoA mRNA transcription level(0.463 ± 0.067 vs 0.182 ± 0.040), ROCK mRNA transcription level(0.384 ± 0.062 vs 0.192 ± 0.052), RhoA protein expression level(0.827 ± 0.065 vs 0.424 ± 0.075)and ROCK protein expression level (0.488±0.088 vs 0.336±0.102)were higher in IH group than those in control group(P<0.05);Levels of PCNA in lung tissue [(54.67±1.80)%vs (9.14±0.91)%], PCNA in pulmonary artery [(49.40±1.21)%vs (8.38±1.13)%], SM-α-actin in lung tis?sue [(42.66±1.63)%vs (35.44±1.41)%] and SM-α-actin in pulmonary artery [(62.62±2.53)%vs (45.54±2.58)%] were also higher in IH group than those in control group(P<0.05). Conclusion Rho/ROCK pathway may play an important role in developing pulmonary hypertension (PH) associated with IH;and IH can promote the muscularization in pulmonary vascular to accelerate PH.
3.Effect and mechanism of Kuntai Capsule on premature ovarian failure in rats
Wenjun XU ; Hui GAO ; Yang LI ; Ce LIANG ; Xinpeng LI ; Sa ZHANG ; Yameng CUI
Drug Evaluation Research 2017;40(3):314-318
Objective To explore the effect and mechanism of Kuntai Capsule (KC) on premature ovarian failure in rats.Methods Totally 40 SD female rats were randomly divided into four groups:control group,model group,Kuntai Capsule (KC) group and conjugated estrogens tablets (CET) group.The premature ovarian failure model in rats was made by ig administration of 75 mg/kg tripterygium.After the model was established,rats were continually treated with 0.6 and 0.625 g/kg of KC and CET respectively by ig administration for 36 d.HE staining was used to observe the morphology of ovary and count the number of follicle.The enzyme-linked immunosorbent assay (Elisa) method was used for the detection of follicle stimulating hormone (FSH),luteinizing hormone (LH),serum estradiol (E2),and anti-Mueller tube hormone (AMH) level.The real-time fluorescence quantitative PCR (qRT-PCR) method was used to detect the mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).Results Compared with control group,the number of primordial follicles,preantral follicles and antral follicles were significantly reduced,the number of atretic follicles was increased (P < 0.05),E2 and AMH significantly decreased (P < 0.05),FSH and LH levels increased significantly (P < 0.05),and the levels of VEGF and bFGF mRNA in model group were significantly decreased (P < 0.05).Compared with model group,the number of primordial follicles,preantral follicles,and antral follicles were significantly increased,the number of atretic follicles was significantly reduced (P < 0.05),E2 and AMH significantly increased (P < 0.05),FSH and LH levels significantly decreased (P < 0.05),and the levels of VEGF and bFGF mRNA in KC group were significantly increased (P < 0.05).Conclusion By regulating the level of hormone and up-regulating the expression of VEGF and bFGF,KC can repair the damaged ovarian tissue and promote the growth and development of the follicle,so as to inhibit the premature exhaustion of mRNA.
4.Effects of ionizing radiation on mitochondrial function of mouse hematopoietic stem and progenitor cells
Qi WANG ; Ke ZHAO ; Yameng GAO ; Xin LI ; Yunqiang WU ; Yaxin ZHU ; Zhichun LYU ; Huiying SUN ; Huiying GAO ; Shensi XIANG ; Changyan LI
Chinese Journal of Radiological Medicine and Protection 2022;42(5):321-327
Objective:To study the effect of different doses of 60Co γ-ray ionizing radiation on mitochondrial function in mouse hematopoietic stem and progenitor cells (HSPCs). Methods:C57BL/6 mice were divided into control group, 1 Gy irradiation group and 4.5 Gy irradiation group. The mitochondrial functions were detected at 12 h and 24 h after irradiation, including ROS level, membrane potential, mitochondrial structure, and mitochondrial stress. Bone marrow c-Kit + cells received a single 15 Gy irradiation in vitro, after 24 h, mitochondrial function was detected. Results:It was found that mice leukocytes ( t=12.41, 18.31, 16.48, 14.16, 19.08, 20.25, P<0.05), red blood cells ( t=4.81, 6.62, P<0.05) and platelets ( t=4.33, 6.68, P<0.05) were significantly reduced. The numbers of bone marrow colony formation unit ( t=16.27, 55.66, 17.06, 43.75, P<0.05), and HSPCs ( t=5.16, 11.55, P<0.05) were decreased dose-dependently post-irradiation. Under 1 Gy irradiation, the mitochondrial function and mitochondrial basal metabolic index of HSPCs ( t= 7.36, 3.68, 4.58, 3.15, 3.15, P<0.05) were enhanced at 24 h post-irradiation. Under 4.5 Gy irradiation, mitochondrial number, mitochondrial membrane potential ( t=12.29, 10.46, P<0.05), maximal respiration and spare respiratory capacity were decreased ( t=7.81, 5.78, 6.70, 5.83, P<0.05), ROS level was increased ( t=4.63, 4.12, P<0.05). The basal respiration and oxidative phosphorylated ATP production were reduced at 12 h after irradiation ( t=8.48, 3.80, P<0.05); and the proton leakage was increased ( t=6.57, P<0.05) and coupling efficiency was reduced ( t=11.43, P<0.05) at 24 h after irradiation. In cultured c-Kit + cells, the level of ROS ( t=11.30, P<0.05) and the maximum respiration and spare respiratory capacity were increased ( t=4.25, 3.44, P<0.05) while the mitochondrial membrane potential was decreased ( t=34.92, P<0.05) significantly. Conclusions:A method for systematically assessing mitochondrial function in HSPCs was established, and the effect of ionizing radiation on mitochondrial function of HSPCs was clarified, laying a foundation for further revealing the mechanism of ionizing radiation-induced mitochondrial damage in HSPCs.
5.Comparative study of the radiosensitivity of hematopoietic stem/progenitor cells derived from fetal liver and bone marrow
Yameng GAO ; Ke ZHAO ; Xiongwei ZHAO ; Zhichun LYU ; Siyu LI ; Yunqiang WU ; Huiying SUN ; Huiying GAO ; Shensi XIANG ; Changyan LI
Chinese Journal of Radiological Medicine and Protection 2023;43(8):588-594
Objective:To investigate the difference in the radiation sensitivity of hematopoietic stem and progenitor cells (HSPCs) derived from fetal liver and bone marrow.Methods:HSPCs from fetal liver of 14.5 d embryo or bone marrow of 8 week-old mice were isolated to receive a single dose of 5 or 10 Gy irradiation in vitro using a 60Co irradiator. Twelve hours later, the cell apoptosis, mitochondrial reactive oxygen species (ROS) level, colony formation ability and DNA damage in HSPCs were detected. Freshly isolated HSPCs were injected into lethally irradiated CD45.1 + C57BL/6J mice (4.5 Gy+ 5 Gy with an interval of 30 min) Chimerism rate, lineage constitution, and cell cycle were analyzed 12 weeks after transplantation. Results:Compared with bone marrow HSPCs after irradiation, the percentage of apoptosis in fetal liver HSPCs was significantly higher ( t=16.21, 12.27, P<0.05), the level of ROS was dramatically elevated ( t=68.72, 18.89, P<0.05). At 10 Gy, fetal liver HSPCs could not form colonies at all ( t=12.41, 15.67, 9.46, P<0.05). γ-H2AX immunofluorescence staining showed that the DNA damage of fetal liver HSPCs was more severe after irradiation, and the number of Foci formed was significantly higher than that of bone marrow HSPCs ( t=2.27, 2.03, P< 0.05), which indicated that fetal liver HSPCs were more sensitive to radiation. The chimerism rate of transplanted fetal liver HSPCs was lower than that of bone marrow cells ( t=5.84, P<0.05) with a higher proportion of myeloid lineage, suggesting that fetal liver HSPCs had lower in vivo reconstitution capacity than bone marrow HSPCs and were more prone to myeloid differentiation. The cell cycle of bone marrow HSPCs from transplanted chimeric mice was examined, and the proportion of S-phase was significantly higher in the fetal liver group than that in the bone marrow group ( t=2.89, P<0.05). Mitochondrial stress results showed that fetal liver HSPCs had higher basal respiratory capacity ( t=39.19, P<0.05), proton leakage ( t=6.64, P<0.05), ATP production ( t=9.33, P<0.05), and coupling efficiency ( t=7.10, P<0.05) than bone marrow c-Kit + cells, while respiratory reserve capacity ( t=5.53, P< 0.05) was lower than that of bone marrow c-Kit + cells. Conclusions:HSPCs derived from fetal liver display higher radiosensitivty compared with bone marrow HSPCs, laying the foundation for an in-depth illustration of the effects of radiation on hematopoietic stem cells at different developmental stages.
6.Research Progress in Comprehensive Utilization of Solid By-products in the Process of Industrialization of Chinese Materia Medica
Yingtai TAO ; Qingyi GUO ; Fang CHEN ; Kaiwen TANG ; Yameng LIU ; Xiong LIU ; Jiande GAO
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(3):174-179
With the rapid development and expansion of the scale of the industry of Chinese materia medica,a large number of by-products in the process industrialization of Chinese materia medica have been produced,among which,the solid by-products of Chinese materia medica have been favoured by researchers due to the fact that they are rich in a large number of proteins,cellulose,hemicellulose,lignin,etc.,which can be used in the preparation of high value-added products.Therefore,the authors elaborated on the research on biochemical conversion,thermochemical conversion,resource oriented chemical components,preparation of biomass fuel,new composite materials and high-efficiency adsorbent of solid by-products in the process of industrialization of Chinese materia medica in recent years,aiming to provide theoretical basis for the comprehensive and high-value utilization of the solid by-products of Chinese materia medica and extension of the industrial chain.
7.Preparative optimization and antitumor activity of selenylaed polysaccharides from the extract residue of Sarcandra glabra(Thunb.)Nakai
Cheng HU ; Yameng LIU ; Shijie DAI ; Wei LIU ; Xiangdong GAO
Journal of China Pharmaceutical University 2018;49(1):109-116
Selenylation modification of polysaccharides from the extract residue of Sarcandra glabra(Thunb.) Nakai was operated with sodium selenite as modification agent and barium chloride as the catalyst.The synthesis conditions were optimized by orthogonal experiment with the yields and contents of selenium as indicators.Then, the purity,molecular weight,infrared spectroscopy,thermophysical properties and antitumor activity of the seleny-lated polysaccharides were determined.Results showed that the optimum conditions for preparing selenylated polysaccharides from the extract residue of S.glabra(Thunb.)Nakai were at 70 ℃ for 7 h with 1.0% nitric acid using 1.5 g barium chloride as catalyst.Under the optimum condition,the physicochemical properties of the derivatives were measured and the results showed that the total carbohydrate content was(94.89 ± 0.83)%,the yield was(21.93 ± 0.85)%,the content of selenium was(944.54 ± 13.91)μg/g,the molecular weight was 1 339 kD. In vitro antitumor activity indicated that exposure of HepG2 cells to the increasing concentrations of the crude polysaccharide from the residue of S.glabra(Thunb.)Nakai and its selenylated products decreased cell viability in dose-dependent and time-dependent manner.The selenium polysaccharide demonstrated a better antitumor activity compared with the raw polysaccharide,which could be explored as a potential antitumor drug from abandoned extracted residue for the functional foods and pharmaceutical industries.
8.Comparison of urine proteome in rats intraperitoneally infected by different microorganisms.
Wenshu MENG ; Yameng ZHANG ; Yuanrui HUA ; Yijin JIANG ; Jiawen KONG ; Youhe GAO
Chinese Journal of Biotechnology 2021;37(1):276-289
Different microorganisms can cause intraperitoneal infection. This study was to distinguish different microbial infections by urine analysis. Rats were intraperitoneally injected with Escherichia coli, Staphylococcus aureus, and Candida albicans, separately. Urine samples were collected from rats at 0, 12, 36 and 72 h after infection. Urinary proteins were profiled using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Compared with the control (without infection), a total of 69 differential proteins were identified in rats injected with E. coli. A total of 31 differences proteins were identified in rats injected with S. aureus. A total of 38 differential proteins were identified in rats injected with C. albicans. Urine proteome was different when rats were infected by different microorganisms, suggesting that urine may have the potential for differential diagnosis of different intraperitoneal infections.
Animals
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Chromatography, Liquid
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Escherichia coli
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Proteome
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Rats
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Staphylococcus aureus
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Tandem Mass Spectrometry