机械性创伤致急性肺损伤大鼠血清肿瘤坏死因子α的动态变化和机制

目的 探讨血清肿瘤坏死因子α(TNF-α)在机械性创伤致急性肺损伤大鼠中的动态变化和机制.方法 将雄性Wistar大鼠40只完全随机分为伪创伤组和创伤组、伪急性肺损伤组和急性肺损伤组,每组10只,利用Noble-Collip创伤仪制备机械性创伤模型,腹腔注射TNF-α制作急性肺损伤模型.全部大鼠均于造模后6 h腹主动脉采血处死,采用ELISA法测定血清TNF-α含量,肺组织行常规组织病理学检查.结果 创伤组大鼠血清TNF-α含量明显高于伪创伤组[(335±28)mg/L比(177±10)ng/L],差异有统计学意义(P＜0.01).病理结果 显示:创伤组和急性肺损伤组肺组织炎症非常明显,而伪创伤组和伪急性肺损伤组基本正常.结论 机械性创伤大鼠血清中TNF-α含量的升高可能与机械性创伤后急性肺损伤有关.动态观察机械性创伤后血清中TNF-α的变化,有助于病情的评估及预后判断,具有重要的临床意义.

Abstract：

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

The protective effect of hesperidin on cardiac and renal tissue damage in DOCA/Salt hypertensive rats北大核心CSCD

Aim To investigate the protective effect of hesperidin (HES) on cardiorenal damage induced by DOCA/Salt hypertension and the underlying mechanisms. Methods Eighteen male SD rats were randomly divided into normal group (Ctrl), model group (DOCA/Salt), and DOCA/Salt with hesperidin group (DOCA/Salt + HES). HES was administered for four weeks. Blood pressure, serum creatinine and blood urea nitrogen were measured. The pathological changes in heart and kidney were examined by HE, Masson and Sirius red staining. The expression of α-SMA, collagen I and TGF-β were detected by Western blot. The mRNA levels of Nlrp3, TNF-α, IL-1β, IL-6 and NOXs were measured using qRT-PCR. Results Compared with the model group, HES administration significantly attenuated the occurrence of DOCA/Salt hypertension, improved renal function indicators of hypertensive rats, reduced renal and cardiac fibrosis, deduced the expression of α-SMA, collagen I and TGF-β, inhibited the expression of Nlrp3, TNF-α, IL-1β and IL-6, and decreased the expression of NOXs in renal and cardiac tissues. Conclusions HES can delay the occurrence of hypertension and protect against hypertension-induced renal and cardiac tissue damage, which may be related to the reduction of inflammatory reaction and oxidative stress by HES. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of IFN-γ and dexamethasone on TGF-β_1- induced human fetal lung fibroblast-myofibroblast differentiation

AIM: To study whether Smads signaling pathway was involved in human fetal lung fibroblast-myofibroblast differentiation induced by transforming growth factor (TGF)-β1 and the role of interferon (IFN)-γ, dexamethasone (DEX) in the fibroblast-myofibroblast differentiation. METHODS: α-Smooth muscle actin (α-SMA), Smad2/3, and Smad7 protein were assessed by Western blot. Collagen protein was analyzed by measuring hydroxyproline. α-SMA and collagen III mRNA were assessed by RT-PCR. Myofibroblasts morphology and Smad2/3 nuclear translocation were assessed by immunohistochemistry. The overexpression of Smad7, a negative mediator of Smads signaling pathway, was acquired by transfection of Smad7 vector. RESULTS: During fibroblast-myofibroblast differentiation induced by TGF-β1, IFN-γ200 μg/L markedly blocked TGF-β1-induced α-SMA protein expression (P<0.01), collagen protein (P<0.01) and mRNA (P<0.05) expression, and myofibroblasts morphological transformation, but DEX 10μmol/L augmented TGF-β1-induced     

肿瘤坏死因子α与白细胞介素6在糖尿病大鼠骨骼肌中表达的实验研究

目的 探讨糖尿病大鼠骨骼肌中炎性细胞因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6的表达及其与细胞凋亡的关系.方法 将Wistar大鼠16只分为实验组和对照组各8只.实验组饲以8周高脂高糖饮食后腹腔内注射小剂量链脲佐菌素(30 mg/kg).对照组仅注射柠檬酸-柠檬酸钠缓冲液.第13周末处死大鼠.观察TNF-α及IL-6的蛋白表达,检测骨骼肌细胞中caspase 3的表达,并计算二者的相关性.结果 与对照组数据[TND-a、IL-6、caspase 3分别为(0.63±0.35)%、(2.05±0.83)%、(10.02±0.82)%]相比,实验组大鼠骨骼肌中TNF-α与IL-6(3.75±1.38%,5.77±0.74%)的表达水平均升高(P＜0.01),caspase 3[(10.02±0.82)%]表达明显高于(P＜0.01),TNF-α、IL-6与caspase3的表达具有相关性(r=0.713,r=0.692,P＜0.05).结论 糖尿病骨骼肌病变时炎症与凋亡同时发生,并可能共同导致胰岛素抵抗.

Abstract：

Objective To evaluate changes of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)expression in quadriceps femoris muscle of diabetic and control rats, and to explore the corelation among TNF-α, IL-6 and caspase 3. Methods Totally 16 adult (aged 8 weeks) Wistar rats were divided into non-diahetic (n =8)group and diabetic (n = 8) group. Diabetic rats were fed by high lipid and glucose diet. After 8 weeks, they were made by a single injection of streptozotocin (STZ) at a dose of 30 mg/kg and control rats were injected by citrate buffer ( pH 4.5 ) in the same dose. Quantitative immunohistochemistry assay for TNF-α, IL-6 and caspase 3 was applied. The quantitative analysis was used to evaluate the positive rate of TNF-α, IL-6 and caspase 3 expression and correlation analysis was used to evaluate the corelation of them. Results TNF-α, IL-6 protein had higher expression (3.75 ± 1.38,5.77 ± 0.74) in quadriceps femoris muscle of diabetic rats than those in controls (P ＜ 0.01 ).Caspase 3 ( 10.02 ±0.82) had higher expression in muscle of diabetic rats than that in controls (P ＜0.01 ). There was significant correlation among TNF-α, IL-6 and caspase 3 (r=0. 713,r=0. 692,P ＜0.05). Conclusions In muscle of diabetic rats, apoptosis is closely related to TNF-α and IL-6 protein which may induce the insulin resistance.     

Targeting fibroblast activation protein inhibits endothelial-mesenchymal transition by affecting cancer-associated fibroblasts derived exosomes北大核心CSCD

Aim To investigate whether targeted inhibition of fibroblast activation protein (FAP) can inhibit the endothelial-to-mesenchymal transition (EndMT) of vascular endothelial cells by affecting exosomes (Exo) of cancer-associated fibroblasts (CAFs) and explore the underlying mechanisms. Methods Primary CAFs and peri-tumor fibroblasts (PTFs) were obtained from lung cancer and peri-cancer tissues, and CAFs-exo and PTFs-exo were collected from culture medium, respectively. Exosomes from CAFs treated with specific FAP inhibitor (3.3 nmol • L-1 SP13786) for 24 h were named as Anti-FAP-exo. HMEC-1 cells were incubated in equal volumes of RPMI 1640, PTFs-exo, CAFs-exo and anti-FAP-exo respectively and named as control group, PTF group, C AF group and anti-FAP group. The scratch assay, Transwell invasion assay and angiogenesis assay were used to detect the migration ability, invasion ability and angiogenesis ability of HMEC-1 cells. Immunofluorescence, immunohistochemistry and Western blot were used to detect EndMT-associated protein expression. Results The migration ability, invasion ability and angiogenesis ability of HMEC-1 cells of CAF group were significantly higher than those of PTF group, whereas there was no significant difference between that of anti-FAP group and PTF group. HMEC-1 cells of CAF group had higher expression of α-SMA, SM22α, p-Stat3 and Snail, and lower expression of CD31 and VE-cadherin than that of PTF group. In addition, HMEC-1 cells of Anti-FAP group had lower expression of α-SMA, SM22α, pStat3 and Snail, and higher expression of CD31 and VE-cadherin than that of CAF group. Conclusions Specific inhibition of FAP could indirectly inhibit the migration ability, invasion ability and angiogenesis ability of vascular endothelial cells via affecting CAFs-exo and Stat3-snail-EndMT pathway may be the potential mechanism. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

机械性创伤致继发性肺损伤大鼠肺组织中叉头盒转录因子A1表达的变化及意义

目的 研究机械性创伤致继发性肺损伤大鼠肺组织中叉头盒转录因子(Fox)A1表达的变化及意义.方法 将雄性Wistar大鼠40只完全随机分为伪创伤组和创伤组,每组20只.利用Noble-Collip创伤仪制备机械性创伤模型.全部大鼠均于造模后6h腹主动脉采血处死,提取肺组织的总RNA和总蛋白.逆转录聚合酶链反应检测FoxA1 mBNA的表达,蛋白质印迹检测FoxA1蛋白的表达.结果 创伤组FoxA1mRNA表达水平高于伪创伤组(P<0.01),是伪创伤组的9.25倍;FoxA1蛋白表达:伪创伤组(0.59±0.04),创伤组(0.91±0.03),创伤组FoxA1蛋白表达水平高于伪创伤组,差异有统计学意义(P<0.01).结论 FoxA1在机械性创伤致继发性肺损伤中起着重要的作用.

Abstract：

Objective To study the effect of secondary lung injury caused by mechanical trauma on the expression change of Forkhead box protein Al ( FoxAl) at both mRNA and protein levels. Methods Forty male Wistar rats were randomly divided into sham group and trauma group, each group 20, Noble-Collip drum was used to establish mechanical trauma rat model. Trie total RNA and protein of lung tissue was extracted after modeling of abdominal aortic blood sampling time points were killed. The expression of FoxAl mRNA in rats' lung tissue was observed by real-time RT-PCR, the expression of FoxAl protein was detected by Western Blot. Results The expression of FoxAl protein in trauma group was higher than that in sham group( sham group 0.59 ± 0.04, trauma group 0.91 ± 0.03, P < 0.01). Conclusion FoxAl plays an important role in secondary lung injury caused by mechanical trauma.     

α-Hederin inhibits growth of lung cancer A549 cells in vitro and in vivo by decreasing c-Myc and HIF-1α dependent glycolysis

OBJECTIVE α-Hederin is an effective component of the traditional Chinese medicine Pulsatilla chinensis,which has been reported to exert many pharmacological activities. However, the effect of α-hederin on metabolism is still unclear. This study aimed to illuminate the role of α-hederin in glucose metabolism in lung cancer cells and investigate the molecular mechanism of α-hederin. METHODS CCK8 and colony formation assays were employed to assess the anti-proliferative effects induced by α-hederin. Glucose uptake, ATP generation, and reduced lactate production were measured using kits, and an A549 tumor xenograft mouse model of lung cancer was used to assess the in vivo antitumor effect of α-hederin(5, 10 mg·kg~(-1)). Glycolytic-related key enzymes were detected by Western blotting and immunohistochemical staining. RESULTS Cell proliferation was significantly inhibited by α-hederin in a dose-dependent manner and that α-hederin inhibited glucose uptake and ATP generation and reduced lactate production. Furthermore, α-hederin remarkably inhibited hexokinase 2(HK2), glucose transporters 1(GLUT1), pyruvate kinase M2(PKM2), lactate dehydrogenase A(LDHA), monocarboxylate transporter(MCT4), c-Myc, and hypoxia inducible factor-1α(HIF-1α) protein expression. Using inhibitors, we proved that α-hederin inhibits glycolysis by inhibiting glycolytic regulators. Moreover, a tumor xenograft mouse model of lung cancer further confirmed that α-hederin inhibits lung cancer growth via inhibiting glycolysisin vivo. CONCLUSION α-Hederin inhibits the growth of non-small cell lung cancer A549 cells by inhibiting glycolysis.The mechanism of glycolysis inhibition includes α-hederin inhibiting the expression of the glycolytic regulatory factors HIF-1α and c-Myc.     

Effect of α-asarone on ethanol-induced learning and memory impairment in mice and its underlying mechanism

OBJECTIVE To investigate the effect ofα-asarone on ethanol-impaired cognitive ability and explore the underlying mechanism in mice. METHODS A mouse model of impaired learning and memory was created by ethanol(2.0 g · kg~(-1), ig). α-Asarone(7.5, 15 and 30 mg·kg~(-1), ip) was delivered 10 min prior to ethanol administration. After 40 min, the locomotor activity of mice with learning and memory impairment was evaluated by the open field test and the behavioral effect of α-asarone was evaluated using the novel object recognition test.Glutamate(Glu) and γ-aminobutyric acid(GABA) levels in the hippocampus were determined by ELISA, and the proteins expression levels of hippocampal α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid(AMPA) receptor(Glu R2), N-methyl-D-aspartic acid(NMDA)receptor(NMDAR2 B), synaptophysin I(SYNΙ), glutamate transporter type 1(GLT-1) and calcium/calmodulindependent protein kinaseⅡ(Ca MKⅡ) were detected by Western blotting. RESULTS There was no significant difference in the horizontal or vertical locomotor activity between the ethanol and normal groups or the 7.5, 15 and 30 mg·kg~(-1)α-asarone groups[F(5, 48)=0.6536, P>0.05; F(5, 49)=1.995, P>0.05]. The recognition index in the ethanol group was significantly decreased as compared with that in the normal group[F(5, 46) =6.739, P<0.05]and was markedly increased in the α-asarone groups as compared with that in the ethanol group(P<0.05), with the exception of the 7.5 mg · kg~(-1)α-asarone group(P>0.05). The hippocampal Glu: GABA ratio in mice was significantly elevated in the ethanol group as compared with that in the normal group(33.42±0.8972 vs 30.79±0.2102, P<0.05) and significantly lower in the α-asarone groups(31.99±0.4986 vs. 33.42±0.8972; 30.97±0.1757 vs. 33.42±0.8972; 30.83 0.1723 vs. 33.42±0.8972, P<0.05). The expression levels of GluR2, NMDAR2B, pSYNⅠand p Ca MKII were significantly higher in the ethanol group as compared with those in the normal group(P<0.05) and obviously lower in the α-asarone groups(P<0.05), with the exception of GluR2, NMDAR2B and pCaMKⅡ in the 7.5 mg·kg~(-1)α-asarone group(P>0.05).And the expression level of GLT-1 was significantly lower in the ethanol group as compared with that in the normal group(P<0.05) and obviously higher in the α-asarone groups(P<0.05). CONCLUSION Pretreatment with α-asarone significantly improved the learning and memory impairment. A possible underlying mechanism is regulation of the calcium signaling cascade to correct functioning of related proteins, and thus, maintain the level of Glu.     

Hypoglycemic effect of Astragalus polysaccharide and its effect on PTP1B

Aim: To examine the effects ofAstragalus polysaccharide (APS), a component of an aqueous extract ofAstragalus membranaceus roots, on protein tyrosine phosphatase 1B (PTP1B), a negative regulator of insulin-receptor (IR) signal transduction, and its potential role in the amelioration of insulin resistance. Methods: Ten-week-old fat-fed streptozotocin (STZ)-treated rats, an animal model of type Ⅱ diabetes mellitus (TIIDM), were treated with APS (400 mg/kg po) for 5 weeks. Insulin sensitivity was identified by the insulin-tolerance test. Further analyses on the possible changes in insulin signaling occurring in skeletal muscle and liver were performed by immunoprecipitation or Western blotting. PTP1B activity was measured by an assay kit. Results: The diabetic rats responded to APS with a significant decrease in body weight, plasma glucose, and improved insulin sensitivity. The activity and expression of PTP1B were elevated in the skeletal muscle and liver of TIIDM rats. Thus the insulin signaling in target tissues was diminished. APS reduced both PTP1B protein level and activity in the muscle, but not in the liver of TIIDM rats. Insulin-induced tyrosine phosphorylation of the IR β-subunit and insulin receptor substrate-1 (IRS-1) were increased in the muscle, but not in the liver of APS-treated TIIDM rats. There was no change in the activity or expression of PTP1B in APS-treated normal rats, and blood insulin levels did not change in TIIDM rats after treatment with APS. Conclusion: APS enables insulin-sensitizing and hypoglycemic activity at least in part by decreasing the elevated expression and activity of PTP1B in the skeletal muscles of TIIDM rats.     

The therapeutic effect of Balanophora polysaccharide on acetic acid gastric ulcer in rats and its mechanism; [蛇 ? 多糖对大鼠乙酸型胃溃疡的治疗作用 ? 机制]北大核心CSCD

Aim To study the therapeutic effect of Balanophora polysaccharide(BPS)on gastric ulcer(GU)induced by acetic acid in rats and to investigateits mechanisms. Methods Sixty male SD rats were randomly divided into sham-operated group, GU model group, omeprazole positive group(3.6 mg·kg-1), and low, medium and high dose of BPS treatment groups(100, 200 and 400 mg·kg-1). The GU model group was prepared by acetic acid cautery method, and the morphology and pathological changes of ulcers were observed by visual observation combined with HE staining, and the ulcer area and inhibition rate were measured and calculated; superoxide dismutase(SOD)activity, malondialdehyde(MDA)content and glutathione peroxidase(GSH-PX)activity were measured by enzymatic assay; tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)content were detected by ELISA. The expression levels of epidermal growth factor(EGF)and epidermal growth factor receptor(EGFR)were measured by immunohistochemistry staining and Western blot. Results Compared with the sham-operated group, obvious ulcer damage was seen in the model group. Compared with the model group, the BPS-treated group showed a significant reduction in ulcer area, an increase in SOD and GSH-PX activity and EGF and EGFR expression levels, and a significant decrease in MDA, TNF-α and IL-6 content. Conclusions BPS has a therapeutic effect on GU in rats, and its mechanism may be related to the inhibition of oxidative stress, suppression of inflammatory stimuli and promotion of regenerative repair of gastric mucosa. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

甲泼尼龙联合环磷酰胺对博来霉素诱导的肺纤维化大鼠模型炎症反应与免疫细胞活性的影响CSCD

Objective To observe the effect of methylprednisolone (MP) combined with cyclo‑ phosphamide (CTX) on inflammation and immune cell activity in bleomycin (BLM)‑induced pulmonary fibrosis rat model. Methods Forty healthy 6 to 8‑week‑old SD rats were randomly divided into blank control, BLM model, BLM+MP, and BLM+MP+CTX groups, with 10 rats in each group. The rat model of pulmonary fibrosis was prepared by intratracheal infusion of BLM (5 mg/kg, only once). From the 7th day of modeling, MP (3 mg/kg) was injected in rats in the BLM+MP group and MP (3 mg/kg)+CTX (8 mg/kg) was injected via tail vein in rats in the BLM+MP+CTX group, once daily for 21 days. The degree of lung inflammation and fibrosis in rats was detected using HE and Masson staining methods. The numbers of granulocytes and neutrophils in bronchoalveolar lavage fluid (BALF) and blood T cell subsets in rats were detected using flow cytometry. Results On the 7th day of modeling, the external morphology, HE and Masson staining results of rat lung tissue showed that BLM‑induced pulmonary fibrosis model was successfully prepared. On the 28th day of modeling, the lung tissue structure of the BLM group was disordered with obvious collagen deposition, the number of granulocytes and neutrophils in BALF increased significantly, the propor‑ tion of blood T cells, CD4+ T cells, and regulatory T cells (Tregs) decreased, the proportion of CD8+ T cells, and the CD4+/CD8+ T cells ratio decreased significantly (all P<0.05). Compared with the BLM group, the degree of pulmonary fibrosis in the BLM+MP+CTX group was improved significantly, the number of granulo‑ cytes and neutrophils in BALF decreased significantly, the proportion of blood T cells, CD4+ T cells and Tregs cells increased significantly, the proportion of CD8+ T cells decreased, and the ratio of CD4+/CD8+ T cells increased significantly (all P<0.05). The improvement effect in rats of BLM+MP+CTX group was better than that of BLM+MP group, and the difference was statistically significant (P<0.05). Conclusion MP com‑ bined with CTX can reduce the degree of inflammatory reaction in rats with pulmonary fibrosis and improve T cell immune activity. © 2023 Chinese Medical Association. All rights reserved.     

银杏叶提取物对糖尿病大鼠肾脏基质金属蛋白酶2表达的影响

Objective To study the effect of ginkgo biloba extract (GbE) on the expression of matrix metalloproteinase (MMP)-2 in kidneys of diabetic rates and to analyze the protecting mechanism of GbE on diabetic nephropathy. Methods Totally 24 SD rats of diabetic nephropathy induced by streptozotocin were randomly divided into two groups: diabetes group and treatment group with GbE, and 12 rats with placebo therapy were enrolled as the normal control group. Rats were killed after 4 ～ 8 weeks treatment and the expression of MMP-2 and type Ⅳ collagen were studies by immunohistochemistry and RT-PCR. Results In diabetic rats, blood sugar, weight and kidney/body weight ratio were decreased significantly and the excretion of 24 hour urinary protein was increased significantly compared to the diabetes group. But the excretion of 24 hour urinary protein was improved in GbE group ( P ＜0.05 ). The expression level of MMP-2 protein and mRNA was decreased significantly in renal glomduri of diabetic rats ( P ＜ 0.05 ) but showed no changes in GbE treatment group compared to diabetes group( P ＞ 0.05 ). The expression level of type Ⅳ collagen was significantly increased in diabetic rats( P ＜0.05 ) but showed no changes in GbE group( P ＞0.05). Conclusion GbE can improve diabetic nephropathy by inhibiting the expression of MMP-2 in diabetic Rat.     

银杏叶提取物对糖尿病大鼠肾脏基质金属蛋白酶2表达的影响

Objective To study the effect of ginkgo biloba extract (GbE) on the expression of matrix metalloproteinase (MMP)-2 in kidneys of diabetic rates and to analyze the protecting mechanism of GbE on diabetic nephropathy. Methods Totally 24 SD rats of diabetic nephropathy induced by streptozotocin were randomly divided into two groups: diabetes group and treatment group with GbE, and 12 rats with placebo therapy were enrolled as the normal control group. Rats were killed after 4 ～ 8 weeks treatment and the expression of MMP-2 and type Ⅳ collagen were studies by immunohistochemistry and RT-PCR. Results In diabetic rats, blood sugar, weight and kidney/body weight ratio were decreased significantly and the excretion of 24 hour urinary protein was increased significantly compared to the diabetes group. But the excretion of 24 hour urinary protein was improved in GbE group ( P ＜0.05 ). The expression level of MMP-2 protein and mRNA was decreased significantly in renal glomduri of diabetic rats ( P ＜ 0.05 ) but showed no changes in GbE treatment group compared to diabetes group( P ＞ 0.05 ). The expression level of type Ⅳ collagen was significantly increased in diabetic rats( P ＜0.05 ) but showed no changes in GbE group( P ＞0.05). Conclusion GbE can improve diabetic nephropathy by inhibiting the expression of MMP-2 in diabetic Rat.     

Bioavailability,tissue distribution,and excretion characteristics of the novel carbonic anhydrase inhibitor tolsultazolamide in rats

Aim： Tolsultazolamide, a novel carbonic anhydrase inhibitor, is designed for the prophylaxis and treatment of acute mountain sickness. The aim of this study was to investigate the pharmacokinetics, tissue distribution, and excretion characteristics of tolsultazolamide and the sex difference in pharmacokinetics in rats. Methods： For pharmacokinetic study, rats were intravenously injected tolsultazolamide at I and 2 mg/kg or orally administered tol- sultazolamide at 20, 40, or 80 mp=/kg） in a pharmacokinetic study. The concentrations of tolsultazolamide in plasma were determined with high-performance liquid chromatography, with a liquid-liquid extraction. For tissue distribution study, tolsultazolamide （80 mpJkg） was orally administered to overnight fasted rats （six per group and three per sex）. Samples were collected from the brain, heart, lung, liver, spleen, muscle, kidney, stomach, fat, intestines, pancreas and sexual gland. For excretion study, tolsultazolamide （40 mg/kg） was orally administered to 6 rats （three per sex）. The urine, feces, and bile samples were collected at 24, 48, and72 h. Results： After its intravenous administration, tolsultazolamide was rapidly eliminated from the plasma, with T~/2 of about 60-90 min. The AUCo_t and the initial concentration （CO） values were proportional to the intravenous doses. After its oral administration, tolsult- azolamide showed dose-independent pharmacokinetic characteristics, with Tmax and T~/2 of approximately 2 h and 5-7 h, respectively, and good oral absolute bioavailability of about 60%. Tolsultazolamide was distributed widely in various tissues. The highest tolsult- azolamide levels were detected in the stomach, intestine, spleen, lung, and kidney. Total excretion of unchanged tolsultazolamide in the urine, feces, and bile was less than 2%. The Cm,x and AUC of tolsultazolamide were significantly higher in female rats than those in male rats. Clearance and volume of distribution were greater in male rats than those in female rats. The oral absolute bioavailability was also significantly different between female rats （about 83%） and male rats （about 37%）. Conclusion： Tolsultazolamide was well absorbed and widely distributed in the rat, and very little of the unchanged form was excreted. Sex had a significant effect on the pharmacokinetics of tolsultazolamide.     

银杏叶提取物对糖尿病大鼠肾脏基质金属蛋白酶2表达的影响

目的 观察银杏叶提取物(GbE)对雄性SD糖尿病大鼠肾脏基质金属蛋白酶2(MMP-2)和Ⅳ型胶原表达的影响,从而探讨银杏叶对糖尿病肾病的保护机制.方法 24只链脲佐菌素诱导的SD大鼠完全随机分成糖尿病对照组(DM组)和GbE组,并以12只正常雄性SD大鼠作为正常对照组(NC组).分别于4、8周后,观察各组大鼠体重、肾重、随机血糖、尿蛋白排泄率等生化指标,并用免疫组化和逆转录聚合酶链反应方法 观察糖尿病大鼠肾脏MMP-2及Ⅳ型胶原的表达.结果 糖尿病大鼠的随机血糖、体重、肾脏/体重明显下降,尿白蛋白排泄率明显升高[DM组(31.59±4.59)mg/24 h比NC组(8.74±2.08)mg/24 h,P＜0.05],GbE组糖尿病大鼠尿白蛋白排泄率[(21.38±3.86)mg/24 h]较DM组有所下降(P＜0.05).8周时,与正常对照组相比,DM组大鼠肾小球MMP-2蛋白的表达水平明显下降[(0.49±0.07)mg/24 h比(0.15±0.04)mg/24 h,P＜0.05],而GbE组DM组大鼠肾小球MMP-2蛋白的表达水平[(0.48±0.07)mg/24 h]较DM组无明显下降(P＞0.05);DM组大鼠Ⅳ型胶原的表达水平较NC组明显升高[(3.48±0.54)mg/24h比(1.68±0.51)mg/24 h],而GbE组糖尿病大鼠较NC组无明显升高[(2.55±0.50)mg/24 h比(1.68±0.51)mg/24 h].结论 GbE可通过抑制糖尿病大鼠MMP-2过表达,进而对糖尿病肾病发挥保护作用.

Abstract：

Objective To study the effect of ginkgo biloba extract (GbE) on the expression of matrix metalloproteinase (MMP)-2 in kidneys of diabetic rates and to analyze the protecting mechanism of GbE on diabetic nephropathy. Methods Totally 24 SD rats of diabetic nephropathy induced by streptozotocin were randomly divided into two groups: diabetes group and treatment group with GbE, and 12 rats with placebo therapy were enrolled as the normal control group. Rats were killed after 4 ～ 8 weeks treatment and the expression of MMP-2 and type Ⅳ collagen were studies by immunohistochemistry and RT-PCR. Results In diabetic rats, blood sugar, weight and kidney/body weight ratio were decreased significantly and the excretion of 24 hour urinary protein was increased significantly compared to the diabetes group. But the excretion of 24 hour urinary protein was improved in GbE group ( P ＜0.05 ). The expression level of MMP-2 protein and mRNA was decreased significantly in renal glomduri of diabetic rats ( P ＜ 0.05 ) but showed no changes in GbE treatment group compared to diabetes group( P ＞ 0.05 ). The expression level of type Ⅳ collagen was significantly increased in diabetic rats( P ＜0.05 ) but showed no changes in GbE group( P ＞0.05). Conclusion GbE can improve diabetic nephropathy by inhibiting the expression of MMP-2 in diabetic Rat.     

实验性变态反应性脑脊髓炎大鼠L-选择素的表达及意义

Objective To investigate the effect of L-selectin in experimental allergic encephalomyelitis (EAE) of Wistar rats.Methods The rats were randomly divided into four groups;the normal group,the CFA group, the LMS group and the model group;The animal models were established in rats by immunization with myelin basic protein of spinal cord of guinea pig and complete Freund's adjuvant(CFA).The symptom of EAE was observed; pathological feature and myelin of brain and spinal were detected with HE stain and Loyez's stain respectively.The number of positive vessels of L-selectin expression was detected by immunohistochemistry.Results 100% experimental Wistar rats treated with MBP and levamisole developed EAE,but none of the other groups.The number of positive vessels of L-selectin expression was (31.86 ± 1.39) in model group, obviously higher than that of in the normal group (1.38 ±0.18) ,the CFA group( 1.50 ±0.27) and the LMS group(7.25 ±0.59) (all P <0.05) ;The inflammation cells were found around vessels and demyelination were seen in white matter of brain and spinal cords.Conclusion The expression of L-selectin should play an important role in EAE.     

Effect of aqueous extract of Semen Cassiae on endogenous metabolomics in urine of rats北大核心CSCD

OBJECTIVE: To investigate the effect of aqueous extract of Semen Cassiae (AESC) on endogenous metabolites in urine of rats by metabolomics based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) to reveal the possible ways of metabolism and mechanism of action in rats caused by AESC. METHODS: Twsenty-eight male Sprague-Dawley (SD) rats were randomly equally divided into 4 groups: such normal control group, AESC 1.5, 5 and 15 g·kg-1 groups. After intragastric administration for 14 d, the urine was collected with metabolic cages. The urine metabolic profiling was analyzed using UPLC-QTOF-MS, based on which the principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were established for metabolomic analysis. Potential biomarkers were screened using variable importance in the projection (VIP) and t test. RESULTS: The results of PCA showed that samples of each group were clustered, all the groups were separated, and that the distance between AESC groups and normal control group was increased in a dose-dependent manner. The relative content of proline betaine and uric acid were 18.4±2.3 and 15.7±2.0, 16.3±4.5 and 14.7±3.0 in the AESC 5 and 15 g· kg-1 groups, significantly lower than that of the normal control group, which was 25.0±3.4 and 29.0±4.8(P<0.01), but that of AESC 1.5 g · kg-1 group did not statistically differ from that of normal control group. In AESC 1.5, 5 and 15 g·kg-1 groups, the relative content of glycine and taurine was 10.0±1.4 and 8.0±1.4, 3.6±0.7 and 66.5±7.3, 45.8±23.6 and 23.0±9.8, which was significantly lower than that of the normal control group, which was 14.6±1.9 and 102.5±25.8(P<0.01). The relative content of 1,7-dimethylguanosine was 4.5±1.2 and 4.6±0.1 in AESC 1.5 and 15 g·kg-1 groups, significantly lower than that of the normal control group, which was 6.5±0.8(P<0.05), but AESC 5 g·kg-1 group did not statistically differ from normal control group. The relative content of citric acid was 26.6±6.3 in the AESC 15 g·kg-1 group, significantly lower than that of the normal control group, which was 67±14(P< 0.01). The relative content of citric acid was 104+20 in the AESC 1.5 g·kg-1 group, significantly higher than that of the normal control group (P<0.01), but AESC 5g·kg-1 group did not statistically differ from normal control group. CONCLUSION: AESC can remarkably change endogenous metabolites and mainly affect the pathways of taurine,purine, amino acid and energy metabolism.     

长春西汀对慢性阻塞性肺疾病大鼠炎症干预作用的探讨

目的 探讨长春西汀对慢性阻塞性肺疾病(COPD)慢性炎症的作用.方法 将40只雄性Wistar大鼠随机分为正常对照组、COPD模型组、长春西汀5 mg/kg组、2.5 mg/ks组和1.25 mg/ks组,每组8只.采用熏香烟加气管内注入脂多糖法建立COPD大鼠模型.长春西汀组(共3组)从注入脂多糖次日起给予长春西汀腹腔注射,每日1次.观察各组大鼠肺组织病理改变,检测血清中肿瘤坏死因子(TNr)-a、白细胞介素(IL)-8、C反应蛋白(CRP)的浓度以及支气管肺泡灌洗液(BALF)中TNF-a、IL-8的浓度.结果 COPD模型组肺组织病理改变符合人类COPD的病理特点;长春西汀5 mg/kg组和2.5 mg/kg组病理改变较模型组轻;上述两组血清IL-8浓度分别为(18.40±2.40)和(19.30±3.11)ng/L,比COPD模型组[(23.81±3.54)ns/L]低,血清TNF-a浓度分别为(39.34±3.43)和(40.47±3.09)ns/L,比COPD模型组[(46.65±4.42)ns/L低],血清CRP浓度分别为(4.28±0.22)和(4.35±O.26)ms/L,比COPD模型组[(4.69±0.19)ms/L]低;上述2组肺泡灌洗液中IL-8浓度分别为(20.09 ±2.88)和(21.03±2.21)ng/L,低于COPD模型组[(25.02±2.92)ng/L],TNF-a浓度分别为(40.41±4.40)和(41.18±5.33)ng/L,低于COPD模型组[(48.81±4.92)ng/L],差异均有统计学意义.长春西汀1.25 mg/ks组各项指标与COPD模型组比较,差异无统计学意义(P>0.05).结论 长春西汀能降低COPD大鼠血清和BALF中炎性因子的水平,减轻气道及肺组织炎症,对COPD大鼠的炎症反应有一定的抑制作用.

Abstract：

Objective To observe the effect of vinpecetine on inflammatory factors and lung pathology of the rats with chronic obstructive pulmonary disease(COPD),and to investigate the therapeutic action of vinpecetine on COPD.Methods Totally 40 Wistar rats were randomly divided into the normal control group,the COPD model group and three viupocetine treated groups.The COPD rat model wag established by intratracheal instillation of lipepalysaceharide and exposure to cigarette smoke.The three intervention groups were intrapedtonealy injected with vinpecetine respectively at the dose of 1.25 mg/kg,2.5 mg/kg and 5 mg/kg before exposing to cigarette smoke.Pathologic changes of the lung tissue,interlukin(IL)-8 and tumor necrosis factor (TNF)-a levels in bronchial alveolarhvage fluid(BALF)and seruln,and CRP level in the serum were determined.Results The pathological changes in the COPD rat model were coincident with the changes in human.Compared with the COPD model group,the two groups treated with vinpecetine at the dose of 2.5 mg/kg and 5 mg/kg showed a significan t decrease in IL-8[(18.40 ±2.40)ng/L,(19.30±3.11)ng/L respectively vs(23.81±3.54)ng/L],TNF-a[(39.34±3.43)ng/L,(40.47±3.09)ns/L respectivelyvs (46.65±4.42)ng/L],and CRP[(4.28±0.22)mg/L,(4.35±0.26)mg/L respectively vs(4.69±0.19)mg/L]in serum.In the BALF of those the BALF of those two groups, the levers of Ⅱ-8[(20.09 4±2.88)ng/L,(21.03±2.21)ng/L]and TNF-a[(40.41±4.40)ng/L,(41.18 4±5.33)ng/L]were also significantly lower than those in the moder group[(25.02±2.92).g/L,(48.81±4.92)ng/L].The vinpocetine treated group at the dose of 1.25mg/kg ad no significant difference, compared with the COPD model group. Conclusions Vinpocetine can reduce the levels of inflammatory factors in BALF and the serum of the rats with COPD and decrease inflammation of the airway and lung tissue. Accordingly vinpecetine can inhibit the inflammation of the COPD rat model.