Protective effect of Ginkgo biloba extract on testicular tissue of diabetic rats

AIM: To investigate the protective effect of Ginkgo biloba extract (GBE) on diabetic testis and explore its possible mechanism. METHODS: Testicular structure of streptozotocin-induced diabetic rats was observed under light microscopy (LM) and transmission electron microscopy (TEM). Content of malondialdehyde (MDA), NO₂^-/NO₃^- and activity of superoxide dismutase (SOD), nitric oxide synthase (NOS) were determined in testicular homogenate. RESULTS: In diabetic rats, it was manifestated as deformation of seminiferous tubule, atrophy and shedding of germinal epithelium under LM, while expansion of smooth endoplasmic reticulum, formation of fatty vacuoles and decrease of lysosome obviously in the cytoplasm of sertoli cell under TEM, the injury of testicular tissue was improved by GBE. Compared with diabetic rats, activity of SOD increased while activity of tNOS and iNOS, content of MDA and NO₂^-/NO₃^- decreased in GBE-treated rats. CONCLUSION: GBE could effectively prevent the development of diabetic testis and the effect may be partly achieved by resisting lipid peroxidation,restraining the activity of testicular tissue iNOS and reducing the pathological alterations of NO.     

Effect of gingko biloba extract on liver from experimental type 2 diabetic rats

AIM:To study the protective effect of the ginkgo biloba (EGB) extract on liver from experimental type 2 diabetic rats and to explore its possible mechanism. METHODS:Thirty-nine male Sprague-Dawley rats were divided randomly into four groups: normal control group, high-fat group, diabetic group and EGB-treated group. After fed with high-fat diet for 4 weeks, the later two groups were injected with streptozotocin intraperitoneally to induce type 2 diabetes mellitus. EGB-treated group was injected intraperitoneally with EGB at a dose of 8 mg·kg^-1·d^-1, and the other three groups were treated with normal saline of the same volume. After 8 weeks, the morphologic change of hepatic tissue was observed under transmission electron microscope (TEM) and light microscope (LM), respectively. In addition, the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), total nitric oxide synthase (TNOS), inducable nitric oxide synthase (iNOS) and the content of malondialdehyde (MDA), nitric oxide (NO) in liver homogenate were detected biochemically. RESULTS:Obvious liver fatty degeneration, apparent decrease of glycogen granules in cytoplasm of hepatocytes under light microscope and hepatocytes pyknosis, lots of lipid deposits in cytoplasm of hepatocytes, proliferation of hepatic stellate cells and collagen under TEM were observed in diabetic group. The activity of SOD, CAT, GSH-PX decreased but the activity of tNOS, iNOS and the content of MDA, NO^-₂/NO^-₃ increased in diabetic group compared with normal control group. The pathological change was relieved in EGB-treated group. The activity of SOD, CAT, GSH-PX increased, the activity of tNOS, iNOS and the content of MDA, NO^-₂/NO^-₃ decreased in the liver of rats in EGB-treated group compared with diabetic group. CONCLUSION:EGB exerts a beneficial effect on liver in experimental type 2 diabetic rats. Anti-lipid peroxidation and suppression of NO production may be involved in this process.     

Effect of Ginkgo biloba extract on retinopathy in diabetic rats

AIM: To investigate the effect of Ginkgo biloba extract(GBE) on diabetic retinopathy(DR) and its possible mechanism in rats. METHODS: Goto-Kakizaki(GK) rats were used as a DR model, and were treated with different doses of GBE. Normal Wistar rats were used as the control. Blood glucose and retina barrier injury were analyzed, respectively. Ganglion cell apoptosis was detected by TUNEL staining. Moreover, the protein expression of nuclear factor E2-related factor 2(Nrf2), ERK, Bcl-2 and P53, and ERK phosphorylation were examined by Western blot.RESULTS: GBE reduced blood glucose in the DR rats, attenuated retina barrier injury, and decreased the apoptosis of ganglion cells. Furthermore, the expression of Nrf2 and Bcl-2, and phosphorylation of ERK were increased after GBE treatment, whereas P53 expression was decreased. CONCLUSION: GBE protects ganglion cells against apoptosis in DR rats, which may be through activation of Nrf2/ERK pathway and regulating Bcl-2 and P53 expression.     

An enzyme histochemical study of diaphragm in diabetic rats

AIM: The aim of this research is to study the earlier enzyme activity changes of the diaphragm in diabetic rats. METHODS: An enzyme histochemical method was used to observe the changes in the enzyme activities of dehydrogenases,hydrolases and oxidases in 4th week diabetic rat diaphragm. RESULTS: The activites of enzymes including SDH(Succinate dehydrogenase),MDH(Malate dehydrogenase), GDH(Glutamate dehydrogenase), ICDH(Isocitrate dehydrogenase), NADHD(NADH diaphorase), G-6-PD(Glucose-6-phosphate dehydrogenase), ACP(Acid phosphatase) and ANAE(Acid α-naphtyl acid esterase) were increased in diabetic diaphragm compared with the control. LDH (Lactate dehydrogenase)and CCO(Cytochrome oxidase) activities were decreased, whereas NADPHD(NADPH diaphorase) showed no changes in diabetic rats. Eleven kinds of enzyme were analysed with image analysis.Optical density (A) of SDH, MDH, GDH, ICDH, NADHD, G-6-PD, ACP and ANAE in diaphragm of diabetic rats were significantly higher than that of control rats (P<0.01). A value of LDH and CCO in diaphragm from diabetic rats were significantly lower than that of control rats (P<0.01). A value of NADPHD in diaphragm from diabetic rats showed no apparent alteration compared with the control rats(P>0.05). CONCLUSION: Increase in the aerobic capacity, decrease in the glycolytic capacity, and disturbance of lipid and energy metabolism were found in diaphragm of 4th week diabetic rats.     

Ginkgo biloba extract enhances c-jun expression and attenuates motoneuron death induced by root avulsion

AIM: To investigate the influence of Ginkgo biloba extract (EGb761) on c-jun expressions and motoneurons survival following root avulsion. METHODS: One hundred and eighty adult Sprague-Dawley female rats were randomly divided into control and EGb761 groups. Immediately after avulsion of C5-T1 nerve roots, the rats were injected ip with either 1 mL of EGb761 25 mg·kg^-1·d^-1 or the same volume of normal saline, and the treatment repeated everyday. At 4 h to 6 weeks following avulsion, the C7 spinal segments of all rats were collected and prepared for c-jun immunocytochemistry and neutral red stain. The numbers of c-jun positive and survival motoneurons were counted and compared between two groups at each time point. RESULTS: In control rats following avulsion, c-jun positive motoneurons appeared at 4 h, reached its maximum at 1 d and declined to 2 weeks. Avulsion-induced motoneurons death started at 2 weeks, climbed to its maximum at 4 weeks-6 weeks. In EGb761 treated rats, both numbers of c-jun positive and survival motoneurons were more than that in control group at each time point. CONCLUSION: EGb761 attenuates avulsion-induced motoneurons death, and this effect may be related to up-regulation of c-jun gene in avulsed motoneurons.     

Effects of Ginkgo biloba extract on myocardial TGF-β1 and collagen expression and interstitial fibrosis in type I diabetic cardiomyopathy rats

AIM:To study the effects of Ginkgo biloba extract (EGB) on myocardial TGF-β1 and collagen expression and interstitial fibrosis in type I diabetic cardiomyopathy rats. METHODS:Thirty male SD rats were randomly divided into normal control group (CON), diabetes mellitus group (DM) and EGB treatment group (EGB). Streptozocin was intraperitoneally injected into the animals in the latter 2 groups to induce type I diabetic rat model. The rats in EGB group were intraperitoneally injected with EGB. At the end of the 12th week, the body weight of each rat and its left ventri-cular weight, blood glucose, glycosylated hemoglobin and serum insulin concentration were measured. The left ventricular end-diastolic volume (LVEDV), the left ventricular end-systolic volume (LVESV), the left ventricular ejection fraction (LVEF) and the stroke volume (SV) were determined by echocardiography. The content of collagen in left ventricular myocardium, and the expression of transforming growth factor β1 (TGF-β1), procollagen type I and collagen type III were assayed by Sirius red staining, immunohistochemical staining and RT-PCR, respectively. Left ventricular myocardial cells of the neonatal SD rats were isolated and cultured in vitro with low-glucose culture medium (LG group), high-glucose culture medium (HG group) or high-glucose culture medium plus EGB (HG+EGB group). The mRNA levels of TGF-β1, procollagen type I and collagen type III were detected by RT-PCR. RESULTS:Compared with CON group, blood glucose, glycosylated hemoglobin, left ventricular weight index, the content of collagen, and the expression of TGF-β1, procollagen type I and collagen type III in left ventricular myocardial tissues of DM group were significantly increased, while the levels of blood insulin, LVEDV and SV were significantly decreased. However, compared with DM group, blood glucose, glycosylated hemoglobin, left ventricule weight index, the content of collagen, and the expression levels of TGF-β1, procollagen type I and collagen type III in the left ventricular myocardial tissues of EGB-treated rats were significantly decreased, while the levels of blood insulin, LVEDV and SV were significantly increased. Compared with LG group, the mRNA expression levels of TGF-β1, procollagen type I and collagen type III were significantly increased. However, compared with HG group, the mRNA expression levels of TGF-β1, procollagen type I and collagen type III were significantly decreased after treated with EGB. CONCLUSION: EGB retards the process of myocardial fibrosis and improves the cardiac functions in type I diabetic cardiomyopathy rats by down-regulating the expression of TGF-β1, reducing the synthesis and deposition of collagen type I and collagen type III.     

Protective effect of curcumin analogue L6H4 on diaphragm of type 2 diabetic rats

AIM: To investigate the protective effect of curcumin analogue L6H4 on diaphragm of type 2 diabetic rats.METHODS: SPF male Sprague-Dawley rats (n=40) were randomly divided into 5 groups: normal control (NC) group, high fat (HF) group, high fat+L6H4 treatment (FT) group, diabetes mellitus (DM) group and DM+L6H4 treatment (DT) group. The rats in the later 4 groups were fed with high-fat diet. After 4 weeks of high-fat diet fee-ding, the rats in DM and DT groups were intraperitoneally injected with streptozotocin to induce type 2 diabetes melliutus. The rats in FT and DT groups were given L6H4 by gavage for 8 weeks. Blood glucose and blood lipid levels were detected biochemically. Fasting serum insulin (FINS) level was measured by radioimmunoassay and insulin resistance index (HOMA-IR) was calculated. Serum adiponectin (APN) level was measured by ELISA. The morphological changes of the diaphragm were observed under light and transmission electron microscopes. Lipid deposition and the activity of succinate dehydrogenase (SDH) and NADH-tetrazolium reductase (NADH-TR) were observed by enzyme histochemical staining. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in the diaphragm were measured by thiobarbituric acid method and hydroxylamine method, respectively. The protein expression of adiponectin receptor 1 (AdipoR1) in the diaphragm was determined by immunohistochemistry and Western blot. RESULTS: The levels of blood lipids, blood glucose, FINS and HOMA-IR in HF and DM groups were higher than those in NC group, but decreased after L6H4 treatment. The serum APN level in HF and DM groups was lower than that in NC group, but increased after treatment with L6H4. The muscle fibers of the diaphragm were shrunk, fat particles accumulated in the muscle fibers, and the mitochondria were slightly swollen in HF and DM groups. The diaphragmatic fibrosis was obvious in DM group. These lesions were relieved after L6H4 treatment. Compared with NC group, the level of MDA and the activity of SDH and NADH-TR in the diaphragm were increased in HF and DM groups, but decreased after treatment with L6H4. The activity of SOD and the expression of AdipoR1 in the diaphragm were lower than those in NC group, but increased after L6H4 treatment.CONCLUSION: The curcumin analogue L6H4 exerts a protective effect on diaphragm in type 2 diabetic rats. The strengthened protein expression of AdipoR1, the increased serum level of APN, and anti-lipid peroxidation may be involved in the process.     

Study on the variation of trace elements and oxidative stress in diaphragm of the diabetic rats

AIM:To investigate the relationship between trace elements and oxidative stress in diabetic diaphragm. METHODS: Contents of copper(Cu), manganese(Mn), zinc(Zn), iron(Fe),cadmium(Cd),chromium(Cr),cobalt(Co),molybdenum(Mo) and lithium(Li) in the alloxan-induced diabetic rat diaphragm muscles were measured by atomic absorption spectrophotometer, andtheir superoxide dismutase(SOD) activities and malondiadehyde(MDA) contents were detected. RESULTS: The contents of Cu, Zn, Cr and Li were lower and the Fe and Cd were higher in diabetic rat diaphragm muscles than that of the control group, but there were no significant differences in Mn, Co and Mo, between control and diabetic group. SOD activities decreased significantly and MDA contents increased significantly in diaphragm of diabetic rats as compared with the control group.In addition,in diabetic rats diaphragm, there was a positive correlation between Zn and SOD, and negative correlation between Zn and MDA, and Cd and SOD. CONCLUSION: Changes of the contents of Cu, Zn, Fe, Cd, Cr, Li and the increase of oxidative stress in diaphragm were found in the early diabetes, and Zn, Cd play an importment role in oxidative stress of diabetic diaphragm.     

Changes of diaphragmatic function and morphologic features in diabetic rats

AIM:To evaluate the changes of diaphragm contractile properties and morphologic features in experimental diabetic rats.METHODS:Diaphragm contractility was assessed by twitch kinetics,maximal tetanic force (P₀),tension-frequency relationship and fatigue index(FI)in vitro diaphragm strip preparations of 4th week diabetic Wistar rats,and compared with that of the control group.The structure of diaphragm was analysed in diabetic rats and control with Hematoxyline and eosin staining,Heidenhain staining and enzymatic histochemistry.RESULTS:The twitch (Pt),contraction time(CT),half relaxation time(RT_1/2)and FI decreased significantly in diabetic group as compared with the control group.There was no difference in Po between the two groups.The tension of diabetic diaphragm at frequencies of 25,50,75,100 and 125 Hz was significantly decreased compared with that of the normal rats.After diaphragm bundles were subjected to fatiguing stimulations and incubated with aminophylline at 250 mg/L for 30 minutes, the tension of diabetic diaphragm decreased remarkably at both high and low frequencies in comparison with that of the normal rats.In addition,the activity and optical density ofα-phosphoglycerol dehydrogenase in diaphragm from diabetic rats were significantly lower than that from control rats(P<0101).CONCLUSION:The reduction of contractile function,decreased diaphragm fatigue resistance,and damaged structure occurred in 4th week diabetic rat diaphragm.     

银杏叶或银杏叶提取物药理作用机制的研究进展

银杏是我国传统中药,具有重要的药用价值。现对银杏叶或银杏叶提取物在心血管系统、脑及中枢神经系统、抗肿瘤方面的药理作用机制的研究进行了综述。     

银杏幼胚离体培养再生植株的研究

 以佛手银杏幼胚为外植体, 研究了不同大小幼胚在不同培养条件下, 愈伤组织和胚状体的诱导发育情况。结果表明: 大于3 mm的幼胚, 在MK +NAA 1.0 mg·L ^{- 1} +BA 1.0 mg·L ^{- 1}培养基上胚状体诱导率最高, 达到53.6% , 最多的一块愈伤组织形成多达38个胚状体。暗培养不利于胚状体的发生。在MK培养基上添加10%椰汁对胚状体的生长发育有很好的促进作用, 有34.5%生长成苗。     

银杏新品种--恩银1号

‘恩银1号’是从佛手类银杏变种中经单株选择并人工授粉育成的早熟品种，种仁甜糯而香，种实椭圆形，平均10．9g，种核平均3．13g，出仁率高，抗茎腐病。     

Effect of Ginkgo biloba extract on the expression of connective tissue growth factor in the initiating stage of fibrosis in lungs of rats

AIM: To study the effect of Ginkgo biloba extract (GbE) on the expression of connective tissue growth factor (CTGF) in the initiating stage of pulmonary fibrosis of rats after administration of bleomycin (BLM).METHODS: The expression of CTGF in lungs was detected by Western blotting. The content of hydroxyproline was assayed by the method of chloramines T. The content of malondialdehyde (MDA) in plasma was investigated by colorimetry.RESULTS: On day 14 after administration of BLM, the contents of CTGF in lungs and MDA in plasma in BLM+NS group were higher than those in NS group, respectively (P<0.05; P<0.01). On day 30 after BLM, the contents of hydroxyproline in lungs and MDA in plasma in BLM+NS group were higher than those in NS group, respectively (both P<0.01). Treatment with GbE ameliorated the above changes induced by BLM. CONCLUSION: GbE ameliorates the up-regulation of CTGF in the initial stage of fibrosis in lungs of rats after administration of BLM. GbE prevents the hyperoxidative injury in lungs of rats after BLM, which might be one of mechanisms underling the effect of GbE on CTGF.     

银杏苗根生垂乳分泌腔的解剖结构与组织化学研究

以银杏苗木根生垂乳为试材,通过树脂半薄切片技术观察分泌腔的发生发育过程,运用组织化学方法初步研究分泌腔分泌物的主要成分。结果表明：银杏苗根生垂乳中,分泌腔主要分布在皮层中,由一层分泌细胞围绕一个圆形或椭圆形的腔道和2 ~ 3层鞘细胞构成,分泌腔直径平均为566.7 μm。分泌腔由原始细胞团细胞裂溶生而形成,原始细胞团细胞较小,细胞核大,细胞质浓。随分泌腔发育,原始细胞团中央细胞胞间层膨胀、溶解,形成胞间隙,胞间隙扩大到一定程度,中央细胞开始溶解。分泌腔随中央细胞不断溶解而扩大直至分泌细胞停止溶解,分泌腔成熟。根生垂乳顶端皮层及不定芽分生组织中有大量发育程度不同的分泌腔。根生垂乳原始细胞团具有较强的嗜锇性,核蛋白丰富,但淀粉粒较少。中央细胞溶解阶段,分泌细胞及空腔中均有嗜锇物质,空腔中还有大量蛋白质类物质及少量较小的淀粉粒。分泌腔成熟后,分泌腔中基本无蛋白质及淀粉粒,但分泌细胞与鞘细胞均呈现较强的嗜锇性。     

银杏种实生长发育过程中胚乳淀粉体发育观察

对银杏胚乳发育过程进行连续2年的观察结果表明:1)发育成熟的银杏胚乳包括糊粉层与内胚乳2个部分,淀粉体主要积累在内胚乳细胞中,均为单粒淀粉,发育成熟的淀粉体可分为大粒和小粒2种;2)根据淀粉体的形状、大小和发育程度大致可分为淀粉体发生期、淀粉体体积与数量快速增长期、淀粉体发育成熟期3个时期;3)胚乳淀粉体的发生时期及充实部位表现出很大差异,合点端胚乳细胞中的淀粉体发生早,充实快,而珠孔端胚乳细胞中的淀粉体发生迟,充实速度慢;内胚乳外围细胞中淀粉体的充实要高于内部;颈卵器周围的胚乳细胞淀粉体的充实差。     

培养基及培养条件对银杏愈伤组织黄酮产量的影响

在固体培养条件下，研究了不同外植体来源、培养基、抗褐变剂及培养时间等因子对银杏愈伤组织黄酮产量的影响。结果表明：①不同外植体来源的愈伤组织黄酮含量为叶片＞茎段＞子叶；②光照培养的愈伤组织黄酮含量显著高于暗培养愈伤组织黄酮含量；③愈伤组织培养４０～４５ｄ，黄酮含量接近最大值；④Ｗｈｉｔｅ培养基上的愈伤组织黄酮含量显著高于ＭＴ培养基，但黄酮生物产量却相反；⑤不同激素组合中，以ＢＡ与ＮＡＡ组合效果最好；⑥在ＰＡ（植酸）、ＰＶＰ（聚乙烯吡咯烷酮）、ＰＭ（ＰＶＰ＋ＭＥＳ）、ＡＣ（活性炭）和维生素Ｃ等５种抗褐变剂中，以ＰＭ效果最好。黄酮生产最佳培养条件为：叶片愈伤组织在ＭＴ＋ＢＡ１．０ｍｇ／Ｌ＋ＮＡＡ３．０ｍｇ／Ｌ＋糖５％＋ＰＭ（ＰＶＰ２．０％＋ＭＥＳ０．０５％）培养基上光照培养４０～４５ｄ。     

银杏种子繁殖技术研究

以银杏种子为试材,研究了温度、播种基质、赤霉素、氯化钙等因素对银杏种子发芽率的影响。结果表明:恒温、变温处理及播种基质的不同对银杏种子发芽率影响不大;赤霉素和氯化钙处理均能提高银杏种子的发芽率,赤霉素的最佳处理浓度为1 500mg/L,氯化钙的最佳处理浓度为50mg/L。     

银杏实生树与嫁接树光合特性的比较

 对银杏实生树与嫁接树的光合特性及日变化进行了比较研究。结果表明：实生苗嫁接树和二次嫁接树的最大净光合速率、光饱和点和日光合产物积累量均显著高于实生树，这与嫁接提早银杏结果密切相关。嫁接后光合能力的提高与其比叶面积低，单位面积氮含量高有关；但其光合氮利用效率显著低于实生树。实生树与嫁接树光合速率和气孔导度日变化均呈“双峰”型。光合速率的“午休”与强光、高温和低湿度有关，实生树“午休”的原因主要是非气孔限制，两种嫁接树主要是气孔限制。     

银杏摘心处理对枝梢和叶片生长的影响

为探讨银杏夏季摘心对枝叶生长发育的影响,2005年在广西植物研究所银杏良种园内对银杏高接换种树1年生枝梢进行了夏季摘心试验。采用Li-3000A叶面积仪定期测量嫩叶面积;同时在摘心前后对一次梢和二次梢的粗度、长度进行测定,统计二次梢的萌发数量。结果表明,夏季摘心有效地抑制了枝条的加长生长,促进了加粗生长,摘心后一次枝粗是对照的1.52倍;并促进二次枝的萌发和有效叶片数的增加,摘心后枝条上平均每芽着生的叶片数是对照的3.50倍;摘心还促进了叶面积的快速增长,提早叶片成熟时间,有利于光合产物的合成,为提早结果、提高果实产量和品质奠定基础。另外,叶面积与叶柄长和粗之间均呈极显著线性正相关。该项研究将为银杏夏季整形修剪提供理论和实践依据。