神经生长因子(NGF)对兔角膜缘干细胞增殖的影响

目的:探讨神经生长因子(NGF)对兔角膜缘干细胞增殖作用的影响及最佳有效浓度。方法:取成年大耳白兔角膜缘组织,应用组织块消化培养法进行原代培养,取第2代生长状态良好的干细胞,以5×103个/m l细胞浓度接种于96孔细胞培养板中,依次加入不同浓度的NGF,培养48h后,采用M TT法测定细胞增殖情况。结果:10ng/m l、50ng/m l、100ng/m l、200ng/m l不同浓度组的NGF均有促进角膜缘干细胞的增殖作用,与对照组比较差异均有显著性(P<0.01)。1ng/m l组与对照组、50ng/m l组与200ng/m l组比较差异均无显著性(P>0.05)。结论:NGF可有效促进培养角膜缘干细胞的生长,培养的适宜浓度为10~100 ng/m l,为角膜缘干细胞移植患者术后及角膜外伤或溃疡时局部应用NGF提供了理论依据。     

制作角膜缘干细胞完全缺乏动物模型的新方法

目的提出一种新的角膜缘干细胞完全缺乏模型制作方法,为干细胞缺乏疾病的治疗性研究提供可靠的模型。方法利用9mm环钻划界,板层切除角膜组织,通过临床评分、印迹细胞学、组织病理及免疫组织化学验证模型的成功。结果板层切除术后30~45d,模型眼角膜混浊、上皮缺损、大量新生血管长入,印迹细胞学可见PAS( )的杯状细胞,组织病理学检查上皮符合结膜细胞表型,免疫荧光染色可见结膜杯状细胞特异性标志MUC5AC阳性表达。结论环钻划界加板层切除法为一种可靠、有效的角膜缘干细胞完全缺乏模型制作方法。     

The in vitro pharmacological profile of KR31080, a nonpeptide AT1 receptor antagonist

Summary— KR31080 (2-butyl-5-methyl-6-(1-oxopyridin-2-yl)-3-[[2'-(1H-tetrazol-5-yl) biphenyl-4-yl]methyl]-3H-imidazo[4,5-b] pyridine) is a potent inhibitor of angiotensin type 1 (AT₁) receptors in rabbit aorta and human recombinant AT₁ receptors. In the isolated rabbit thoracic aorta, KR31080 caused a nonparallel shift to the right of the concentration-response curves to angiotensin II (All) with decreased maximal response (pD'₂ = 10.1 ± 0.1), but had no effect on the contractile response induced by norepinephrine. KR31080 inhibited specific [¹²⁵I]AII binding to rabbit aortic membranes (AT, receptors) and [¹²⁵I][Sar¹, Ile⁸]AII binding to human recombinant AT₁ receptors in a concentration-dependent manner with IC₅₀ values of 0.84 ± 0.08 nM and 1.92 ± 0.15 nM, respectively, but did not inhibit specific [¹²⁵I)AII binding to bovine cerebellum membranes (ÀT₂ receptors). In the Scatchard analysis, KR31080 interacted with rabbit aortic AT₁ receptors in a competitive manner, similar to losartan. These results demonstrate that KR31080 is a potent and AT₁ selective angiotensin receptor antagonist which exerts a competitive antagonism in the [¹²⁵I]AII binding assay and insurmountable AT₁ receptor antagonism in the functional study.     

Characteristics of the Nonselective Cation Current （NSCCs） in Rabbit Left Ventricular Epicardial, Midmyocardial and Endocardial Myocytes

Objectives Recent studies have described regional differences in the electrophysiology and pharmacology of ventricular myocardium in canine, feline, rat, guinea pig, and human hearts. This has been shown to be due to a smaller IKs and a lager sodium-calcium exchange current (INa-Ca) and late INa in M region (deep subepicardial to midmyocardial). Studies from our laboratory have found a new repolarization current-nonselective cation current (NSCCs) existing in rabbit right ventricular myocytes. Methods We examined the characteristics of NSCCs in epicardial, M region, and endocardial cells isolated from the rabbit left ventricle with standard microelectrode and whole-cell patch-clamp techniques. The permeability to Na , K , Li , Cs but not to Cl- indicating that it was a nonselective cation current. Gd3 (0.1 mmol/l) and La3 (0.1 mmol/l) can block the current markedly. Results Further characterization of NSCCs was significantly smaller in M cells than in epicardial and endocardial cells. NSCCs current density was significantly smaller in M cells than in epicardial and endocardial cells. With repolarization to -80 mV, INs current density was (-0.44±0.05) PA/PF in endocardial cells, (-0.12±0.05) PA/PF in M cells and (-0.28±0.07) PA/PF in epicardial cells; and with repolarization to 30 mV, INs current density was (1.09±0.29) PA/PF in endocardial cells, (0.38±0.09) PA/PF in M cells and (0.91±0.32) PA/PF in epicardial cells. Conclusions Transmural dispersion of repolarization was due to the heterogeneity of NSCCs in rabbit left ventricle epicardial, endocardial myocytes and M cells. These findings may advance our understanding of the ionic basis for our understanding of factors contributing to the development of cardiac arrhythmias.     

参附注射液对家兔心脏骤停-心肺复苏模型循环恢复的影响

目的观察参附注射液对家兔缺氧型心脏骤停-心肺复苏 (CA- CPR)模型循环恢复的影响.方法 30只家兔随机分为三组,每组 10只;夹闭气管复制缺氧型 CA- CPR模型.预防组 (A组 )夹管前 10min、自主循环恢复后 8、 15min分别静注参附注射液,治疗组 (B组 )自主循环恢复后 8、 15、 22min静注参附注射液,对照组 (C组 )静注生理盐水,时间同 A组;监测夹管前后家兔的心电图、平均动脉压 (PAP)的变化,并记录开始 CPR至自主循环恢复时间和自主呼吸恢复时间、肾上腺素用量、撤呼吸机时间;自主循环维持 4h,并计算 4h存活率.结果三组 CPR成功率、 4h存活率相近;在自主循环恢复时间方面 A组明显快于 C组,在自主呼吸维持时间、肾上腺素用量、撤呼吸机时间方面则两组相近.结论参附注射液可明显缩短家兔缺氧型 CA- CPR模型的自主循环恢复时间,对促进 CA的循环恢复具有积极意义.     

甲地孕酮在家兔和大鼠的药物动力学

用放射免疫法(RIA)测定甲地孕酮(MA)血清浓度并计算了在家兔和大鼠静注或口服的药物动力学参数。MA口服的生物利用度,家兔为64％,大鼠为56％。MA肝微粒体酶代谢实验提示有极性较MA为大的代谢产物形成,~3H-MA十二指肠给药后30min内门静脉血中放射性远高于外周血,也有极性代谢产物形成。     

家兔阴茎皮肤感觉脊神经来源的电生理研究

目的:探讨家兔阴茎感觉神经来源。方法:健康成年雄性新西兰白兔12只,随机均分为两组:每组6只,A组在左侧脊神经上记录,B组在右侧脊神经上记录。对家兔阴茎施加不同大小的机械刺激,用神经单纤维记录技术,在同侧S1~S4脊神经上记录单纤维放电。结果:通过对家兔阴茎施加各种不同机械刺激,在同侧S2~S3脊神经上能记录到放电,S1、S4脊神经上未能记录到放电。左侧脊神经放电纤维数量分别为:S2:39.67±3.14,S3:21.00±2.19;右侧脊神经放电纤维数量分别为:S2:40.00±3.16,S3:19.67±2.58,左右侧差异无显著性(P>0.05)。结论:家兔阴茎皮肤感觉来源于S2~S3脊神经。     

Pharmacological studies on siculine syrup. II: effects on smooth,skeletal and cardiovascular muscle preparations

Earlier pharmacological screening showed that siculine syrup (a traditional herbal remedy purported to be useful in the prevention and treatment of sickle cell pain – crises, due to sickle cell anaemia – SCA) had antisickling and analgesic activities as well as antimicrobial and diuretic effects. SCA is an important haemoglobinopathy in Africa and many other communities/countries worldwide, with relatively high morbidity and mortality. The present study was to determine the effects of the extract on various isolated muscle preparations – smooth, skeletal and cardiovascular. Siculine (4–20 µg/mL), like acetylcholine (40–400 µg/mL), contracted the isolated rat uterus concentration dependently. Similar effects were observed with the guinea‐pig ileum and rabbit jejunum (2–20 µg/mL). In contrast to these effects, the direct (muscle) and indirect (nerve) stimulations of rat phrenic nerve–diaphragm were relaxed by siculine (4 and 8 µg/mL) and d ‐tubocurarine (0.8 µg/mL). Siculine also concentration‐dependently decreased both the rate and force of contraction of guinea‐pig atria and rabbit heart and also resulted in a fall in cat blood pressure in a manner similar to those of acetylcholine. The possible therapeutic and/or toxicological consequences of these effects including the hypotensive activity is noteworthy since siculine syrup is used by the local population for the prevention and treatment of sickle cell pain crises. Copyright © 2008 John Wiley & Sons, Ltd.     

The participation of adenosine receptors in the adenosine 5''-triphosphate-induced relaxation in the isolated rabbit corpus cavernosum penis

AIM: To investigate the participation of adenosine receptors in the adenosine 5'-triphosphate (ATP)-induced relaxation in the corpus cavernosum penis (CCP) of rabbits. METHODS: The ATP-induced relaxation was assessed on the noradrenaline precontracted CCP of rabbits in the presence and absence of 8-(3-chlorostyryl)caffeine (CSC); an adenosine A(2A) receptor antagonist; alloxazine and MRS1754; adenosine A(2B) receptor antagonists; and ARL67156, an inhibitor of ecto-nucleoside triphosphate diphosphohydrolases. RESULTS: Adenosine and ATP relaxed the noradrenaline precontracted CCP of rabbits in a concentration-dependent manner. The adenosine- and ATP-induced relaxations were suppressed by alloxazine and MRS1754, but not by 8-(3-chlorostyryl)caffeine. ARL67156 potentiated the ATP-induced relaxation but not the adenosine-induced one. MRS1754 suppressed the ATP-induced relaxation potentiated by ARL67156. CONCLUSIONS: The above results suggest that, in the CCP of rabbits, the adenosine receptor mediating adenosine-induced relaxation is of the A(2B) receptor and the ATP directly causes relaxation through the A(2B) receptor on the CCP.