Studies on structural elucidation of Aconitum diterpenoid alkaloid by LC-APCI-MS and effects of Aconitum diterpenoid alkaloid on cutaneous blood flow

The chemical constituents of Aconitum yesoense var. macroyesoense and Aconitum japonicum were examined using high-resolution spectral analysis. Twelve novel alkaloids were isolated from A. yesoense var. macroyesoense together with 20 known alkaloids. Eight novel alkaloids were isolated from A. japonicum together with 15 known alkaloids. An HPLC-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS) method was useful for the simultaneous determination of 21 Aconitum alkaloids found in A. yesoense var. macroyesoense and A. japonicum. These compounds were fairly stable under the conditions used, and the protonated molecules or fragment ions characteristic of the molecule appeared as base peaks in the mass spectra and were used for selected ion monitoring. HPLC-APCI-MS is a very promising approach for structural investigations of positional isomers and stereoisomers. This method was applied successfully to stereoisomeric Aconitum alkaloids differing in configuration at C-1, -6, or -12. Comparison of the APCI spectra showed that the abundance of fragment ions was significantly higher for the C-1, -6, or -12 beta-form alkaloid than for C-1, -6, or -12 alpha-form alkaloid. The main alkaloid constituents in the root of A. yesoense var. macroyesoense, Aconitum alkaloids of the C20-diterpenoid type, kobusine and pseudokobusine, and their acyl derivatives were examined for their peripheral vasoactivities by measuring laser-flowmetrically the cutaneous blood flow in the hind foot of mice after intravenous administration. It is thought that the hydroxyl groups of alkaloids, especially a free OH group of pseudokobusine at C-6, were important for action on the peripheral vasculature leading to dilatation, and the results indicated that esterification of the hydroxyl group at C-15 with either anisoate, veratroate, or p-nitroben-zoate may contribute to enhancement of the activity of the parent alkaloids.     

Laser Doppler measurements of cutaneous blood flow in ageing mice and rats.

Laser Doppler velocimetry (LDV) is a non-invasive technique that measures capillary blood perfusion parameters (blood flow, volume and velocity) and can be used non-invasively to evaluate the effects of ageing on cutaneous blood perfusion. Male Fischer-344 rats (R) of 1, 2, 3, 8, 12, 16, 20 and 24 months and C57BL/6N mice (M) 1, 2, 3, 9, 15, 19 and 22 months (n = 6/mo) were used. Animals were anesthetized with ketamine/xylazine and LDV-measured blood flow was made on the backs of all animals. Five readings for each parameter were recorded in order to obtain a mean value of the individual. Skin biopsies (4 mm) were removed after LDV for assessing epidermal and dermal thickness and number of epidermal cell layers. The number of viable epidermal layers in M remained constant while in R it decreased with age. Epidermal thickness in both M and R decreased from 2-3 months and then remained constant. Dermal thickness decreased from 3 to 22 months in M, and increased in R from 1 to 2 months and remained constant. Blood flow of M increased between 1 and 2 months, remained constant to 19 months, then increased at 22 months; flow in R was constant except at 2 months. Thus age differences in epidermal and dermal thickness, and blood flow of M and R occur and should be considered when evaluating cutaneous toxicity studies in different-aged animals. These changes may potentially alter dermal absorption and/or distribution of xenobiotics.     

地奥心血康对小鼠心肌营养性血流量的影响

采用放射性核素８６Ｒｂ测定小鼠心肌营养性血流量的方法。用地奥心血康８０ｍｇ／ｋｇ，ｉｇ或４０ｍｇ／ｋｇ，ｉｐ，均能增加心肌营养性血流量。＊Ｐ＜０．０１。讨论小鼠心肌对８６Ｒｂ摄取量的大小，在很大程度上取决于心肌内开放毛细血管的数量和面积，在一定条件下，心肌血流量愈大，对８６Ｒｂ的摄取量愈多，所以，心肌对８６Ｒｂ的摄取量可以代表心肌营养血流量。采用８６Ｒｂ测定心肌营养性血流量报道较多，但处理心脏的方法有组织研磨法［２］，组织消化法［３］和直接测定法［１，４］。本文采用直接测定法［１］，操作比较简便。关于注射８６ＲｂＣｌ后处死动物时间的选择，施新献认为心脏对８６Ｒｂ的摄取是一个动态过程，在注射后３０ｓ至１５ｍｉｎ的阶段内，心肌８６Ｒｂ含量高且平稳。因此，选择静注后３０ｓ作为观察心肌对８６Ｒｂ摄取能力的时间标准是比较合适的［５］。地奥心血康４０，８０ｍｇ／ｋｇ，ｉｐ和８０ｍｇ／ｋｇ，ｉｇ，均有不同程度地增加小鼠心肌营养性血流量作用，其中８０ｍｇ／ｋｇ，ｉｐ与双嘧达莫１００ｍｇ／ｋｇ，ｉｐ能显著增加小鼠心肌营养性血流量，其作用强度相似，与生理盐水组比较Ｐ＜０．０１。但地奥心血康２００ｍｇ／ｋｇ１次ｉｇ，反而减少心肌?     

Biphasic response of cutaneous blood flow induced by passive cutaneous anaphylaxis in rats

In the immediate phase of passive cutaneous anaphylaxis, sensitized skin mast cells release various mediators when activated by antigen. The present study investigated the effects of the mediators on cutaneous blood flow at the antigen-antibody reaction site. Induction of passive cutaneous anaphylaxis produced a biphasic response consisting of an initial decrease, followed by a sustained increase, in the cutaneous blood flow. The initial phase was almost eliminated by the 5-hydroxytryptamine receptor antagonist methysergide, whereas the second phase was sensitive to the histamine H(2) receptor antagonist ranitidine. The histamine H(1) receptor antagonist chlorpheniramine, the denervation of sensory nerves with capsaicin, the cyclooxygenase inhibitor indomethacin, or the bradykinin B(2) receptor antagonist D-arginyl-L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl-L-(2alpha,3beta,7abeta)-octahydro-1H-indole-2-carbonyl-L-arginine (HOE140) did not affect the blood-flow changes caused by the anaphylaxis. These results suggest that 5-hydroxytryptamine and histamine H(2) receptors mediate the initial decrease and the subsequent increase in cutaneous blood flow, respectively, induced by passive cutaneous anaphylaxis in rats.     

Effects of the Aconitum alkaloid songorine on synaptic transmission and paired-pulse facilitation of CA1 pyramidal cells in rat hippocampal slices

1. The present study investigated the electrophysiological effects of songorine (1–100 μM), an alkaloid occurring in plants of the Aconitum genus, in rat hippocampal slices.
2. Songorine (10–100 μM) evoked a concentration-dependent increase in the amplitude of the orthodromic population spike and in the slope of the field e.p.s.p. The enhancement was long-lasting and was not reversed by up to 90 min of washout. Songorine failed to affect size and shape of the presynaptic fiber spike which represents the compound action potential of the Schaffer collaterals. This indicates that enhancement of the synaptic response is no consequence of an increased afferent excitability.
3. The antidromically evoked population spike was not affected by songorine at concentrations up to 100 μM suggesting that the enhancement of the orthodromic population spike and of the field e.p.s.p. was not due to an increase in pyramidal cell excitability.
4. The input-output curve for the postsynaptic population spike was shifted to the left implying that a presynaptic fiber spike of the same size elicited a larger postsynaptic response, indicating a decrease in threshold for generation of the population spike.
5. The songorine-evoked increase in excitability was not affected by the NMDA receptor antagonist, D-AP5. However, the effect of songorine was completely abolished by the selective dopamine D₂ receptor antagonist sulpiride (0.1 μM) as well as by haloperidol (10 μM) and was mimicked by application of the dopamine releaser, amantadine (100 mM). In contrast, the selective D₁ receptor antagonist, {"type":"entrez-protein","attrs":{"text":"SCH23390","term_id":"1052733334","term_text":"SCH23390"}}SCH23390, did not block the action of songorine.
6. The results indicate that the plant alkaloid songorine enhances excitatory synaptic transmission which may be due to an agonistic action at D₂ receptors.

     

三七皂苷Rg1对大鼠脑血流量和小鼠微循环的影响及作用机制

目的观察三七皂苷Rg1对脑血流量及微循环的影响,探讨其防治心脑血管疾病,改善预后的作用机制。方法将动物随机分组,分别评价三七皂苷Rg1对大鼠脑血流量,对小鼠脑缺血和小鼠耳廓微循环的影响。结果50 mg.kg-1的三七皂苷Rg1可明显增加大鼠脑血流量,显著延长小鼠断头后喘气时间,且明显增加小鼠耳廓细动脉、细静脉管径及增多耳廓毛细血管数量。结论三七皂苷Rg1可能通过改善动物微循环而增加脑血流量,发挥改善脑缺血的作用。     

Effects of some antihypertensive drugs on cutaneous blood flow and inflammatory skin responses following allergen challenge in guinea pigs

The effects of the antihypertensive drugs clonidine, prazosin, and MK 422 (the active parent diacid of enalapril) were studied on the cutaneous blood flow and allergen evoked inflammatory skin reactions in ovalbumin sensitized guinea pigs. The hypotensive effect of the drugs did not significantly change the basal cutaneous blood flow at the time of allergen challenge. MK 422 (0.02 mg/kg) markedly enhanced the wheal and flare reaction following allergen challenge, whereas clonidine (0.005 and 0.05 mg/kg) inhibited the inflammatory response. Prazosin (0.03 mg/kg) did not significantly influence the wheal and flare reaction. Our results indicate that some antihypertensive drugs (clonidine) could be beneficial to antihypertensive patients with inflammatory diseases, while others (ACE-inhibitors) may enhance their inflammatory disorders.     

黄花乌头茎叶中一个新的Hetisine型生物碱

目的研究黄花乌头茎叶的化学成分。方法采用现代色谱方法进行化学成分分离，并通过各种波谱学分析鉴定化合物的结构。结果从黄花乌头茎叶中分离得到2个化合物，分别鉴定为13-dehydro-1β-acetyl-2α,6β-dihydroxyhetisine (I)和关附庚素(II)。结论I为新化合物。     

Effect of the khat alkaloid (+)norpseudoephedrine on uteroplacental blood flow in the guinea pig

In order to investigate effects of khat chewing on uteroplacental blood flow (+)norpseudoephedrine (NPE) infusions were given to 11 anesthetized guinea pigs in late pregnancy (62-66 days) after unilateral uterine artery ligation at days 30-32. Regional blood flows were determined with radioactive microspheres. Mean arterial blood pressure increased with 25% and heart rate with 9% during NPE infusion. Myoendometrial blood flow was reduced by 31%. Placental vascular resistance (PVR) increased by 56% in the control horn (17 fetuses) and by 82% in the ligated horn (17 fetuses). This vasoconstriction was counteracted by the systemic vasopressor response since placental blood flow remained unchanged. When considering only the 13 growth-retarded fetuses, however, PVR increased by 98% and a 19% reduction of placental blood flow could be demonstrated. These results suggest that the placenta of the growth-retarded fetus may be more sensitive to adrenergic stimulation than the normal placenta. Furthermore, since one of the active constituents of khat, (+)norpseudoephedrine, causes vasoconstriction in the uteroplacental vascular bed it is possible that khat chewing could reduce placental blood flow and, as a consequence, impair fetal growth.     

参麦注射液对小鼠血常规的影响及其脏器保护作用

目的:观察参麦注射液对正常和顺铂化疗小鼠血常规、脏器系数、脏器组织形态学的影响,初步探讨其安全性和化疗辅助疗效。方法:健康小鼠,分为空白对照组、生理盐水对照组(0.125 mL/10 g体重)、参麦注射液组(0.125 mL/10 g体重)、顺铂组(3.5 mg.kg-1体重)、参麦注射液+顺铂组,腹腔注射给予药物7 d后,取血处死,测定血常规;取内脏,肉眼观察,计算脏器系数并切片,做HE染色,观察内脏的受损程度。结果:与生理盐水组相比,参麦注射液组白细胞数显著提高(P<0.01),红细胞和血小板数量有增加趋势,脾脏指数显著上升,心、肝、肺、肾脏器系数无显著变化,各脏器均未见病变;与顺铂组相比,参麦注射液+顺铂组小鼠的肝脏脂肪性病变和坏死得到改善,肾小管上皮细胞水变性明显减轻。结论:研究结果显示参麦注射液能促进正常小鼠的骨髓造血功能,未对小鼠脏器造成毒性损害;并对顺铂所致的小鼠肾、肝损伤有一定保护作用。     

Cytotoxic and trypanocidal activities of cinchona alkaloid derivatives

A series of 27 cinchona alkaloid derivatives ( 1f–w , 2a–e and 3a–d ) were investigated for their cytotoxic and trypanocidal activities using seven different cancer cell lines (KB, HeLa, MCF‐7, A‐549, Hep‐G2, U‐87 and HL‐60), two normal cell lines (HDF and CHO) and bloodstream forms of Trypanosoma brucei brucei, respectively. Four compounds ( 1u , 1w , 2e and 3d ) were identified with promising cytotoxic activity with 50% growth inhibition (GI₅₀) values below 10 μM. Two ( 2e and 3d ) of the four compounds also exhibited potent anti‐trypanosomal activity with GI₅₀ values of 0.3–0.4 μM. All four active compounds represented derivatives modified at their C‐9 hydroxy group. With respect to anti‐proliferative activity and selectivity, 2e (epi‐N‐quinidyl‐N′‐bis(3,5‐trifluoromethyl)phenylthiourea) proved to be the most promising derivative for both cancer cells and bloodstream forms of T. b. brucei. The cytotoxic activity of compounds 1u , 1w , 2e and 3d was attributed to their ability to induce apoptosis in cancer cells. The results demonstrate the potential of cinchona alkaloid derivatives as novel anti‐cancer and anti‐trypanosome drug candidates.     

提高血流量及透析液流量对血液透析充分性的影响

目的探讨提高血流量及透析液流量对血液透析充分性的影响,以期为临床透析治疗提供参考。方法选择2009年12月～2011年12月本院收治的行稳定透析的尿毒症患者20例。血透透析液流量定于500 mL/min,血流量定于250 mL/min时,行2个月透析治疗后,将血透透析液流量定于800 mL/min,血流量定于300 mL/min。比较提高血流量及透析液流量前后尿素氮清除效率(Kt/V)、尿素下降率(URR)、血清清蛋白(ALB)、血红蛋白(Hb)、血肌酐(SCr)、C反应蛋白(CRP)水平及充分透析患者所占百分率情况。结果血透透析液流量为800 mL/min,血流量为300 mL/min时可提高Kt/V及URR,与500 mL/min透析液流量及250 mL/min血流量比较差异有统计学意义(P<0.05),而ALB、Hb、SCr、CRP水平差异无统计学意义(P>0.05);提高血流量及透析液流量后充分透析患者所占百分率为90%,显著高于提高前(70%,P<0.05)。结论 800 mL/min透析液流量及300 mL/min血流量的维持性血液透析可选择性用于常规透析不充分的患者,值得临床推广使用。     

Effects of S-dobutamine on venous blood return and organ nutrient blood flow.

The selective contractile effects of s-dobutamine were studied in vitro in selected canine arteries and vein preparations; propranolol was included to block potential beta-mediated vasodilation. These in vitro data were expanded by quantifying the in vivo effects of s-dobutamine on venous blood return and redistribution of regional nutrient blood flow (NBF) and non-nutrient blood flow (non-NBF) in anesthetized dogs. In in vitro studies with isolated canine arteries and veins, s-dobutamine exhibited vein-selective constriction. At maximally efficacious concentrations of agonist, contractions of carotid, coronary, and femoral arteries in response to s-dobutamine were only 7, 25 and 45% as great as those elicited by norepinephrine (NE). Similarly, in jugular vein, s-dobutamine-mediated contractions were 55% as great as those obtained in response to NE. Coronary and femoral arteries precontracted with NE were relaxed in a dose-related manner by increasing concentrations of s-dobutamine. Effects of NE and s-dobutamine on venous blood return (VR) were compared in dogs. s-Dobutamine increased VR by 49 +/- 10 ml, whereas NE increased VR by 14 +/- 6 ml during 5-min infusion. s-Dobutamine significantly increased coronary NBF in left ventricular (LV) endocardium from 115 +/- 10 to 194 +/- 13 and 263 +/- 9 ml/min/100 g at doses of 10 and 20 micrograms/kg/min, respectively. In addition, LV epicardium flow was increased from 87 +/- 8 to 189 +/- 15 and 262 +/- 11 ml/min/100 g at 10 and 20 micrograms/kg/min, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)