博落回提取物猪耐受性评价及其抗结肠炎机制

将健康的50头长大白杂交猪均分为5组,试验Ⅰ、Ⅱ、Ⅲ、Ⅳ组分别按每千克饲料添加博落回散30、150、300、600 mg混饲给药,相当于推荐临床剂量的1、5、10、20倍,对照组(CK)饲料中不添加博落回散,连续饲喂60 d,研究不同剂量博落回散对猪的生长性能、血液学指标、血清生化指标及组织病理学的影响,基于网络药理学探讨博落回提取物(MCE)抗结肠炎的潜在机制。结果表明,与CK相比,饲料中添加30 mg/kg的博落回散可显著提高试验猪的平均日增质量,显著降低料肉比,其他所用剂量的博落回散对试验猪的生长性能均无显著影响;饲料中添加30~600 mg/kg的博落回散的试验猪血液学和血清生化指标均无显著变化,且各脏器组织未见异常病理性损害。可见,饲料中添加30~600 mg/kg博落回散连续饲喂60 d,试验猪具有较好的耐受性。通过SwissTargetPrediction、Genecards、STRING等工具和数据库初步揭示了MCE可能是通过作用于ESR1、BCL2L1、MTOR、MCL1、PTGS2、MAPK8、MMP9、MAPK1、ERBB2和PIK3CA等靶点,并通过ErbB和TNF等信号途径发挥抗炎作用的。     

Safety of standardized Macleaya cordata extract in an eighty‐four‐day dietary study in dairy cows

The objective of this study was to evaluate the safety of a standardized Macleaya cordata Extract Product (MCEP) containing the quaternary benzophenanthridine alkaloids, sanguinarine and chelerythrine, when fed to dairy cows. Thirty‐six dairy cows were randomized into three groups with twelve cows/treatment in two replica pens for each treatment group: control (C) without MCEP added to feed, treatment 1 (SANG‐1000) with MCEP added to feed at 1,000 mg/animal/day (1.5 mg/kg bw/day) and treatment 2 (SANG‐10000) with MCEP added to feed at 10,000 mg/animal/day (15.5 mg MCEP/kg bw/day). After two weeks of acclimation, animals were observed for an 84‐day experimental period, with body weight, feed intake and milk production measured daily. Milk composition was analysed every two weeks. Haematological analyses were performed on Day 0 and Day 84, and clinical chemistry analyses were performed on Day 84 of the study. There was no statistically significant difference (p > .10) among the three groups on body condition score, milk production or milk composition over the study period. There were no significant differences in body weight gain or feed consumption among the three groups. Animals in the SANG‐10000 group had significantly higher mean corpuscular volume (MCV) than the C group (p < .1) and lower mean corpuscular haemoglobin concentration (MCHC) than the SANG‐1000 group (p < .1). Concentrations of sanguinarine and chelerythrine in milk samples collected on Day 84 were below the detection limit (LOD) as measured by high‐performance liquid chromatography‐mass spectrometry (HPLC‐MS/MS). In conclusion, this study presents compelling data supporting the hypothesis that the test product MCEP, when included in the TMR at up to 10,000 mg/animal/day (15.5 mg MCEP/kg bw/day), is well tolerated by dairy cows.     

Pharmacokinetics of sanguinarine,chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half‐life (t_1/2) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (C_max) of DHSA (2.49 ± 1.4 μg/L) was higher that of SA (1.89 ± 0.8 μg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h^?1·kg^?1) with a t_1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.     

Effects of different levels of sanguinarine on antioxidant indices,immunological responses,ileal microbial counts and jejunal morphology of laying hens fed diets with different levels of crude protein

This study was carried out to assess the effects of different levels of sanguinarine on antioxidant indices, immunological responses, serum biochemical parameters, ileal microbial counts and jejunal morphology of laying hens fed on diets with different levels of crude protein (CP). A total of 180 laying hens were subjected into nine dietary treatments with four cages of five birds each. Experimental treatments consisted of three levels of CP (85.0, 92.5 and 100% of Hy‐Line W36 manual recommendation) and three levels of sanguinarine (0.00, 3.75 and 7.50 mg/kg) as a 3 × 3 factorial arrangement of laying hens which fed during a 70‐day feeding trial. The in vitro study showed that sanguinarine exhibited sevenfold and threefold decreased antioxidant activities to inhibit 2‐2‐diphenyl‐1‐picric hydrazyl free radical as well as ferric ion reducing rather than butylated hydroxyl toluene. Although using the decremental levels of CP caused the increase in heterophil‐to‐lymphocyte ratio (p < 0.01), dietary administration of sanguinarine could suppress the serum cholesterol and malondialdehyde concentrations as well as heterophil‐to‐lymphocyte ratio (p < 0.05). Additionally, decreasing CP content resulted in the decreased percentage of albumin (p < 0.05); however, it had no negative effects on humoral immunity. Nonetheless, feeding of at least 3.75 mg/kg sanguinarine led to the remarkable increases in serum gamma globulin concentration (p < 0.01) and secondary (p < 0.05) antibody titres against sheep red blood cells. Moreover, a decline in dietary CP content led to higher villi height and crypt depth (p < 0.05; p < 0.001) and consequently decreased villi height‐to‐crypt depth ratio (p < 0.001) than the optimum level (100% CP). In spite of the effects of sanguinarine on the suppression of Escherichia coli and Salmonella counts (p < 0.05), it markedly enhanced villi height‐to‐crypt depth ratio as well as lamina propria lymphatic follicles extent, simultaneously (p < 0.001). Therefore, in spite of the detrimental effects of feeding low‐CP diets on lymphocytes and serum albumin percentages, and villi height‐to‐crypt depth ratio, the administration of incremental levels of sanguinarine could improve cellular and humoral immunity, decrease ileal microbial counts and in turn improve the intestinal health indices in laying hens.     

血根碱对大鼠离体子宫平滑肌收缩的影响及其机制研究

试验旨在研究血根碱对大鼠离体子宫平滑肌活动的影响。分离8周龄SD雌性未孕大鼠子宫平滑肌,通过BL-420F生物信号采集系统记录子宫平滑肌收缩频率和收缩幅度,计算子宫平滑肌活力。以硫酸阿托品、盐酸普萘洛尔、盐酸苯海拉明及盐酸雷尼替丁为阻断剂,观察血根碱对子宫平滑肌M、β、H₁、H₂受体的关系,探讨血根碱对子宫平滑肌的作用机制。结果显示,血根碱组和元胡止痛片组对缩宫素所致大鼠离体子宫平滑肌收缩频率、收缩幅度和活力均有显著的抑制作用（P<0.05）;应用H₁受体阻断剂苯海拉明和H₂受体阻断剂雷尼替丁后,血根碱对子宫平滑肌抑制作用基本被阻断,而应用M受体阻断剂阿托品,β受体阻断剂普萘洛尔后,血根碱对子宫平滑肌收缩仍具有显著的抑制作用（P<0.05）。综上所述,血根碱对大鼠子宫平滑肌活力具有显著的抑制作用,可能是通过抑制H₁、H₂受体实现,与M、β受体无关。     

高效液相色谱法测定大鼠肝微粒体温孵体系中血根碱的含量

本研究建立了大鼠肝微粒体温孵体系中血根碱的高效液相色谱含量测定方法。采用Shimadzu VP-ODS (150 mm×4.6 mm,5 μm)为色谱柱,以乙腈-0.1%磷酸水溶液(30∶70,V/V)为流动相,以1 mL/min流速进样量10 μL在284 nm波长下进行检测,结果显示500和1000 nmol/L血根碱对照品溶液的方法回收率分别为98.3％和98.9%;250、500、1000 nmol/L标准溶液的日内RSD分别为3.76%、2.88%和3.56%,日间RSD分别为5.19%、4.96%和3.84%;血根碱的摩尔浓度在250~10000 nmol/L内与峰面积呈良好的线性关系,表明该方法简单、快速、灵敏度高、特异性好,可用于血根碱在肝微粒体温孵体系中的含量测定。     

Sanguinarine metabolism and pharmacokinetics study in vitro and in vivo

Sanguinarine (SA) is a benzo[c] phenanthridine alkaloid which has a variety of pharmacological properties. However, very little was known about the pharmacokinetics of SA and its metabolite dihydrosanguinarine (DHSA) in pigs. The purpose of this work was to study the intestinal metabolism of SA in vitro and in vivo. Reductive metabolite DHSA was detected during incubation of SA with intestinal mucosa microsomes, cytosol, and gut flora. After oral (p.o.) administration of SA, the result showed SA might be reduced to DHSA in pig intestine. After i.m. administration, SA and DHSA rapidly increased to reach their peak concentrations (C_max, 30.16 ± 5.85, 5.61 ± 0.73 ng/ml, respectively) at 0.25 hr. Both compounds were completely eliminated from the plasma after 24 hr. After single oral administration, SA and DHSA rapidly increased to reach their C_max (3.41 ± 0.36, 2.41 ± 0.24 ng/ml, respectively) at 2.75 ± 0.27 hr. The half-life (T_1/2) values were 2.33 ± 0.11 hr and 2.20 ± 0.12 hr for SA and DHSA, respectively. After multiple oral administration, the average steady-state concentrations (C_ss) of SA and DHSA were 3.03 ± 0.39 and 1.42 ± 0.20 ng/ml. The accumulation indexes for SA and DHSA were 1.21 and 1.11. The work reported here provides important information on the metabolism sites and pharmacokinetic character of SA. It explains the reasons for low toxicity of SA, which is useful for the evaluation of its performance.     

血根碱安全性试验研究

为观察大鼠连续给予血根碱30 d所产生的毒性反应及停药后恢复情况,为临床应用提供理论依据,将血根碱分别按高、中、低剂量[21、10.5、2.1 mg/(kg·d)]给大鼠连续灌胃给药30d,观察给药期间及停药后14 d大鼠的临床体征、血液常规和生化指标、脏器指数等方面的变化。结果表明,连续给药30 d后与对照组比较,高、中剂量组体重明显增加(P0.05),高剂量组的谷丙转氨酶、谷草转氨酶水平升高(P0.05),其他剂量组的血液生化指标,各剂量组的血液常规、脏器指数均未见显著性变化(P0.05);停药后14 d,高、中剂量组比对照组体重明显增加(P0.05),其他各项检查指标无显著差异性(P0.05)。本研究对血根碱的安全性进行了初步评价,结果可为血根碱的临床应用提供理论依据。