MS-222 和丁香酚胁迫对四指马鲅幼鱼鳃及肝脏组织微观结构的影响

【目的】探究不同质量浓度的 MS-222 和丁香酚胁迫对四指马鲅（Eleutheronema tetradactylum）幼鱼鳃和肝 脏组织损伤的影响。【方法】设置 7 个浓度的 MS-222（20、30、35、40、45、50、60 mg/L）和 7 个浓度的丁香 酚（5、10、20、25、30、40、50 mg/L）,分析不同处理对平均全长 4.5（±0.5）cm、平均体质量 1.5（±0.5）g 的四指马鲅幼鱼的静水麻醉效应。【结果】当 MS-222 浓度不小于 35 mg/L 时,麻醉 467 s 后幼鱼开始出现应激反应; 浓度为 50 mg/L 时可将部分幼鱼诱导进入麻醉期,浓度为 60 mg/L 时可在 293 s 后将全部幼鱼诱导进入麻醉期。丁 香酚浓度不小于 25 mg/L 时,麻醉 291 s 后幼鱼开始出现应激反应;浓度为 30 mg/L 时可将部分幼鱼诱导进入麻醉期; 浓度为 40、50 mg/L 时,分别可以在 194 s 和 102 s 将全部幼鱼诱导进入麻醉期。随着 MS-222 和丁香酚浓度的升高, 幼鱼进入同一麻醉时期的时间显著缩短（P ＜ 0.05）,高浓度下四指马鲅幼鱼进入麻醉时期的时间过快,超过耐受 值可能对鱼体产生一定毒害作用,导致鱼体死亡。观察幼鱼组织病理切片发现,随着 MS-222 和丁香酚浓度的升高, 幼鱼肝细胞出现空泡,排列紊乱,核内可见核溶解,肝板结构不清;鳃小片弯曲,末端肿大,严重者可胀破、脱落。【结 论】适度的麻醉剂量可以降低鱼体对外界环境的应激反应,减少耗氧,不同浓度的 MS-222 和丁香酚（5 mg/L 除外） 均可在 3 min 内对四指马鲅幼鱼产生镇静作用,当 2 种麻醉剂浓度过高、麻醉时间过长以及幼鱼长时间处于应激状 态时,会出现死亡现象。     

Berberine promotes apoptosis of human breast cancer cells by regulating miRNA-146a

ATM: To observe the effect of berberine on apoptosis of MCF-7 cells and its potential mechanism. METHODS: The MCF-7 cells were divided into control group and the groups with 3 different doses of berberine. The cell viability was detected by MTT assay, while the cell apoptosis was measured by Hoechst 33258 staining and flow cytometry assay. The protein levels of p-P65, Bax and Bcl-2 were Western blot. The levels of microRNA-146a(miRNA-146a) in the MCF-7 cells were detected by RT-qPCR. The miRNA-146a siRNA was transfected to the MCF-7 cells after an evaluation of transfection efficacy, which was co-incubated with berberine to observe its effects on the mRNA levels of Bax and Bcl-2. RESULTS: Compared with control group, the cell viabilities were decreased significantly in medium and high doses of berberine treatment groups with a dose-dependent manner (P<0.01). The cell apoptosis was increased significantly in medium and high doses of berberine treatment groups dose-dependently (P<0.05). The protein levels of Bax were up-regulated, while those of Bcl-2 and p-P65 were down-regulated significantly by the treatment of berberine (P<0.05). In addition, the miRNA-146a levels were increased significantly in medium and high doses of berberine treatment groups (P<0.05) and showed a dose-dependent manner. The mRNA levels of Bax were decreased, while the mRNA levels of Bcl-2 were increased after transfection with miRNA-146a siRNA and co-incubated with berberine.CONCLUSION: Berberine promotes apoptosis of MCF-7 cells. The mechanism may be related to inhibit the activity of NF-κB by incresing the levels of miRNA-146a.     

Effects of resveratrol on levels of ceramide via regulating miRNA-122 in treating non-alcoholic fatty liver disease

AIM: To observe the therapeutical effects of resveratrol on non-alcoholic fatty liver disease and its potential mechanism. METHODS: Male C57BL/6J mice were fed with high-fat and high-cholesterol diet to established non alcoholic fatty liver disease model, and were administrated with resveratrol at doses of 80 mg/kg and 160 mg/kg. After 4-week treatment, the blood sample was collected for determination of total cholesterol (TC) and triglyceride (TG). The liver tissues were harvested for measuring the liver lipid content. The histopathological examination were conducted with hematoxylin and eosin staining. The ceramide levels in the liver tissues were detected by HPLC-MS. The microRNA (mi-RNA)-122 levels in the liver tissues were detected by real-time PCR. The protein levels of serine palmitoyltransferase (SPT) were determined by Western blot. The HepG2 cells were cultured and divided into 5 groups:control group, model group (induced by 0.25 mmol/L oleic acid), model+resveratrol group (treated with 5 μmol/L resveratrol), miRNA-122 siRNA group and resveratrol+miRNA-122 siRNA group. Except control group, the cells in other groups were stimulated with oleic acid and incubated with respective drugs simultaneously for 24 h. The levels of TC, TG and ceramide in the cells of each group were measured. The protein levels of SPT in each group were determined by Western blot. RESULTS: In non-alcoholic fatty liver disease mice, resveratrol dose-dependently reduced the serum TC and TG levels, decreased the lipid deposition, the ceramide level and the SPT protein level, and increased the level of miRNA-122 in the liver tissues. In the in vitro study, compared with model group, resveratrol reduced the serum TC and TG levels, decreased the ceramide level, reduced the SPT protein level. Compared with control group, the levels of TC, TG and ceramide, and the protein expression of SPT were increased in miRNA-122 siRNA group. Compared with miRNA-122 siRNA group, no statistical difference of TC, TG, ceramide and protein expression of SPT in resveratrol combined miRNA-122 siRNA group was observed. CONCLUSION: Resveratrol significantly reduces lipid accumulation by reduction of miRNA-122 and ceramide levels, and decrease in SPT protein levels in the liver.     

MS-222对杂交鲟幼鱼的麻醉效果及模拟运输试验

为了研究MS-222对杂交鲟的麻醉和运输效果,采用药浴麻醉的方法研究了杂交鲟[西伯利亚鲟(Acipenser baerii)(♀)×史氏鲟(Acipenser schrenckii)(♂)]幼鱼在不同浓度和不同水温下的麻醉效果,同时还模拟进行密封运输试验。结果表明:在17℃、20℃、23℃和26℃水温下,MS-222对杂交鲟幼鱼的有效麻醉浓度分别为30.89 mg/L~42.13 mg/L、31.15 mg/L~45.12 mg/L、38.45 mg/L~54.52 mg/L和49.02 mg/L~60.53 mg/L。随着MS-222溶液浓度升高,杂交鲟幼鱼进入麻醉状态所需时间缩短,其复苏所需时间增加。杂交鲟幼鱼麻醉时间有随水温降低而缩短的趋势,复苏时间有随水温降低而延长的趋势。在22℃±0.5℃水温和40 mg/L浓度下,麻醉运输12 h以1∶8的鱼水比的运输效果最好;麻醉运输24h以1∶15的鱼水比的运输效果最好;麻醉运输48 h以1∶25的鱼水比运输比较合适。试验表明:MS-222对杂交鲟幼鱼有较好的麻醉复苏效果,其适宜浓度为40 mg/L~50 mg/L,在保证较高运输存活率情况下,可降低密封袋的用水量,从而减少运输成本。     

miR-222-3p在鹅肥肝形成中的作用及靶基因验证

为了检测miR-222-3p在填饲鹅不同组织中的表达情况,并对其靶基因进行预测和验证,探讨miR-222-3p在鹅肥肝形成中的功能。运用实时荧光定量 PCR技术检测miR-222-3p 在填饲鹅肝脏、胸肌和腹脂中的表达量;采用生物信息学方法对鹅miR-222-3p的靶基因进行预测,荧光定量PCR技术检测预测靶基因在肝脏中的表达量,并利用双荧光素酶基因报告系统验证其靶向关系。结果显示：相比对照组,miR-222-3p 在填饲鹅肝脏和腹脂中的表达量均显著升高;生物信息学预测结果显示MARF1和B4GALNT3基因在3’UTR区域存在miR-222-3p的潜在结合位点,且这两个基因在鹅肥肝中均显著下调,而双荧光素酶基因报告系统显示只有MARF1基因与鹅 miR-222-3p存在靶向关系。结果表明,miR-222-3p在鹅肥肝中表达量显著上调,且可能通过其靶基因MARF1对鹅肥肝的形成发挥调控作用。     

MS-222对大鳞鲃幼鱼麻醉效果的初步研究

旨在研究MS-222对大鳞鲃幼鱼的麻醉效果。在26±0.5℃的水温条件下,观测了大鳞鲃幼鱼在不同浓度MS-222处理下的麻醉效果,包括麻醉和复苏过程中的行为变化、麻醉过程中呼吸频率的变化以及不同空气暴露时间对麻醉幼鱼复苏时间、复苏率的影响。试验结果表明,大鳞鲃幼鱼的麻醉过程可分为6期,复苏过程分为4期。MS-222对大鳞鲃幼鱼的有效麻醉浓度范围为120~160mg/L。随着麻醉时间及浓度的加大,大鳞鲃幼鱼呼吸频率总体呈下降的趋势。进入5期麻醉的幼鱼,当空气暴露时间在4min以内时,复苏时间随空气暴露时间的增加而减少,当暴露时间在6min以上时,复苏时间随暴露时间的增加而增加,暴露10min以内均能复苏。实验证明,MS-222对大鳞鲃幼鱼有较好的麻醉效果,本实验可为大鳞鲃幼鱼的活体运输以及实验操作等提供理论支持。     

MS-222、丁香酚对多鳞四指马（鲅）幼鱼麻醉效果的研究

本文研究了MS-222和丁香酚两种麻醉剂在不同浓度下对多鳞四指马（鲅）幼鱼的麻醉效果,观测了其不同麻醉阶段的表现,探讨了麻醉时间和温度对其入麻和复苏的影响.结果表明 MS-222、丁香酚分别在55 mg/L、10 mg/L以上才对幼鱼有麻醉效果,最适麻醉剂浓度分别为20±5 mg/L和80±10 mg/L;随着浓度的上升,入麻时间缩短,复苏时间增加.随着温度的上升,入麻时间缩短,复苏时间增加.     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

Anaesthesia of Atlantic cod (Gadus morhua) — Effect of pre-anaesthetic sedation, and importance of body weight, temperature and stress

The efficacy of the anaesthetic agents benzocaine, metacaine (MS-222), metomidate and 2-phenoxyethanol was studied in Atlantic cod (Gadus morhua) with average body weights of 10 ± 4 g, 99 ± 33 g and 1022 ± 274 g at water temperatures of 8 °C and 16 °C. The agents were tested individually and as combination anaesthesia comprising pre-anaesthetic sedation with a low dosage of metomidate or 2-phenoxyethanol followed by anaesthesia with benzocaine or MS-222. All agents were administered through bath immersion with an exposure time of 5 min. The different treatments resulted in average induction and recovery times ranging from 52 ± 6 s to 182 ± 16 s and 77 ± 26 s to 659 ± 46 s respectively. Induction and recovery times varied in relation to water temperature and were generally shorter at 16 °C for all weight groups and treatments compared to 8 °C. For benzocaine and MS-222 induction and recovery times were found to increase with increasing body weight. For metomidate the recovery time increased with increasing weight whereas there were no weight related differences in induction time. No differences in either induction or recovery times associated to body weight were found for 2-phenoxyethanol. Acute stress prior to anaesthesia with MS-222 resulted in significantly shorter induction time and prolonged recovery time, as well as deeper anaesthetised fish. The dosage of MS-222 had to be reduced in order to avoid mortality in fish subjected to acute stress. Combination anaesthesia allowed a reduction of the dosages used for inducing anaesthesia and produced markedly reduced recovery times compared to agents administered individually.