自聚焦超声在动物离体皮下脂肪组织中的融脂效应研究

目的：探索高频聚焦超声和低频聚焦超声在不同辐照时间和功率的条件下,对离体动物猪的皮下脂肪不同层产生生物学融脂效应的规律。方法：将离体动物猪脂肪生物实验分为两组,分别使用工作频率为6．5MHz和3MHz的换能器。两组使用的超声功率为2～5w,辐射时间为5S、10S、20S、4s、60s和80S;辐照后进行MTT检测脂肪细胞活性并计算融脂效应的面积。描述离体动物猪的皮下脂肪层生物学融脂效应,讨论不同条件下生物学融脂效应之间的区别。结果：随着超声功率和辐照时间的增加,脂肪组织形成融脂效应的程度呈规律性变化。辐照时间为5s时脂肪组织破坏不明显,辐照时间〉10s时脂肪组织则明显被破坏。相同超声功率和辐射时间条件下聚焦超声在深脂肪层的破坏程度比浅表脂肪层高。3MHz聚焦超声在辐射时间≥40s可穿过约5mm的表皮并凝固性破坏皮下不同层次的脂肪组织。结论：高频和低频聚焦超声均可有效破坏脂肪组织。     

Regulation of sympathetic nerve activity by L-carnosine in mammalian white adipose tissue

Previously, we showed that l-carnosine, a bioactive dipeptide, influences the sympathetic nerve activity innervating kidney and brown adipose tissue. Because the autonomic nervous system plays an important role in the regulation of lipid metabolism, we investigated the in vivo effects of L-carnosine on the sympathetic nerve activity innervating white adipose tissue (SNA-WAT) and lipolysis. We found that intraperitoneal (ip) administration of L-carnosine at doses of 100 ng/rat and 10 microg/rat elevated and suppressed SNA-WAT, respectively. The effect of lower dose of L-carnosine (100 ng/rat) was eliminated by pretreatment with diphenhydramine hydrochloride, a histamine H(1) receptor antagonist. In contrast, the effect of higher dose of L-carnosine (10 microg/rat) was suppressed by thioperamide maleate salt, a histamine H(3) receptor antagonist. Moreover, ip administration of 100 ng and 10 microg of L-carnosine increased and decreased the levels of plasma free fatty acids (FFAs), respectively. The changes of plasma FFAs resulting from the exposure to 100 ng and 10 microg of L-carnosine were diminished by the beta-adrenergic receptor blocker propranolol hydrochloride and the muscarinic receptor blocker atropine sulfate, respectively; and eliminated by the corresponding histamine receptor antagonists, which eliminated the changes in SNA-WAT. Our results suggest that low doses of L-carnosine may regulate the lipolytic processes in adipose tissue through facilitation of the sympathetic nervous system, which is driven by histamine neurons through the H(1) receptor, and that the beta(3)-receptor may be involved in this enhanced lipolytic response. High doses of L-carnosine, on the other hand, may lower lipolysis by suppressing sympathetic nerve activity via the H(3) receptor, and the muscarinic receptor may be related to this response.     

沉默PLIN1基因与异丙肾上腺素对3T3-L1脂肪细胞脂解的机制探究

目的 研究沉默PLIN1基因与异丙肾上腺素(ISO)联合作用对3T3-L1脂肪细胞脂解的影响及机制探讨。方法 sh-PLIN1干扰载体成功转染后,以浓度为10μM的ISO处理3T3-L1脂肪细胞。采用油红O进行脂滴染色,观察脂滴形态;采用酶学方法测定甘油三酯(TG)和甘油含量,了解细胞脂解情况;采用Western-Blot法检测脂肪细胞中围脂滴蛋白A(PLIN1A)、脂肪甘油三酯脂肪酶(ATGL)、激素敏感性脂肪酶(HSL)和其磷酸化蛋白(p-HSL)的表达水平;双抗体夹心ABC-ELISA法检测细胞中环磷酸腺苷(cAMP)和蛋白激酶A(PKA)的浓度。结果 与空白组比较,ISO+sh-PLIN1组和ISO组脂滴变小,TG含量降低,甘油含量升高,有统计学意义(P<0.01)。同时,ISO+sh-PLIN1组和 sh-PLIN1组中ATGL 表达量均升高,有统计学意义(P<0.05)。ISO+sh-PLIN1转染组和ISO组中HSL、p-HSL、cAMP及 PKA 的表达量均上调(P<0.05)。结论 ISO促进脂解作用可能是cAMP/PKA通路介导,增加HSL和p-HSL的表达量来实现,而cAMP/PKA信号通路不能介导沉默PLIN1的促脂解作用。     

Comparison of vascular relaxation, lipolysis and glucose uptake by peroxisome proliferator-activated receptor-gamma activation in +db/+m and +db/+db mice

In this study, we determined the in vitro effect of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) activation on the aortic relaxation, lipolysis and insulin-induced [(3)H]-glucose uptake of the abdominal (omental) adipocytes of the non-diabetic (+db/+m) and obese/diabetic (+db/+db) mice. The expression of PPAR-gamma (mRNA and protein) in aorta and adipose tissues was evaluated and compared. Cumulative application of ciglitazone, pioglitazone and troglitazone (PPAR-gamma agonists) caused a concentration-dependent aortic relaxation (sensitive to 2-chloro-5-nitro-N-phenylbenzamide (GW9662) (1 microM, a selective PPAR-gamma antagonist) and N(omega)-nitro-l-arginine methyl ester (l-NAME) (20 microM, a nitric oxide synthase inhibitor)) with a maximum relaxation of approximately 30% (3 microM) in +db/+m mice, whereas no relaxation was observed in +db/+db mice. All PPAR-gamma agonists examined did not alter the basal lipolysis of both species, but forskolin caused a concentration-dependent lipolysis, with a greater magnitude observed in +db/+m mice. Insulin (0.1 and 1 microM) caused an enhancement of [(3)H]-glucose uptake into adipocytes with a greater magnitude in +db/+m mice. In contrast, none of the PPAR-gamma agonists tested (0.1, 1 and 10 microM) altered the basal and the insulin (0.1 microM)-induced [(3)H]-glucose uptake into adipocytes of both species. In addition, there was no difference in PPAR-gamma expression (mRNA and protein) in the aorta and adipose tissues between the species. In conclusion, our results demonstrate that PPAR-gamma is present in the abdominal (omental) adipose tissue and thoracic aorta. An acute activation of PPAR-gamma produced a small ( approximately 30%) aortic relaxation (nitric oxide/endothelium-dependent) of +db/+m mice. However, all PPAR-gamma agonists examined have no acute effect on lipolysis and the insulin-induced glucose uptake into adipocytes of both +db/+m and +db/+db mice.     

The adrenergic control of hepatic glucose and FFA metabolism in rainbow trout (Oncorhynchus mykiss): increased sensitivity to adrenergic stimulation with fasting

The adrenergic control of glucose and FFA release was studied in hepatocytes of rainbow trout (Oncorhynchus mykiss), which were either normally fed or fasted for 3 weeks. Isolated hepatocytes were incubated with adrenaline, noradrenaline, or isoprenaline (nonselective beta-agonist). Identification of the hepatic beta-adrenoceptor was combined with quantification of the difference in its affinity for adrenaline and noradrenaline. To identify the beta-adrenoceptor subtype, isoprenaline incubations were combined with atenolol (selective beta(1)-antagonist) or ICI 118,551 (selective beta2-antagonist). Stimulation of the beta-adrenoceptor resulted in mobilisation of glucose, which was inhibited by ICI 118,551 thus pointing to a beta2-subtype. The affinity of the beta2-adrenoceptor for isoprenaline and adrenaline (beta2-values of 8.3 and 7.9) was clearly higher than for noradrenaline (beta2-value of 6.5). This indicates that at physiological concentrations beta2-adrenoceptors in trout are mainly stimulated by adrenaline and not by noradrenaline. A significant effect of beta-adrenoceptor stimulation on the FFA release was also found, although only at high concentrations (i.e., 10(-6) and 10(-5)M). Again the beta2-adrenoceptor appeared to mediate the stimulation of hepatic FFA release. Upon fasting, both the basal glucose and FFA release were strongly decreased. The ratio between glucose and FFA release decreased from 15.4 to 4.3 upon fasting and at this ratio the energy output for both metabolites became equal. The mobilisation of FFA upon adrenergic stimulation was relatively conserved, namely -35% upon fasting, as opposed to -89% in mobilisation of glucose. This indicates that upon fasting FFA gain importance in hepatic metabolism. The hepatic sensitivity to adrenergic stimulation is enhanced upon fasting, as indicated by an increased beta2-value from 8.3 to 8.9.     

On the mechanism of some metabolic actions of dopamine

Summary The elevation of plasma glucose, glycerol and free fatty acids following the injection of dopamine in rats was reduced after treatment with desipramine or cocaine, while similar actions of noradrenaline on these metabolic parameters were enhanced after treatment with these drugs. Since the action of dopamine was also reduced by pretreatment with reserpine, we conclude that a significant part of the metabolic effects of dopamine is mediated by an indirect mechanism.Fellows of the Alexander von Humboldt-Foundation, W-Germany.     

Actions of dopamine in canine subcutaneous adipose tissue

Summary Subcutaneous adipose tissue in the inguinal region was isolated from surrounding tissues in fed female dogs and perfused with the dog's own blood.When dopamine (DA) was administered as a single i.a. injection in doses of 5 g, or higher, there was an increased vascular resistance and lipolysis. The vasoconstriction was abolished by phentolamine and phenoxybenzamine but was unaffected or potentiated by desipramine (5 mg i.a.). Desipramine abolished the lipolytic effect of DA as well as its capacity to increase the ³H-outflow from tissues prelabelled with L-³-H-noradrenaline.Infusion of DA (0.02 or 0.2 g/ml blood) had no effect on resting vascular resistance or on the glucose uptake. The higher DA concentration caused a significant increase in resting glycerol and free fatty acid release, while the lower was ineffective. DA did not significantly change the vasoconstrictor effect of sympathetic nerve stimulation (4 cps for 10 min) in either dose, but caused an increased lipolytic response.The results indicate that DA has many different actions in canine subcutaneous adipose tissue. Some of the possible mechanisms of action are discussed.     

Role of calcium and cyclic adenosine 3′,5′-monophosphate in the antilipolytic effect of tolbutamide and glibenclamide

Summary Tolbutamide (1 mM) and glibenclamide (0.1 mM) reduced lipolysis due to adrenaline, noradrenaline, isoprenaline, glucagon, and ACTH, whereas theophylline-induced glycerol production was only marginally affected. Furthermore, the effect of dibutyryl cyclic AMP on lipolysis was markedly decreased by low concentrations of both drugs. The antilipolytic action of glibenclamide and, to a smaller degree, of tolbutamide was highly dependent on the calcium concentration in the incubation medium used after preparation of fat cells in the absence of calcium. In the presence of isoprenaline as a lipolytic agent the antilipolytic effect of glibenclamide was progressively enhanced by raising the calcium concentration up to 1.2 mM. Howver, further elevation of calcium to 2.4 mM resulted in a loss of antilipolytic effect. Basal cyclic AMP levels of isolated fat cells were increased twofold by glibenclamide or tolbutamide. In contrast to the effect of glibenclamide, cyclic AMP levels in the presence of isoprenaline were further increased by tolbutamide. Low K _m cyclic AMP phosphodiesterase of fat cells was inhibited by tolbutamide as well as by glibenclamide. The results indicate that the antilipolytic action of sulfonylurea drugs is closely related to changes in calcium metabolism but independent of cyclic AMP levels.     

Dynamic strength training improves insulin sensitivity and functional balance between adrenergic alpha 2A and beta pathways in subcutaneous adipose tissue of obese subjects

Aims/hypothesis The aim of this study was to investigate whether dynamic strength training modifies the control of lipolysis, with particular attention paid to the involvement of the antilipolytic adrenergic alpha 2A receptor (ADRA2A) pathway. Methods Twelve obese men (age: 47.4±2.8 years; BMI: 32.7±0.9) were investigated during a 210-min euglycaemic–hyperinsulinaemic clamp conducted before and after 3 months of dynamic strength training. Before and during the third hour of the clamp, the lipolytic effect of a perfusion of isoproterenol or adrenaline (epinephrine) alone or associated with the ADRA2A antagonist phentolamine was evaluated using the microdialysis method of measuring extracellular glycerol concentration (EGC) in subcutaneous abdominal adipose tissue (SCAAT). In addition, biopsies of SCAAT were carried out before and after training to determine mRNA levels Results The training increased insulin sensitivity in adipose tissue. The decrease of EGC was more pronounced during the clamp conducted after the training period than during the clamp done in pre-training conditions. Before and after the training, catecholamines induced an increase in EGC, the increase being lower during the clamp on each occasion. The isoproterenol-induced increase in EGC was higher after the training. Adrenaline-induced lipolysis was potentiated by phentolamine after but not before the training. There were no training-induced changes in mRNA levels of key genes of the lipolytic pathway in SCAAT. Conclusions/interpretation In obese subjects, dynamic strength training improves whole-body and adipose tissue insulin responsiveness. It increases responsiveness to the adrenergic beta receptor stimulation of lipolysis and to the antilipolytic action of catecholamines mediated by ADRA2As.