To establish an HPLC method for determining components in monocrotalinum liposomes. The results showed a good linear relationship in monocrotalinum liposomes within the concentration range between 1.6-102.4 mg x L(-1) (r = 0.999 8), with RSDs of intra-day precision, inter-day precision, stability and reproducibility of 0.61%, 0.92%, 1.7%, 1.6%, respectively. The recovery rate of monocrotaline was (99.96 +/- 0.50)%. These data indicated that the HPLC method could accurately determine components in monocrotalinum liposomes. Meanwhile, the microcolumn centrifugation method was established to determine the entrapment efficiency of components in monocrotalinum liposomes. As a result, the recovery rate and the blank liposome recovery of free components were (94.44 +/- 0.77)%, (95.86 +/- 0.68 )%, respectively. According to the parallel determination of the entrapment efficiency of three monocrotaline liposomes, their RSD was 4.0%. The data indicated that the microcolumn centrifugation method was an accurate and feasible method for determining the entrapment efficiency of monocrotaline liposomes.
Chromatography, High Pressure Liquid ; methods ; Drug Carriers ; chemistry ; Drug Compounding ; Liposomes ; chemistry ; Monocrotaline ; chemistry

Animals ; Hypertension, Pulmonary ; chemically induced ; physiopathology ; Monocrotaline ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Thrombomodulin ; metabolism

OBJECTIVETo observe the pathological changes in the myocardial and pulmonary tissues in miniature pigs with chronic pulmonary hypertension induced by monocrotaline (MCT).

METHODSTwelve male miniature pigs (weigh 15.0-18.0 kg, aged 4.0-4.5 months) were examined for baseline mean pulmonary artery pressure (mPAP), followed by intraperitoneal injection of 10.0 mg/kg MCT in 10 randomly selected pigs. The mean pulmonary artery pressure at 4 and 8 weeks were determined, and the pathological changes in the myocardial and pulmonary tissues were observed.

RESULTSThe baseline mPAP of normal miniature pigs was 15.19∓0.70 mmHg. At 4 and 8 weeks after MCT injection, the sPAP and dPAP were 19.69∓2.47 mmHg and 25.62∓4.88 mmHg, respectively, and the mPAP increased significantly compared with that of the normal control group (P<0.01). Obvious pathological changes such as pulmonary hypertension and right ventricular hypertrophy were found in the pigs 4 weeks after MCT injection, and at 8 weeks, significant pathological changes occurred including right ventricular fibrosis and thickening of the tunica media of the pulmonary artery.

CONCLUSIONMCT can cause pulmonary hypertension in miniature pigs 8 weeks after drug administration, shown as increased pulmonary artery pressure and pulmonary vascular remodeling.

Animals ; Hypertension, Pulmonary ; chemically induced ; pathology ; Lung ; pathology ; Male ; Monocrotaline ; adverse effects ; Myocardium ; pathology ; Swine ; Swine, Miniature

OBJECTIVETo investigate the fetotoxicity of monocrotaline.

METHODMouse whole embryo culture (WEC) was applied. Post-implantation (8.5 d) mouse embryos were isolated from their mothers and put into the medium of immediately centrifuged serum (ICS) prepared from rats. Different concentrations of monocrotaline (100, 50, 25, 12.5 mg x L(-1)) were added into the WEC. Development (yolk sac diameter, crown-rump length, head length, somite number) and organic morphodifferentiation (yolk sac circulation, allantois, embryonic flexion, heart, brain, optic-otic-olfactory organ, branchial arch, maxillary, mandible, bud) of embryos were observed at 48 h after treatment.

RESULTObvious fetotoxicity could be observed in various monocrotaline treatment groups in a dose-dependent manner. Development of embryos was delayed significantly at dose 12.5-100 mg x L(-1). Malformations were shown in all organic morphodifferentiation indice, especially in opti-otic organ, mandible and bud.

CONCLUSIONMonocrotaline had obvious fetotoxicity in vitro WEC, indicating that exposure of pregnant mice to monocrotaline may have potential risk on fetus.

Animals ; Cell Differentiation ; drug effects ; Culture Media ; Embryo, Mammalian ; drug effects ; physiology ; Female ; Male ; Mice ; Monocrotaline ; toxicity

OBJECTIVE: To study the protective effects of ginkgo biloba extract on the right ventricular hypertrophy. METHODS: Seventy-two SD male rats were randomly divided into 3 groups: control group(CON), monocrotaline-induced right ventricular hypertrophy group (MCT) and ginkgo biloba extract treated group (EGB) (n=24 in each group). Group MCT and group EGB were intraperitoneally injected with 2%MCT at the dose of 60 mg /kg on the first day. From the second day, group MCT was injected with 2 ml 0.9% sodium chloride, and 60 mg/kg ginkgo leaf extract was administered to the stomach in group EGB. The control group was injected with 2 ml 0.9% sodium chloride on the first day. After 3 weeks, in each group,cardiac hemodynamic changes were measured, heart weight index was calculated, and myocardial pathological changes were observed by HE staining. The expression of TRPC6 was detected by real-time polymerase chain reaction (real-time -PCR) and Western blot. RESULTS: Compared with the control group, the right ventricular systolic pressure (RVSP) was increased significantly in the MCT group(P＜0.01), the maximum or decline rate of descent (RV ±dp/dt) of the right ventricle pressure was increased significantly(P＜0.01), while the EGB group had the same trend as all the indexes in the group MCT, but the amplitude of all indicators in group EGB were decreased significantly than those of group MCT(P＜0.01), and the right ventricular hypertrophy index (RVMI) in group EGB was significantly lower than that in group MCT(P＜0.01).Group MCT showed typical myocardial hypertrophy performance by HE staining, and the right ventricular myocytes in group EGB were significantly improved than that in group MCT, and the mRNA and protein expression levels of TRPC6 in the right ventricle of group MCT and group EGB were increased(P＜0.01), while the EGB group was significantly lower than that of the MCT group(P＜0.01). CONCLUSION Ginkgo biloba extract may inhibit the signal pathway of CaN / NFAT in cardiac myocytes by reducing the expression of TRPC6 protein, and then play an early protective effect on myocardial hypertrophy.
Animals ; Hypertrophy, Right Ventricular ; chemically induced ; drug therapy ; Male ; Monocrotaline ; Plant Extracts ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

The pathophysiological implications of aldosterone and adrenomedullin in the cardiac ventricular hypertrophy were examined. Male Sprague-Dawley rats were treated with deoxycorticosterone acetate (DOCA)-salt and monocrotaline (MCT) to selectively elicit left and right ventricular (LV, RV) hypertrophy, respectively. The mRNA expression of aldosterone synthase and adrenomedullin in LV and RV was determined by reverse transcription-polymerase chain reaction. The expression of aldosterone synthase and adrenomedullin was increased in LV, while not altered significantly in RV of DOCA-salt-treated rats. On the contrary, the expression was not significantly altered in LV, but increased in RV of MCT-treated rats. The enhanced expression of aldosterone synthase may be causally related with the development of ventricular hypertrophy, and the increased expression of adrenomedullin may act as a counter-regulatory mechanism.
Adrenomedullin* ; Aldosterone Synthase* ; Aldosterone* ; Animals ; Desoxycorticosterone ; Humans ; Hypertrophy ; Hypertrophy, Right Ventricular* ; Male ; Monocrotaline ; Rats* ; Rats, Sprague-Dawley ; RNA, Messenger

The present study was aimed to explore pathophysiological implications of nitric oxide in the development of left and right ventricular hypertrophy. To induce selective left and right ventricular hypertrophy, rats were made two-kidney, one clip (2K1C) hypertensive and treated with monocrotaline (MCT), respectively. Six weeks later, the hearts were taken and their ventricular tissue mRNA and protein expression of endothelial constitutive isoform of nitric oxide synthase (NOS) were determined by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. In 2K1C hypertensive rats, the expression of NOS mRNA was increased in parallel with its proteins in the left ventricle, but not in the right ventricle. In MCT-treated rats, the expression of NOS mRNA and proteins were proportionally increased in the right ventricle, but not in the left ventricle. These results suggest that the expression of NOS is specifically increased in association with the ventricular hypertrophy, which may be a mechanism counteracting the hypertrophy.
Animals ; Blotting, Western ; Cardiomegaly ; Heart ; Heart Ventricles ; Hypertrophy ; Hypertrophy, Right Ventricular* ; Monocrotaline ; Nitric Oxide Synthase ; Nitric Oxide* ; Rats ; RNA, Messenger

PURPOSE: To examine the effect of bosentan, a dual endothelin receptor (ER) antagonist, on the development of monocrotaline (MCT)-induced pulmonary hypertension in rats by especially focusing on the pulmonary vascular morphology changes. METHODS: Sprague-Dawley rats were treated as follows: controls received a subcutaneous saline injection, MCT-treated rats received a subcutaneous MCT injection, and bosentan-treated rats received a MCT injection followed by treatment with bosentan (20 mg/kg/day). To assess the effects of ER blockade on the time course, the animals were exsanguinated, and their hearts and lungs were dissected after 7, 14, or 28 days. RESULTS: The mean body weights of the MCT- and bosentan-treated rats were significantly lower than that of the control rats on days 7, 14, and 28. Bosentan administration significantly inhibited the progression of right ventricular hypertrophy on day 28 (right ventricle/[left ventricle+septum]: 0.71+/-0.10 in MCT-treated rats vs. 0.49+/-0.09 in bosentan-treated rats; P<0.05). Quantitative analysis of peripheral pulmonary arteries revealed that the increase in medial wall thickness after MCT injection was significantly attenuated in the bosentan-treated rats on day 28 (49.96+/-10.06% in MCT-treated rats vs. 47.09+/-10.48% in bosentan-treated rats; P<0.05). In addition, the increase in the number of intra-acinar muscular arteries after MCT injection was reduced by bosentan on days 14 and 28. CONCLUSION: Bosentan administration in intermediate doses exerts inhibitory effects on lung vascular hypertrophy and right ventricular hypertrophy during the development of MCT-induced pulmonary hypertension in rats.
Animals ; Arteries ; Body Weight ; Endothelins ; Heart ; Hypertension, Pulmonary ; Hypertrophy ; Hypertrophy, Right Ventricular ; Lung ; Monocrotaline ; Pulmonary Artery ; Rats ; Rats, Sprague-Dawley ; Receptors, Endothelin ; Sulfonamides

BACKGROUND AND OBJECTIVES: Endothelin (ET)-1, a potent endothelium-derived vasoconstrictor peptide, has a potential pathophysiologic role in pulmonary hypertension. Bosentan, a dual ET receptor (ET(A)/ET(B)) antagonist, is efficacious in treatment of pulmonary hypertension. The objectives of this study were to investigate the expression of ET-1 and ET receptor A (ERA) genes and to evaluate the effect of bosentan in monocrotaline (MCT)-induced pulmonary hypertension. MATERIALS AND METHODS: Four-week-old male Sprague-Dawley rats were treated as follows: control (n=36), subcutaneous (sc) injection of saline; MCT (n=36), sc injection of MCT (60 mg/kg); and bosentan (n=36), sc injection of MCT (60 mg/kg) plus 25 mg/kg/day bosentan orally. RESULTS: Serum ET-1 concentrations in the MCT group were higher than the control group on day 28 and 42. Quantitative analysis of peripheral pulmonary arteries revealed that the increase in medial wall thickness after MCT injection was significantly attenuated in the bosentan group on day 28 and 42. In addition, the increase in the number of intra-acinar muscular arteries after MCT injection was reduced by bosentan on day 14, 28 and 42. The levels of ET-1 and ERA gene expression were significantly increased in the MCT group compared with control group on day 5, and bosentan decreased the expression of ET-1 on day 5. CONCLUSION: ET-1 contributes to the progression of cardiopulmonary pathology in rats with MCT-induced pulmonary hypertension. Administration of bosentan reduced ET-1 gene expression in MCT-induced pulmonary hypertension in rats.
Animals ; Arteries ; Endothelin-1 ; Endothelins ; Gene Expression ; Humans ; Hypertension, Pulmonary ; Male ; Monocrotaline ; Pulmonary Artery ; Rats ; Rats, Sprague-Dawley ; Receptors, Endothelin ; Sulfonamides

PURPOSE: Pulmonary arterial hypertension (PAH) is a fatal disease which is characterized by an increase in pulmonary arterial pressure leading to increases in right ventricular afterload. Human umbilical cord blood derived-mesenchymal stem cells (hUCB-MSCs) administered via the jugular vein have been previously shown to improve PAH by reversal treatment. However, the effect of low dosage and transfusion timing of hUCB-MSCs on PAH has not yet been clearly established. Obviously, low dosage treatment can lead to a reduction in costs. This is the first study on early transfusion effect. MATERIALS AND METHODS: This study was divided into two parts. The first part is an investigation of dose-dependent effect. hUCB-MSCs were administered into 3 groups of rats (UA: 3×10⁶ cells, UB: 1.5×10⁶ cells, UC: 3×10⁵ cells) via the external jugular vein at week 1 after monocrotaline (MCT) injection. The second part is a search for optimal treatment timing in 3×10⁵ cells dose of hUCB-MSCs administered at day 1 for UD group (low dose of hUCB-MSCs at day 1), at day 1 and week 1 for the UE group (dual transfusion of low dose of hUCB-MSCs at day 1 and week 1) and at 1 week for the UF group (reversal treatment of low dose hUCB-MSC at week 1) after MCT injection. RESULTS: The administration of 3×10⁵ hUCB-MSCs was as effective as the 3×10⁶ dose in decreasing mean right ventricle (RV) pressure and pulmonary pathological changes. Early treatment with hUCB-MSCs improved mean RV pressure, pulmonary pathological changes and heart collagen 3 protein expression levels in PAH. CONCLUSION: Low-dose early treatment of hUCB-MSCs is as effective as a high dose treatment of hUCB-MSCs in improving PAH although dual or reversal treatment is still more effective.
Animals ; Arterial Pressure ; Collagen ; Fetal Blood ; Heart ; Heart Ventricles ; Humans ; Hypertension ; Hypertension, Pulmonary ; Jugular Veins ; Mesenchymal Stromal Cells ; Monocrotaline ; Rats* ; Stem Cells*