Stable Gastric Pentadecapeptide BPC 157 May Counteract Myocardial Infarction Induced by Isoprenaline in Rats

Abstract

We revealed that the stable gastric pentadecapeptide BPC 157, a useful peptide therapy against isoprenaline myocardial infarction, as well as against isoprenaline myocardial reinfarction, may follow the counteraction of the recently described occlusion-like syndrome, induced peripherally and centrally, which was described for the first time in isoprenaline-treated rats. BPC 157 (10 ng/kg, 10 µg/kg i.p.), L-NAME (5 mg/kg i.p.), and L-arginine (200 mg/kg i.p.) were given alone or together at (i) 30 min before or, alternatively, (ii) at 5 min after isoprenaline (75 or 150 mg/kg s.c.). At 30 min after isoprenaline 75 mg/kg s.c., we noted an early multiorgan failure (brain, heart, lung, liver, kidney and gastrointestinal lesions), thrombosis, intracranial (superior sagittal sinus) hypertension, portal and caval hypertension, and aortal hypotension, in its full presentation (or attenuated by BPC 157 therapy (given at 5 min after isoprenaline) via activation of the azygos vein). Further, we studied isoprenaline (75 or 150 mg/kg s.c.) myocardial infarction (1 challenge) and reinfarction (isoprenaline at 0 h and 24 h, 2 challenges) in rats (assessed at the end of the subsequent 24 h period). BPC 157 reduced levels of all necrosis markers, CK, CK-MB, LDH, and cTnT, and attenuated gross (no visible infarcted area) and histological damage, ECG (no ST-T ischemic changes), and echocardiography (preservation of systolic left ventricular function) damage induced by isoprenaline. Its effect was associated with a significant decrease in oxidative stress parameters and likely maintained NO system function, providing that BPC 157 interacted with eNOS and COX2 gene expression in a particular way and counteracted the noxious effect of the NOS-blocker, L-NAME.

Keywords: L-NAME; L-arginine; isoprenaline; myocardial infarction; occlusion-like syndrome; oxidative stress; pentadecapeptide BPC 157; rats.

Figure 1

Presentation of the early occlusion-like syndrome in isoprenaline (75 mg/kg sc) treated rats at 30 min after isoprenaline administration. Medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline 5.0 mL/kg (C) i.p.) was given 5 min after isoprenaline. Heart, lung, liver and kidney lesions microscopic scoring (a); gastrointestinal lesion microscopic scoring (b); brain lesion microscopic scoring (c); blood (superior sagittal sinus (SSS), portal vein (PV), inferior caval vein (ICV), abdominal aorta (AA)) pressure (d); thrombus mass (portal vein (PV), inferior caval vein (ICV), superior sagittal sinus (SSS), abdominal aorta (AA) and superior mesenteric artery (SMA)) (e); and vessels (inferior caval vein (ICV), superior mesenteric vein (SMV), azygos vein (AV) and brain relative volume (f) assessments were carried out. Ten rats per each experimental group. Means ± SD, min/med/max, * p ˂ 0.05, at least, vs. control.

Figure 2

Regular timeline (30 min after isoprenaline) of vessel presentation in the isoprenaline-treated rats, disturbed (congested or collapsed) in controls (a–c) and normalized in BPC 157-treated rats (d–f), including the superior mesenteric vein (white arrows), inferior caval vein (yellow arrows), azygos vein (blue arrows) and thoracic and aorta (red arrows). Upon BPC 157 therapy, recruitment of the defensive pathways counteracted congestion of the superior mesenteric vein and inferior caval vein and counteracted failure of the azygos vein and thoracic and abdominal aorta timeline (d–f).

Figure 3

Regular timeline (30 min after isoprenaline) of organ lesion presentations in the isoprenaline-treated rats, in controls (a–d) and lesion counteraction in BPC 157-treated rats (e–h). Heart dilatation (a), brain swelling (b,c) and stomach lesions (d) in control-isoprenaline rats. Upon BPC 157 therapy, these lesions were markedly attenuated or eliminated (e–h).

Figure 4

Heart and lung tissue damage. Isoprenaline 75 mg/kg at 30 min after challenge. Control rats (a,b). Myocardial congestion and confluent areas of subendocardial ischemic myocytes in the control rats (circle). Control rats (c). Marked congestion of the lung parenchyma in the control rats. BPC 157-treated rats (d,e). Mild myocardial congestion and only a small area of subendocardial ischemic myocytes (circle) in the BPC 157-treated rats (f). No changes in lung tissue in the BPC 157-treated rats. (HE staining; (a,d) magnification 400×; scale bar 100 μm; (b,c,e,f) magnification 100×; scale bar 200 μm).

Figure 5

Illustrative features of liver and kidney injury. Isoprenaline 75 mg/kg at 30 min after challenge. Control rats (a,b). (a) Marked dilatation and congestion of blood vessels in the portal tracts as well as dilatation of central veins and sinusoids. (b) Conspicuous dilatation of blood vessels and congestion in the kidney tissue, as well as glomeruli. BPC 157-treated rats (c,d). (c) No changes in liver tissue. (d) Mild dilatation and congestion of blood vessels in the kidney tissue. (HE staining; magnification 100×; scale bar 200 μm).

Figure 6

Gastrointestinal tract injury. Isoprenaline 75 mg/kg at 30 min after challenge. Control rats ((a) (stomach), (b) (small intestine), (c) (large intestine)). Congestion in the upper and lower digestive tract with a tendency pronounced in the lower digestive tract. BPC 157-treated rats ((d) (stomach), (e) (small intestine), (f) (large intestine)). No changes in the stomach and small intestine with mild congestion of the colon. (HE staining; magnification 200×; scale bar 200 μm).

Figure 7

Neuropathological changes of cerebral area. Isoprenaline 75 mg/kg at 30 min after challenge. Control group (a,b). (a) Large areas of intracerebral hemorrhage. B. Intraventricular hemorrage in the 3rd ventricle. BPC 157 treated rats (c,d). (c) Minor intracerebral hemorrhage. (d) No hemorrhage in the 3rd ventricle. (HE; magnification 100×; scale bar 200 μm).

Figure 8

Neuropathological features of cerebral and celebellar cortex. Isoprenaline 75 mg/kg at 30 min after challenge. Control rats (a,b). (a) Some karyopyknotic cells in cerebral cortex (black arrows). (b) Moderate karyopyknosis and degeneration of Purkinje cells of the cerebellar cortex. BPC 157-treated rats (c,d). (c) No karyopyknotic cells in cerebral cortex. (d) Only a few karyopyknotic and degenerated Purkinje cells of the cerebellar cortex were found. (HE; magnification 400×; scale bar 50 μm).

Figure 9

Neuropathological features of the hippocampus and hypothalamus. Isoprenaline 75 mg/kg at 30 min after challenge. Control rats (a,b). (a) Marked karyopyknosis of pyramidal cells of the hippocampus (black arrows). (b) Rare karyopyknotic cells in hypothalamic area. BPC 157-treated rats (c,d). (c) Small number karyopyknosis of pyramidal cells of the hippocampus. (d) No changes in hipothalamic area. (HE staining; magnification 400×; scale bar 100 μm).

Figure 10

(A,B) Myocardial infarction (one isoprenaline challenge) and re-infarction (two isoprenaline subsequent challenges) in isoprenaline-treated rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)). Serum values of cardioselective enzymes A-CK-MB (UI/L) and B-CK (UI/L). Rats treated with the smaller dose of isoprenaline, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Rats treated with the higher dose of isoprenalines, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)) or L-NAME (5 mg/kg i.p. (N)), L-arginine (200 mg/kg i.p. (A)) (horizontal line bars) or these agents in combination (L-NAME + L-arginine (NA), L-NAME + BPC 157 (NB), L-arginine + BPC 157 (AB), L-NAME + L-arginine + BPC 157 (NAB)) (dashed horizontal lines bars). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT), prophylactic regimen (a–d), or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (e–h)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 11

(A,B) Myocardial infarction (one isoprenaline challenge) and re-infarction (two isoprenaline subsequent challenges) in isoprenaline- treated rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)). Serum values of cardioselective enzymes A-LDH (UI/L) and B-cTnT ng/mL. Rats treated with the smaller dose of isoprenaline, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Rats treated with the higher dose of isoprenaline, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)) or L-NAME (5 mg/kg i.p. (N)), L-arginine (200 mg/kg i.p. (A)) (horizontal line bars) or these agents combination (L-NAME + L-arginine (NA), L-NAME + BPC 157 (NB), L-arginine + BPC 157 (AB), L-NAME + L-arginine + BPC 157 (NAB)) (dashed horizontal lines bars). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (a–d)), or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (e–h)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 12

Myocardial infarction (one isoprenaline challenge) and re-infarction (two isoprenaline subsequent challenges) in isoprenaline-rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)), gross heart lesions presentation, A- INFARCTED HEART SURFACE. Isoprenaline-rats treated with the smaller dose, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Isoprenaline-rats treated with the higher dose, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)) or L-NAME (5 mg/kg i.p. (N)), L-arginine (200 mg/kg i.p. (A)) (horizontal line bars) or these agents in combination (L-NAME + L-arginine (NA), L-NAME + BPC 157 (NB), L-arginine + BPC 157 (AB), L-NAME + L-arginine + BPC 157 (NAB)) (dashed horizontal lines bars). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (a–d)), or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (e–h)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 13

Myocardial infarction (one isoprenaline challenge) and re-infarction (two isoprenaline subsequent challenges) in isoprenaline-treated rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)) and heart lesion microscopy presentation, A- HEART MICROSCOPY. Rats treated with the smaller dose of isoprenaline, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Rats treated with the higher dose of isoprenaline, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)) or L-NAME (5 mg/kg i.p. (N)), L-arginine (200 mg/kg i.p. (A)) (horizontal line bars) or these agents in combination (L-NAME + L-arginine (NA), L-NAME + BPC 157 (NB), L-arginine + BPC 157 (AB), L-NAME + L-arginine + BPC 157 (NAB)) (dashed horizontal lines bars). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (a–d)), or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (e–h)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 14

Illustrative gross presentation of anterior heart surface in rats at 24 h after isoprenaline. Rats challenged once with isoprenaline 150 mg/kg s.c., mediation at 30 min before isoprenaline. (a) Significant myocardial infarction in controls that received saline (5 mL/kg i.p.). (b) L-NAME (5 mg/kg i.p.)-pretreated rats presented with large infarcted area extended to the anterior, lateral, apical and inferior heart surfaces. (c) L-arginine (200 mg/kg i.p.)-pretreated rats presented with a lesser infracted area. (d) BPC 157 (10 μg/kg i.p.)-pretreated rats presented with no visible infracted area.

Figure 15

Microscopy presentation of heart injury. Rats challenged once with isoprenaline 150 mg/kg s.c., mediation at 30 min before isoprenaline (a–d). Upper 24 h. (a,c) Single and small groups of necrotic myocites without inflammation (irregular circles) (magnification 400×, scale bar 20 μm) (saline (5 mL/kg i.p, left)) (a); single and small groups of necrotic myocites without inflammation (magnification 400x, scale bar 20 μm) (BPC 157 (10 μg/kg i.p., right)) (c). Low (b,d). Pronounced inflammatory infiltrate with necrotic myocites (magnification 400×, scale bar 20 μm) (saline (5 mL/kg i.p, left)) (b). Moderate inflammatory infiltrate affecting up to half of the septal thickness (magnification 400×, scale bar 20 μm) (BPC 157 (10 μg/kg i.p., right)). (d). (HE staining).

Figure 16

(A,B) Myocardial infarction (one isoprenaline challenge) and re-infarction (two isoprenaline subsequent challenges) in isoprenaline-treated rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)). Gross heart lesions presentation; A–ST-ELEVATION/DEPRESSION (mV), B–HEART FREQUENCY (beats/min). Rats treated with the smaller dose of isoprenaline, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Rats treated with the higher dose of isoprenaline, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)) or L-NAME (5 mg/kg i.p. (N)), L-arginine (200 mg/kg i.p. (A)) (horizontal line bars) or these agents in combination (L-NAME + L-arginine (NA), L-NAME + BPC 157 (NB), L-arginine + BPC 157 (AB), L-NAME + L-arginine + BPC 157 (NAB)) (dashed horizontal lines bars). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (a–d)), or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (e–h)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 17

ECG presentation. Rats challenged once with isoprenaline 150 mg/kg s.c., medication at 30 min before isoprenaline. (a) ST elevation in all 3 standard leads in controls that received saline (5 mL/kg ip) 24 h after first isoprenaline challenge. (b) L-NAME (5 mg/kg i.p.) medication at 30 min before isoprenaline presented with ST elevation in all 3 standard leads. (c) L-arginine (200 mg/kg i.p.) presented with less expressed ischemic ECG changes at 24 h after initial isoprenaline dose. (d) BPC 157 (10 µg/kg i.p.) presented with no ST-T ischemic changes.

Figure 18

(A,B) Myocardial infarction (1 isoprenaline challenge) and re-infarction (2 isoprenaline subsequent challenges) in isoprenaline-rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)). Gross heart lesions presentation. A–ECHOCARDIOGRAPHY, PRETREATMENT; B–ECHOCARDIOGRAPHY, POST-TREATMENT. Isoprenaline-rats treated with the smaller dose, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Isoprenaline-rats treated with the higher dose, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (EF LV (%), 24 h, infarction (a), 48 h reinfarction (b), LDV (mL) diastolic volume, 24 h, infarction (c), 48 h reinfarction (d), LVSV (mL), end-diastolic volume, 24 h, infarction (e), 48 h reinfarction (f)) or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (EF LV (%), 24 h, infarction (a), 48 h reinfarction (b), LDV (mL) diastolic volume, 24 h, infarction (c), 48 h reinfarction (d), LVSV (mL), end-diastolic volume, 24 h, infarction (e), 48 h reinfarction (f)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 18

(A,B) Myocardial infarction (1 isoprenaline challenge) and re-infarction (2 isoprenaline subsequent challenges) in isoprenaline-rats (75 mg/kg sc (gray bars), 150 mg/kg sc (white bars)). Gross heart lesions presentation. A–ECHOCARDIOGRAPHY, PRETREATMENT; B–ECHOCARDIOGRAPHY, POST-TREATMENT. Isoprenaline-rats treated with the smaller dose, 75 mg/kg sc (gray bars, italic letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Isoprenaline-rats treated with the higher dose, 150 mg/kg sc (white bars, normal letters), received medication (BPC 157 (10 ng/kg (b), 10 µg/kg (B) i.p. or saline (5 mL/kg i.p. (C)). Therapy was given (i) 30 min before isoprenaline (PRETREATMENT, prophylactic regimen (EF LV (%), 24 h, infarction (a), 48 h reinfarction (b), LDV (mL) diastolic volume, 24 h, infarction (c), 48 h reinfarction (d), LVSV (mL), end-diastolic volume, 24 h, infarction (e), 48 h reinfarction (f)) or, alternatively, (ii) at 5 min after isoprenaline (75 mg/kg s.c. or 150 mg/kg s.c.), at day 1 and at day 2 (POST-TREATMENT, therapeutic regimen (EF LV (%), 24 h, infarction (a), 48 h reinfarction (b), LDV (mL) diastolic volume, 24 h, infarction (c), 48 h reinfarction (d), LVSV (mL), end-diastolic volume, 24 h, infarction (e), 48 h reinfarction (f)). Ten rats per each experimental group. * p < 0.05 vs. control, at least.

Figure 19

MDA levels in heart tissue. MDA levels in the heart tissue of isoprenaline-treated rats at 24 h after challenge (isoprenaline 75 mg/kg sc) (gray bars), determined by quantifying thiobarbituric acid (TBA) reactivity as malondialdehyde (MDA) equivalents. BPC 157 (10 µg/kg ip) (B) or an equal volume of saline (5 mL/kg ip) (controls) (C) was applied to the isoprenaline-treated rats before or after the challenge. Ten rats per each experimental group. * p < 0.05 vs. saline; # p < 0.05 healthy (dashed bar) vs. isoprenaline-treated rats (H).

Figure 20

Summary of the background of the isoprenaline effects (red frameworks) and therapy potential of BPC 157 application (blue frameworks) [14,15,16,17,18,19,20,21,22,23]. It is likely that isoprenaline, via subcutaneous administration, indicates irremovable (endothelial) lesions (red frameworks), in which the full peripheral and central syndromes appear in addition to each other (red frameworks) as also noted with intragastric absolute alcohol-induced, over-dose lithium-induced and maintained intra-abdominal hypertension-induced occlusion-like syndrome (red dashes frameworks) [21,22,23]. Due to the prime peripheral or central lesion, these lesions should appear as an essential cause–consequence cycle (red dashes frameworks) [14,15,16,17,18,19,20,21,22,23]. BPC 157 therapy means the activation of the collateral pathways when confronted with vessel occlusion or other vessel failure, and a consequent severe or even deadly syndrome, which is a shared syndrome, peripherally and centrally, and has been recently revealed, which may bring the theory into additional practical application (blue framework). Using the BPC 157 regimen, we commonly observed the counteraction of intracranial (superior sagittal sinus) hypertension, portal and caval hypertension, aortal hypotension, and progressing venous and arterial thrombosis peripherally and centrally; furthermore, ECG disturbances were attenuated. Multiple organ lesions, were also attenuated, including those of the heart, lung, liver, kidney, and gastrointestinal tract, in particular, as well as brain lesions. Oxidative stress in tissues were also attenuated (blue dashes framework). Thus, this may be suitable background for the counteraction of the early isoprenaline-induced occlusion-like syndrome, and thereby, isoprenaline-induced myocardial infarction and reinfarction.