Korean Circ J.  2024 Jan;54(1):43-56. 10.4070/kcj.2023.0120.

Entelon150^® (Vitis vinifera Seed Extract) Attenuates Degenerative Changes in Intravascular Valve Prostheses in Rabbits

• Lee JS
• Seo J
• Kim S
• Rahman M
• Shin HJ

• KMID: 2550608
• DOI: http://doi.org/10.4070/kcj.2023.0120

Abstract

Background and Objectives
The therapeutic strategy for inflammation and degenerative calcification is of utmost importance for bioprosthetic heart valve (BHV) implanted patients. The purpose of this study was to compare the anti-inflammatory and anti-calcification effects of Entelon150^® (grape seed extract), losartan, and rosuvastatin, in a rabbit model of intravascular BHV leaflet implantation in bovine pericardium.
Methods
A total of 28 rabbits were implanted with BHV leaflet in the external jugular veins. The Entelon150^® group was administered 7.7 mg/kg Entelon150^® twice daily for 6 weeks after surgery. The losartan and rosuvastatin groups received 5.14 mg/kg and 1 mg/kg, respectively, once per day. The control group received 1 ml of saline once daily. And then, calcium concentration was measured in the implanted BHV, and histological and molecular analyses were performed on the surrounding tissues.
Results
The calcium content of the implanted tissue in the Entelon150^® group (0.013±0.004 mg/g) was lower than that in the control group (0.066±0.039 mg/g) (p=0.008). The losartan (0.024±0.016 mg/g, p=0.032) and rosuvastatin (0.022±0.011 mg/g, p=0.032) groups had lower calcium content than the control group, and higher tendency than the Entelon150^® group. Immunohistochemistry revealed that the expressions of bone morphogenic protein 2 (BMP2), S-100, and angiotensin II type 1 receptor in the Entelon150^® group showed lower tendency than those in the control group. The protein expression levels of BMP2 were reduced in the Entelon150^® group compared with those in the control group.
Conclusions
Entelon150^® exhibited a significant effect, similar to other drugs, in reducing calcification and inflammation in the intravascular bovine pericardium.

Keyword

Vitis vinifera; Heart valve prosthesis, Inflammation; Vascular calcification, Losartan; Rosuvastatin

Figure

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  Figure 1 Surgical images of intravascular BHV implantation. (A) Bovine pericardium BHV fixed to the inner wall of the vein. (B) Vascular incision site being sutured and confirmed blood flow.BHV = bioprosthetic heart valve.

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  Figure 2 Schematic diagram of the anatomy of New Zealand white rabbit jugular vein.

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  Figure 3 Calcium concentration in the implanted BHV tissue in each group. Calcium levels were measured 6 weeks after implantation. The calcium levels in the control group were significantly higher than those in the Entelon, losartan, and rosuvastatin groups. Data are presented as mean±standard deviation.BHV = bioprosthetic heart valve.

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  Figure 4 Effects of Entelon150, losartan, and rosuvastatin on inflammatory cell infiltrationCompared to the control group, the Entelon-, losartan-, and rosuvastatin-treated groups displayed less infiltration of inflammatory cells (giant cells, macrophages, heterophils, and lymphocytes) into the tissue surrounding the implanted BHV and within the fibers of the implanted BHV; however, no statistically significant difference was found. An intravenous bovine pericardial implant (asterisk) was detected in the microscopic images. 40× scale bar indicates 50 µm. 200× scale bar indicates 100 µm.BHV = bioprosthetic heart valve.

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  Figure 5 Effects of each drugs on the expression of calcific, inflammatory, and myofibroblastic markers. Data are presented as mean±standard deviation. 400× scale bar indicates 50 µm.α-SMA = alpha smooth muscle actin; AIIR = angiotensin II type 1 receptor; BMP2 = bone morphogenic protein 2.

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  Figure 6 Effects of each drug on the calcific and inflammatory protein expression. Data are reported as mean±standard deviation.BMP2 = bone morphogenic protein 2; IL-6 = interleukin-6; OPN = osteopontin.

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