Anat Cell Biol.  2022 Jun;55(2):205-216. 10.5115/acb.21.187.

Antioxidant, anti-inflammatory, and anti-fibrotic properties of olive leaf extract protect against L-arginine induced chronic pancreatitis in the adult male albino rat

• Romeh GH
• El-Safty FENAH
• El-Mehi AES
• Faried MA

• KMID: 2531207
• DOI: http://doi.org/10.5115/acb.21.187

Abstract

Chronic pancreatitis (CP) is an inflammatory disease affects the pancreas with upcoming fibrosis and notable parenchymal destruction. CP poses a high risk for pancreatic carcinoma. The present study aimed to investigate, for the first time up to our knowledge, the effect of olive leaf extract on L-arginine induced CP with referral to some of its underlying mechanisms. Forty adult male albino rats were divided equally into four groups; control, olive leaf extract treated (200 mg/ kg orally once daily), CP group (300 mg L-arginine/100 g body weight intraperitoneally, once daily for 3 weeks then every 3 days for the subsequent 3 weeks), and CP treated with olive leaf extract group. At the end of the experiment, body weight, serum glucose, serum insulin, homeostatic model assessment of insulin resistance (HOMA-IR), serum amylase and lipase as well as tissue superoxide dismutase (SOD), and malondialdehyde (MDA) levels were assessed. Pancreatic tissues were subjected to histological and immuno-histochemical studies. The CP group revealed significant decrease in body weight and increase in serum glucose, serum insulin, HOMA-IR score, serum amylase, and serum lipase levels. Significant increase in MDA level and decrease in SOD level were detected. Marked degenerative changes and fibrosis were detected. Upregulation of alpha smooth muscle actin (α-SMA), transforming growth factor beta (TGF-β), caspase-3, and interleukin-6 (IL-6) immunoreactions were implicated in CP pathogenesis. Olive leaf extract alleviated all the examined parameters via itsantioxidant, anti-inflammatory, and anti-fibrotic properties. Olive leaf extract can protect against CP and restore pancreatic functions.

Keyword

Olive leaf; Chronic pancreatitis; Fibrosis; Oxidative stress

Figure

• 

  Fig. 1 (A) A histogram showing a significant decrease in the body weight in the CP group compared to the control group (a)P<0.001). The body weight was significantly improved in the CP group treated with OLE compared to the untreated CP group (b)P<0.001). (B) A histogram revealing a significant upregulation in blood glucose level, serum insulin level, and HOMA-IR score in the CP group compared to the control group (a)P<0.001). These values were significantly decreased in the CP group treated with OLE compared to the untreated CP group (b)P<0.001). CP, chronic pancreatitis; OLE, olive leaf extract; HOMA-IR, homeostatic model assessment of insulin resistance.

• 

  Fig. 2 (A, B) Histograms showing a significant decrease in serum amylase and lipase levels in the CP group compared to the control group (a)P<0.001). These values were significantly improved in the CP group treated with OLE compared to the untreated CP group (b)P<0.001). (C, D) Histograms revealing a significant increase in MDA level and decrease in SOD level in the CP group compared to the control group (a)P<0.001). These values were significantly attenuated in the CP group treated with OLE (b)P<0.001) compared to the untreated CP group. CP, chronic pancreatitis; OLE, olive leaf extract; SOD, superoxide dismutase; MDA, malondialdehyde.

• 

  Fig. 3 (A–I) Representative photomicrographs of H&E-stained sections of rat pancreas in the different experimental groups (H&E ×40, scale bars=20 μm). (A–C) Control group: showing normal pancreatic acini “A” with vesicular nuclei (arrows) and normal islets of Langerhans “i”. Normal intralobular pancreatic duct “D” is seen. (D–F) CP group exhibiting marked vacuolations “V” and pyknotic nuclei (arrowhead) of the islets of Langerhans “i” and disorganized pancreatic acini (asterisk). Congested blood vessel “BV”, inflammatory infiltrate (thick arrow), thick walled (curved arrow) intralobular pancreatic ducts “D”, decreased zymogen granules (asterisk) and fatty degeneration “F” are seen. (G–I) CP group treated with OLE revealing regaining of nearly normal β-cells (arrowhead), but pyknotic nuclei (arrow), islets of Langerhans “i”, and some vacuoles (curved arrow) can be also seen. Restored pancreatic acini “A” with many vesicular nuclei (arrow) except for some pyknotic nuclei (curved arrow) are seen; Restored intralobular pancreatic duct “D” is also noted. CP, chronic pancreatitis; OLE, olive leaf extract.

• 

  Fig. 4 A histogram showing a significant increase in the scores of areas of lesions, fat infiltration, and inflammation in the CP group compared to the control group (a)P<0.001). These scores were significantly improved in the CP group treated with OLE compared to the untreated CP group (b)P<0.001). CP, chronic pancreatitis; OLE, olive leaf extract.

• 

  Fig. 5 (A–F) Representative photomicrographs of Mallory trichrome stained sections of rat pancreas in the different experimental groups (×40, scale bars=20 μm). (A, B) Control group showing fine collagen fiber deposition (arrow) in between the pancreatic acini and around the normal pancreatic duct (thick arrow). (C, D) CP group showing massive collagen fiber deposition (arrow) within islets of Langerhans “i” and around the pancreatic duct (curved arrow). (E, F) CP group treated with OLE revealing moderate amount of collagen deposition (arrow) within islets of Langerhans and around the pancreatic duct (thick arrow). (G) A histogram revealing a significant increase in the area % of collagen fiber deposition in the CP group compared to the control group (a)P<0.001). This area % was significantly decreased in the CP group treated with OLE (b)P<0.001) compared to the untreated CP group. CP, chronic pancreatitis; OLE, olive leaf extract.

• 

  Fig. 6 Representative photomicrographs of immunostained pancreatic sections from the different studied groups showing increase in caspase-3 (A–C), IL-6 (E–G), α-SMA (I–K), and TGF-β (M–O) positive immunoreactions in the CP group compared to the control group (arrow). CP group treated with OLE reveals decrease in caspase-3, IL-6, α-SMA, and TGF-β immunoreaction compared to the untreated CP group (×40, scale bars=20 µm). (D, H, L, P) Histograms showing a significant increase in caspase-3, IL-6, α-SMA, and TGF-β positive immunoreactions in the CP group compared to the control group (a)P<0.001). CP group treated with OLE exhibits decrease in caspase-3, IL-6, α-SMA, and TGF-β immunoreactions compared to the untreated CP group (b)P<0.001). CP, chronic pancreatitis; OLE, olive leaf extract; IL, interleukin; TGF, transforming growth factor; SMA, smooth muscle actin.

Reference

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