Int J Stem Cells.  2017 Nov;10(2):129-143. 10.15283/ijsc17035.

Combination of Obestatin and Bone Marrow Mesenchymal Stem Cells Prevents Aggravation of Endocrine Pancreatic Damage in Type II Diabetic Rats

• Hussien NI
• Ebrahim N
• Mohammed O
• Sabry D

• KMID: 2400867
• DOI: http://doi.org/10.15283/ijsc17035

Abstract

One of the new promising therapies in treatment of diabetes mellitus is mesenchymal stem cells (MSCs) which have an interesting therapeutic potentiality based on their paracrine effect and transdifferentiation potentiality. Also obestatin improves the generation of functional β cells/islet-like cell clusters in vitro, suggesting implications for cell-based replacement therapy in diabetes. So the aim of this study was to evaluate the effect of combination of both MSCs and obestatin on an experimental model of type II diabetes mellitus (T2DM). Sixty male rats were divided into; group I (control group), group II (T2DM group) induced by administration of high fat diet (HFD) and injection of streptozotocin (STZ) in low dose, group III (T2DM treated with MSCs), group IV (T2DM treated with obestatin), group V (T2DM treated with MSCs and obestatin). Fasting blood glucose, C-peptide, insulin and lipid profile were measured. HOMA-IR and HOMA-β were calculated. Pancreatic expression of insulin, glucagon like peptide -1 (GLP-1) and pancreatic duodenal homeobox 1 (Pdx1) mRNA levels were measured. In addition pancreatic histological changes, insulin and Bax were analyzed by immunohistochemical examination of islets of Langerhans. Diabetic rats showed significant increase in HOMA-IR, serum glucose and lipid profile levels with significant decrease in insulin, HOMA-β, GLP-1 and Pdx1 levels. MSCs and obestatin caused significant improvement in all parameters with more significant improvement in combined therapy. The protective effects afforded by MSCs and obestatin may derive from improvement of the metabolic profile, antiapoptosis and by increase in pancreatic GLP-1and Pdx1 gene expression.

Keyword

Mesenchymal stem cells; Obestatin; Type II diabetes mellitus

MeSH Terms

Animals
Blood Glucose
Bone Marrow*
C-Peptide
Diabetes Mellitus
Diet, High-Fat
Fasting
Gene Expression
Genes, Homeobox
Ghrelin*
Glucagon
Glucagon-Like Peptide 1
Humans
In Vitro Techniques
Insulin
Islets of Langerhans
Male
Mesenchymal Stromal Cells*
Metabolome
Models, Theoretical
Rats*
RNA, Messenger
Streptozocin
Blood Glucose
C-Peptide
Ghrelin
Glucagon
Glucagon-Like Peptide 1
Insulin
RNA, Messenger
Streptozocin

Figure

• 

  Fig. 1 Homing of MSCs in pancreas. (A) Primary culture of MSCs showing many spindle-shaped stem cells (white arrows) ×200. (B) Fluorescent microscopic image of a section in pancreas of rat in group III demonstrating the green fluorescence of MSCs labeled with GFP two week after implantation (white arrows) ×200. (C) Flow cytometric chart analysis for surface antigens of MSCs. They were positive for CD29 and CD90.

• 

  Fig. 2 Photomicrographs sections (immunohistochemistry) from rat tail pancreas showing. (A) GFP expression in transplanted MSCs. (B) CD105 expression in transplanted MSCs.

• 

  Fig. 3 Photomicrographs sections (H&E) from rat tail pancreas showing. (A) A section from control rat illustrating pancreatic lobules (bent arrows) separated by thin connective tissue septae (black arrow heads), islet of Langerhans (white arrow head), blood vessels (white ↑) and pancreatic ducts (black ↑)×100. (B) A section from a control rat revealed an islet of Langerhans (black arrow head) containing β cells in the center (white arrow head) and most probably α cells in the periphery (bent arrow), blood vessel (black ↑) and pancreatic acini formed of pyramidal cells with basal basophilia and apical acidophilia (white↑)×400. (C) A section of a rat from group II revealed two shrunken distorted islets of Langerhans with marked loss of their cells (arrow head), some cells with vacuolated cytoplasm (↑) and darkly stained nuclei. Note distorted pancreatic acini (bent arrow)×400. (D) A section of a rat from group III showing an islet of Langerhans (arrow head) with vacuolated cells (↑) surrounded by exocrine pancreatic acini ×400. (E) A section of a rat from group IV showing an islet of Langerhans (arrow head) and some cells with vacuolated cytoplasm (bent arrows)×400. (F) A section of a rat from group V revealed well-defined islet of Langerhans (arrow head) composed of cords of endocrine cells which are small with a pale-stained granular cytoplasm, exocrine pancreatic acini (bent arrows) and small blood capillaries (↑)×400.

• 

  Fig. 4 Photomicrographs sections in the tail of pancreas revealed islets of Langerhans (immunohistochemistry anti-insulin antibodies ×400) showing. (A) Strong positively stained secretory granules of the β cells in a control rat. (B) Mild positively stained secretory granules of the β cells in a rat from T2DM group. (C) Moderate positively stained secretory granules of β cells of a rat from T2DM+MSCs group. (D) Moderate positively stained secretory granules of β cells in a rat from T2DM+obestatin group. (E) A highly positive stained secretory granules of β cells in a rat from T2DM+MSCs+obestatin group.

• 

  Fig. 5 Photomicrographs of sections in the tail of pancreas revealed islets of Langerhans (immunohistochemistry anti-Bax antibodies ×400) showing. (A) Negative immunostaining of the β cells in a control rat. (B) Highly positive Bax immunostaining in the islet of Langerhans indicating increasing rate of cellular apoptosis in a rat from T2DM group. (C) Weak positive Bax immunostaining in the islet of Langerhans in a rat from T2DM+MSCs group. (D) Weak positive Bax immunostaining in the islet of Langerhans in a rat from T2DM+obestatin group. (E) Minimally expressed Bax immunostaining in the islet of Langerhans in a rat from T2DM+MSCs+obestatin group.

• 

  Fig. 6 The mean area percentages of insulin and Bax immunostaining for all groups. aSignificant difference (p<0.05) compared with control group. bSignificant difference (p<0.05) compared with T2DM group. cSignificant difference (p<0.05) compared with T2DM+MSCs group. dSignificant difference (p<0.05) compared with T2DM+obestatin group. eSignificant difference (p<0.05) compared with T2DM+MSCs+obestatin group.

Reference

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