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Effectiveness of Pectin on Serum Pancreatic Enzymes Activity of Type 2 Diabetes Mellitus

Background: Diabetes mellitus (DM) is a series of metabolic conditions that lead to hyperglycemia which is caused by impaired insulin secretion, poor insulin efficiency, or both. Type 2 diabetes mellitus (T2DM) is an islet paracrinopathy, in which the equal relationship between both the glucagon secreting alpha cell and the insulin-secreting beta cell is disrupted, resulting in hyperglucagonemia and hyperglycemia. Patients with T2DM are mostly characterized by being obese or having a higher body fat percentage, distributed pre-dominantly in the abdominal region. In this condition, adipose tissue promotes insulin resistance through various inflammatory mechanisms, including increased free fatty acid (FFA) release and adipokine deregulation. Pectin is a biocompatible polysaccharide polymer with intrinsic biological activity, which may exhibit different structures depending on its source or extraction method. Pectin is susceptible to physical, chemical, and/or enzymatic changes. Apple pomace and a smaller extent is the main source of commercial pectin. Pectin-modified nanomaterials has an antidiabetic agent in (T2DM) through decreasing blood glucose levels and/or insulin secretion after a sugar load, helps to reduce blood cholesterol, and reduces the risk of cardiovascular disease. Due to its high availability and low production cost, pectin can benefit treatments for weight reduction and obesity. Besides, as pectin is a weak acid, it resists dissociation in the gastric environment and binds covalently to the active sites of pancreatic lipase. Furthermore, it is highly desirable to study the effect of pectin-modified nanoparticles on the pancreatic enzymes in sera of (T2DM) patients by evaluating the lipase and amylase activities in the absence and presence of nanoparticles, evaluating of lipid profile, as well as ghrelin levels in sera of (T2DM) patients, and assess the relationship of pancreatic enzymes and studied biochemical parameters.
Method: It included a group of 39 patients with type 2 diabetes mellitus and 39 healthy people between the age of 35 to 60 years. Quantitative determination of α-amylase in serum was done by kinetic method (CNPG3) and the fasting serum glucose, lipase, total serum cholesterol, serum triglycerides, and HDL were determined by the colorimetric method. Additionally, LDL was calculated by the indirect method and VLDL was calculated by dividing the triglycerides concentration by 5. The HbA1c determination was based on the turbid metric inhibition immunoassay (TINIA) for hemolyzed whole blood and the measurement of serum human insulin and serum human ghrelin were done by an Enzyme-Linked Immunosorbent Assay (ELISA). The preparation of pectin nanoparticles, and calcium carbonate nanoparticles (CaCO3 NPs). As well, calcium pectin core-shell nanoparticles denoted as Ca@pectin NPs were synthesized coating of the surface of calcium carbonate nanoparticles with pectin polymer. The characterization of prepared nanomaterials was done by using field-emission scanning electron microscopy (SEM), X-ray diffraction (XRD), dynamic laser scattering (DLS), ultra violet visible spectrophotometer (UV-Vis-NIR), Zeta potential, and transmission electron microscopy (TEM).
Results: Results indicated a significant difference in FBS among groups, The mean levels of FBS for DM was (203.7 ± 72.3) mg/dL which was significantly higher than for the control group (88.6 ±7.64) mg/dL, ( p ≤ 0.001). The mean of HbA1c for DM (9.5 ±2.35) was significantly higher than for control (4.9 ±0.48), p ≤ 0.001. The mean levels of the Amylase & Lipase activity in the DM patients group were (71.4 ±29.11) and (49.7 ±14.69) (U/L) respectively, while the mean levels in the control group were (72.7 ±27.19) and (42.0 ±7.35) (U/L). The mean levels of Lipase activity were shown a significant difference in T2DM patients compared to the healthy control group, (p <0.05). Serum hormones were also increased significantly (P < 0.05), suggesting that DM patients had an obvious insulin resistance. The mean levels of the serum Insulin & Ghrelin in the DM patients group were (18.3 ±6.04) and (3.3 ±1.01) (IU/L) respectively, while the mean levels in the control group were (14.5 ±5.41) and (2.6 ±0.71) (IU/L). Lipid abnormalities in patients with diabetes often termed “diabetic dyslipidemia”, are typically characterized by high total cholesterol (T-Chol), high triglycerides (TG), low range of high-density lipoprotein cholesterol (HDL-C), and massively increased levels of small dense (VLDL and LDL) particles. The morphology and structure of the prepared nanopectin were characterized by using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The average size of Ca@pectin NPs increased after coating with pectin polymer, indicating successful coating of pectin on the surface of CaCO3 NPs. The particle sizes of the as-made nanomaterials were also further examined using dynamic laser scattering (DLS) measurements, demonstrating the successful pectin polymer coating on CaCO3 NPs and the formation of CaCO3@pectin core-shell NPs. The crystalline structure of CaCO3@pectin was further identified by the X-ray diffraction (XRD) pattern. The results showed that the CaCO3 NPs are highly crystalline while the Ca@pectin NPs are amorphous. These results clearly indicate the successful formation of Ca@pectin composite NPs. The optical absorption of CaCO3@Pectin NPs has the characteristic peak of CaCO3 which at near 400 nm is clear evidence for the formation of CaCO3@Pectin NPs. The pool activity of serum Amylase in the presence of CaCO3@Pectin NPs, calcium nanoparticle, and pectin polmer was 63.1< 82.2< 109.3 (U/L) respectively, while the pool activity of serum lipase was 12.9˃ 12.2< 20 (U/L). The spearman rank test analysis was used to analyse the response relationship between parameters. Mainly, serum FBS was positively related to the HbA1c, TG, and VLDL levels. In addition, TG levels were significantly associated with total cholesterol, VLDL, and Insulin levels. As a risk factor for dyslipidemia in (T2DM), serum LDL was highly significantly and positively related to high levels of total cholesterol (all P < 0.001). The overall relationship between the parameters and study cases are also the individual Correlation Coefficient of the Ghrelin, Lipase, and Amylase.
Conclusion: CaCO3@Pectin NPs have an inhibitory effect on the activity of the enzymes lipase and amylase, while the pectin polymer showed an activating effect on enzyme amylase. Therefore, it is possible to study the possibility of using pectin nanoparticles in various forms as a carrier of diabetes drugs according to the physiological state of the diabetic patient.