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Diabetic Nephropathy

We know that Wobenzym can increase levels of C-peptide in autoimmune mediated diabetes, such as type 1 diabetes. In addition to showing much insulin the pancreas can still make, C-peptide decreases the progression so the common complications of diabetes, including the diabetic nephropathy.
We also note that systemic enzymes can decrease the formation of “advanced glycation end products” (AGEs), which are associated with diabetic nephropathy and other complications of diabetes.
IN addition, abnormal cytokines levels are normalized, including transforming growth factor beta-1 (TGF-b1), and interleukin 6.
What we have come to realize, is that the complications of diabetes are strongly mediated by the immune system. We see that controlling blood sugar with insulin is not enough to prevent complications such as diabetic nephropathy. The abnormal cytokine levels that occur with diabetes can safely and effectively be normalized with Wobenzym.
                  From FREQUENTLY ASKED QUESTIONS, with answers by Joseph J Collins, RN, ND
                  Read the special section: KIDNEY AND BLADDER CONDITIONS & WOBENZYM®

What the literature says about Systemic Enzyme Support and:

Diabetic Nephropathy

Enzyme Therapy in Diabetic Nephropathy – Experimental and First Clinical Data  

*Stauder G., *Wood G., **Paczek L. Enzyme Therapy in Diabetic Nephropathy – Experimental and First Clinical Data. 6th Taormina Course of Nephrology. October 20th - 22th, 2000, pp. 227-232, Editoriale Bios 2000 PZ 22 (5-14-3)-(19-11-3) [Czech summary] * Mucos Pharma, Geretsried, Germany. ** Transplantation Institute, University Warsaw, Poland  

English summary of lecture
Diabetic nephropathy is characterized by cell hypertrophy, thickening of the basement membranes and accumulation of extracellular matrix (ECM), that is attributed to an elevated protein synthesis and an inhibition of protein degradation, the latter due to reduced proteolytic activity.
The main trigger for this process is overexpression of transforming growth factor beta-1 (TGF-b1). This is induced by numerous factors such as hyperglycemia, stimulation of the RAS, formation of advanced glycation endproducts (AGE), elevated IL-6 levels, and increased mesangial stretch. A reduction of TGF-b1 levels was documented to be associated with a retardation of disease progression. Based on the findings in endothelial cells that the receptor for AGEs (RAGE) is trypsin-sensitive, the modulatory action of this serine protease was investigated in tubule cells. The distinct overexpression of TGF-b1 as well as the hypertrophy of the cells, induced by AGE-BSA, were normalized after coincubation with trypsin. In addition, the cellular accumulation of AGEs was markedly reduced. The enzyme therapy (12 mg/day of a mixture of the active ingredients of Phlogenzym®) was able to reduce the increased intraglomerular TGF-b content in rats. In a second study in uninephrectomized, STZ induced diabetes in rats the combination of Phlogenzym® with the ACE inhibitor enalapril showed an almost 67% reduction of glomerular sclerosis, while single treatment with either enalapril or Phlogenzym® led to a 20-30% reduction only, indicating clear additive effect.
In human studies (patients with rheumatoid arthritis or myelofibrosis), elevated serum levels of TGF-b&#61489; were diminished by oral enzyme therapy. A clinical pilot study in patients with diabetic nephropathy demonstrated that oral enzyme therapy (2 tablets t.i.d.) is able to reduce enhanced levels of IL-6 both in serum and urine.A clinical double-blind placebo controlled pilot study on 24 patients with diabetic nephropathy, stages III or IV, was performed in 4 centers in Germany and Poland. Either the enzyme preparation Phlogenzym® or placebo was administered double-blinded for 16 weeks. 21 patients, mean age 51.3, and 53.5 years, respectively, were evaluated. Five patients in the enzyme group were suffering from diabetes type I, 5 patients from type II; in the placebo group 5 patients were suffering from the type I and 6 patients from type II. Five patients (enzyme group) had stage III nephropathy (microalbuminuria), 5 patients stage IV (macroalbuminuria); in the placebo group 4 patients had stage III, and 7 patients stage IV. At baseline, 7 patients in the enzyme group had proteinuria <1 g/day, 3 patients >1 g/day; in the placebo group 7 patients had proteinuria <1 g/day, and 4 patients >1 g/day. The groups were comparable. Blood glucose and mean blood pressure were controlled effectively.
At baseline, a proteinuria (median) of 0.4 g/day was measured in the enzyme group, 0.8 g/day in the placebo group. After 16 weeks the value was unchanged in the enzyme group (0.36 g/day), whereas it slightly deteriorated to 1.08 g/day in the placebo group (p > 0.05).
The albuminuria tended to lower levels (from 242.5 mg/day to 200.0 mg/day) in the enzyme group; in the placebo group, a slight increase from 508.0 mg/day to 562.0 mg/day was observed (p > 0.05). Creatinine clearance did not change in either group during the 16-week treatment period (which was not at all expected in this short time), while serum creatinine tended to decline in the enzyme group (from 1.05 mg/dl at baseline to 0.95 mg/dl), and remained unchanged (at 1.2 mg/dl) in the placebo group (p = 0.0279).
There were recorded only 2 drug related side effects, 1 under placebo (mild diarrhea) and 1 (mild constipation) under enzyme therapy. Thus, the enzyme therapy proved to be safe.
Another double-blind, placebo controlled clinical trial with Phlogenzym® will be performed in 16 centers in Europe.
Poster Reference Number 35.  

Advanced glycation end products (AGEs)-induced expression of TGF-b1 is suppressed by a protease in the tubule cell line LLC-PK1 

Xiang G., Schinzel R., Simm A., Münch G., Sebekova K., Kasper M., Niwa T.,  Schmitz Ch. and Heidland1 A. Advanced glycation end products (AGEs)-induced expression of TGF-b1 is suppressed by a protease in the tubule cell line LLC-PK1. Nephrol Dial Transplant 2001, Vol. 16, pp. 1562-1569. 555 KA [Czech abstract]  

Abstract: Background. Advanced glycation end products (AGEs) are assumed to play a key role in diabetic nephropathy (DN). Since little is known about their action in tubule cells, we investigated in LLC-PK1 cells: (i) whether AGE-bovine serum albumin (AGE-BSA) affects cell proliferation and expression of transforming growth factor-b (TGF-b1); and (ii) whether the AGE-induced effects can be modulated by trypsin due to interference with its binding proteins at the cell surface.
Methods. Arrested cells were exposed to vehicle (control), AGE-BSA (19-76 mM) and BSA (38 mM) in the presence or absence of trypsin (0.625-5.0 mg/ml) (2.5 mg/ml) for 24 h. We evaluated cell proliferation by cell count and by [3H]thymidine incorporation, TGF-b1 expression by reverse transcription-polymerase chain reaction (RT-PCR), and TGF-b1 protein by ELISA. In addition, cell accumulation of AGEs was studied by immunohistochemical staining of the AGE imidazolone.
Results. AGE-BSA inhibited [3H]thymidine incorporation, lowered cell number and increased cell protein content as well as TGF-b1 mRNA and protein as compared with control and BSA. Immunohistochemical staining revealed a marked intracellular accumulation of the AGE imidazolone. Co-incubation of AGE-BSA with trypsin ameliorated the impaired thymidine incorporation, the decreased cell count and the enhanced cell protein content. TGF-b1 overexpression was normalized, while TGF-b1 protein declined insignificantly. Intracellular imidazolone accumulation was strikingly suppressed.
Conclusions. In the tubule cell line LLC-PK1, AGE-BSA exerts an antiproliferative effect, most probably due to TGF-b1 overproduction. The co-administration of trypsin abrogated this alteration, very likely as a result of an interaction with AGE-binding protein(s), which is supported by the decreased intracellular AGE accumulation. These findings may be the starting point for the development of specific proteolytic enzymes to interfere with the interaction between AGEs and their receptors/binding proteins.
Keywords: AGEs; cell proliferation; imidazolone; TGF-b1 mRNA; trypsin and tubule cells  

Advanced glycation end products impair protein turnover in LLC-PK1: Amelioration by trypsin. 

Xiang G., Schinzel R., Simm A., Sebekova K., Heidland A. Advanced glycation end products impair protein turnover in LLC-PK1: Amelioration by trypsin. Kidney International 2001, Vol. 59, Suppl. 78, pp. S-53-S-57. SO 130 (5-07-1) [Czech translation of abstract]Department of Internal Medicine, Institute of Physiological Chemistry, and Institute of Clinical Biochemistry and Pathobiochemistry, University of Würzburg, Würzburg, Germany; and Institute of Preventive and Clinical Medicine, Bratislava, Slovakia 

Background. Advanced glycation end products (AGEs) are assumed to play a key role in the pathogenesis of diabetic nephropathy (DN) and other diabetic complications. While AGEs have been shown to exert marked effects on mesangial and endothelial cells as well as on monocytes/macrophages, little is known about their effects on tubule cells. Therefore, we addressed the questions of (1) whether AGE-bovine serum albumin (AGE-BSA) impairs the protein metabolism in the tubule cells, and if so, (2) whether the AGE-induced effects are mediated via a protease sensitive mechanism.
Methods. Arrested LLC-PK1 cells were exposed to a medium containing the vehicle (control, serum free), AGE-BSA (38 mmol/L), or BSA (38 mmol/L) in the presence or absence of trypsin (2.5 mg/mL) for 24 hours. We evaluated cell number, cell size, and cell protein content, as well as protein synthesis and protein degradation.
Results. After an incubation period of 24 hours, AGE-BSA decreased the cell number to 84.5 ± 5.5% of control and 82.5 ± 5.6% of BSA-treated cells (P < 0.05). [3H]-thymidine incorporation declined to 66% of control (P < 0.05), while BSA was without any effect. The same AGE-BSA dose reduced protein degradation (P < 0.05) and stimulated total protein synthesis slightly, as determined by L-[14C]Phe incorporation into acidicinsoluble proteins. These effects resulted in a rise in cell protein content (AGE-BSA vs. control, 21.9 ± 6.7%; AGE-BSA vs. BSA, 11.1 ± 6.0%, P < 0.05) and cell volume (AGE-BSA vs. control 9.4 ± 3.2%, AGE-BSA vs. BSA 18.4 ± 3.7%, P < 0.05). Coincubation with AGE-BSA and trypsin was associated with an amelioration of all investigated parameters concerning cell number, cell proliferation, raised cell protein content, decreased protein degradation, and enhanced protein synthesis.
Conclusion. These data indicate that AGE-BSA impairs cell proliferation and protein turnover in LLC-PK1 cells with a consequent rise in cell protein. Since these alterations were abrogated by coincubation with trypsin, an interference of this serine protease with the AGE-binding proteins on cell surfaces is assumed.
Keywords: diabetic nephropathy, tubule cells, protein metabolism, cell proliferation, serine protease. 

Beneficial effect of proteases on TGF-beta production in glomeruli from streptozotocin induced diabetes mellitus in rats. 

Paczek L.1, Gaciong Z.1, Bartlomiejczyk I.1, Czyzyk A.1, Heidland A.2. Beneficial effect of proteases on TGF-beta production in glomeruli from streptozotocin induced diabetes mellitus in rats. Inter. Journal of Tissue Reactions 1997, Vol. XIX, No. 1/2, pp 93, abstract 115, ISSN 0250-0868 149K/245 (19-04-2) 1. Warsaw School of Medicine, Warsaw, Poland 2. University of Wuerzburg, Germany 

7th Interscience World Conference on Inflammation, Antirheumatics, Analgesics, Immunomodulators, May 19-21, Geneva, Switzerland -  Abstract: Diabetic nephropathy (DM) is characterised by an excessive accumulation of extracellular matrix (ECM). In turn, perturbation of the glomeruli by the accumulation of fibronectin (FN) with high biological activity may induce both proliferation of mesangial cells and expansion of mesangial matrix. Transforming growth factor beta (TGF-b) seems to be a key cytokine that inhibits and terminates tissue repair as well as the development of glomerulosclerosis within the kidney.
The purpose of this study was to assess TGF-b and FN accumulation in glomeruli obtained from streptozotocin induced diabetes mellitus in rats treated via intraperitoneal route daily for 21 days with 12 mg protease mixture (Phlogenzym®, Mucos Pharma, Germany). To prevent ketoacidosis, the rats were treated daily with subcutaneous injections of ultralente insulin in a dosage of 0.5 U. TGF-b and FN were measured with EIA.
The data indicate that in glomeruli from diabetic rats TGF-b production increased significantly and the treatment with Phlogenzym restores the production to the normal level. Increased accumulation of FN observed in diabetic glomeruli was significantly reduced after enzyme treatment.