SERUM HEPATOCYTE GROWTH FACTOR IN EGYPTIAN PATIENTS WITH CHRONIC KIDNEY DISEASE

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SERUM HEPATOCYTE GROWTH FACTOR IN EGYPTIAN PATIENTS WITH CHRONIC KIDNEY DISEASE
Rasha
Darwish
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

CKD is a major health problem; its prevalence in both initial and final (renal failure) stages, and to the high cost and poor results of treatment (1). CKD is characterized by progressive loss of the renal parenchyma and the destruction of functional nephrons, which finally causes end stage kidney disease (2). Hepatocyte Growth Factor (HGF) is a heterodimer molecule mesenchymally derived, pleiotropic, multifunctional cytokine and consisting of 69 kDa α-chain and a 34 kDa β-chain (3).

HGF is produced by different tissues such as liver, kidney, lung and spleen. It exerts multiple biological activities on different cell types as in injured tissues. It helps tissue repair and regeneration rapidly as it has great role in stimulation of hepatocytes and renal cells proliferations (4).

HGF exhibits mitogenic, anti-apoptotic and morphogenic activities on renal cells and appear to stimulate renal repair in acute renal failure and it may play a role in chronic kidney disease by counteracting tissue fibrosis with elevation of serum HGF levels in patients with chronic kidney disease and it has been suggested that the level of HGF is correlated with renal functions (5).


In this study, we tried to look for the correlation between HGF and renal function in patients with chronic kidney diseases or end stage kidney disease and evaluate HGF as a marker of chronic kidney disease.

This is a cross-sectional, case-control study was conducted on 42 CKD patients and 10 age and sex matched healthy volunteers. 52 patients were divided into four groups: Group 1 (control group) including 10 patients, Group 2 including 6 patients CKD stages (I and II) with eGFR (≥60 mL/min/1.73 m2), Group 3 including 10 patients CKD stages (III and IV) with eGFR (15- 59 mL/min/1.73 m2) and Group 4 Including 26 patients (14 males and 12 females) with End Stage Kidney Disease (ESKD) patients on regular hemodialysis.

Exclusion criteria included any patient with renal transplantation, patients with malignancies, pregnant women, patients with liver diseases, patients with severe inflammatory or severe concomitant diseases and patients with lung diseases.

All participants were subjected to thorough history and comprehensive physical examination. The disease duration and medication history were recorded for all patients. A blood sample was obtained from every patient to test for renal functions (serum creatinine , serum urea, serum sodium and serum potassium), estimation of GFR by MDRD equation: MDRD equation=186 × Scr-1.154 × Age-0.203 × (1.212 if black) × (0.742 if female), liver functions (liver enzymes - serum albumin), serum hepatocyte growth factor, pelvi-abdominal ultrasonography and chest X-ray.

Human HGF Assay: Serum samples were collected from participants and stored at -70C. We used BioSource international, Inc. hHGF kits which is a solid phase sandwich Enzyme Linked Immuno-Sorbent Assay (ELISA).

Microsoft computer statistics package was used for data analysis. Data were summarized as mean and standard deviation. Comparison between groups was evaluated using Student's t-test. Comparison between more than 2 independent groups was evaluated using ANNOVA test.


Results are summarized in tables 1-7and figures 1-7.

There was no statistically significant difference across the four groups regarding sex (Table 1). (Table 3) This table shows high significant difference among different groups as regarding serum level of HGF. (Table 4) This table shows a high significant difference of group 4 from all others groups. There was as well a high significant difference of group 3 from control group and group 2, while the least significance difference was found between group 2 and control group. (Table 5) This table shows high significant difference among different groups as regarding serum albumin, eGFR, BUN and serum creatinine but no significant difference regarding serum ALT, serum AST, serum K+ and serum Na+.(Table 6) This table showed that there was a high statistically significant difference between different sonographic groups as regarding mean value of serum HGF of each group.

 

Table (1): Gender distribution among studied groups

 

Stage

χ2

P

Group1

control

Group 2

(I – II)

Group 3

(III–IV)

Conclusions

In group 2 (CKD stages I and II ), there were mild to moderate increase of HGF levels with mean value of serum HGF was 713.3 ± 123.9 ng/L while there were more increase of the levels of serum HGF in group 3 ( CKD stages III and IV) with mean value was 1039 ± 165.8 ng/L (Table 3). These results were consistent with the study of Sugimura et al. (5) who revealed that non-dialysis patients with renal insufficiency had significantly higher serum HGF than normal subjects (p < 0.001), and the elevated serum HGF correlated with their serum creatinine levels. The increase of HGF levels on hemodialysis patients was more than the patients with chronic kidney stages (I, II, III and IV) (Table 3). These findings coincide with Libetta. et al. (6) who stated that hemodialysis causes a prompt and prolonged release of HGF into the circulation, raising HGF serum concentration up to 30 times.

We found in our study, as well, that there is a prompt increase of levels of serum HGF in group 4 (CKD stage V) with mean value of 1616.5 ± 324.5 ng/L (Table 3), (Fig 2). There was as well a high significant difference of group 4 from all others groups (Table 4). This is in agreement with Liu Y et al. (7) study findings. Different mechanisms might underlie the elevated HGF levels found in HD; Lohr J et al. (8) demonstrated that dialysis treatment induces release of interleukin-1 (IL-1) and tumor necrosis factor, which may act as HGF inducers (the so-called Injurins). The release in blood of cytokines results from the fact that extracorporeal circulation is associated with leukocyte activation. Activated peripheral blood leukocytes are a possible source of HGF. In addition, dialysis may cause the release of coagulation products that convert HGF into its active form. Thus, dialysis may induce the release of HGF either from activated peripheral blood leukocytes or from tissue cells stimulated by cytokines.

In addition, Alvarez et al. (9) confirmed that the release of HGF takes place even if dialysis is performed without heparin since it peaked early after the start of extracorporeal circulation in their study. This proves that it does not depend on heparin-induced shedding of receptor bound HGF from peripheral tissues.

Therefore, at least theoretically, increased HGF activity induced by dialysis may be pathophysiologically relevant in patients requiring renal replacement therapy. On the other hand, there was also significant increase of serum HGF levels with increasing age (Table 7). This is in agreement with Mizuno et al. (10), who stated the same finding. When considering that higher age is usually accompanied by higher prevalence of comorbidities including CVD and higher circulating concentrations of several inflammation markers (11), one may speculate that inflammation could be the common pathway linking HGF with higher age. This is supported by some studies showing that inflammatory cytokine stimulation induces up-regulation of HGF.

In our study, there was significant correlation between renal sonographic appearance and the levels of serum HGF with increasing the serum HGF level with increasing the degree of nephropathy (fibrosis). The mean level of HGF in normal kidneys ultrasonography was (1004) ng/L, in the grade I nephropathy was (1233) ng/L, in the grade II nephropathy was (1464) ng/L, in the grade III nephropathy (1739) ng/L and reaching to highest level in marked fibrosis when kidneys became shrunken up to (1800) ng/L (Table 6),(fig 3).

Sonographic determination of renal length and cortical echogenicity correlates with chronic, irreversible renal disease. For each histologic measure of disease (renal fibrosis), the correlation was substantially stronger with echogenicity than with renal length, combining kidney length and echogenicity provided much better discrimination of renal fibrosis as with increasing echogenicity of kidneys in ultrasonography, the degree of renal fibrosis increased (12).

In our study we found that the most common cause for CKD was diabetes (DM) and hypertension (26.2%) followed by hypertension alone (19%) then by DM alone (9.5%) and unknown causes represented (9.5%). This coincides with Redmon et al. (13) who mentioned that the most common recognized cause of CKD is diabetes mellitus. Others included idiopathic (i.e. unknown cause), hypertension, and glomerulonephritis, causing all together about 75% of all adult cases.

In our study we found that the most common associated disease was IHD in both patients with ESKD on regular hemodialysis and patients with CKD not on regular hemodialysis. This is in agreement with the well-known fact that evens a mildly reduced renal function results in a dramatically increased risk of premature CVD (14). That is of serious clinical importance since Cardiovascular disease (CVD) is the major cause of death in patients with ESRD. Additionally, impaired cardiac function can worsen renal function, a complex interaction known as the cardio-renal syndrome (15).

The current study revealed high significant difference among different groups as regarding serum albumin (P <0.001), mean value of serum albumin in group 1 (control group) was 4.8 ± 0.4 g/dl , mean value of serum albumin in group 2 (stages I and II) was 3.6 ± 0.8 g/dl, mean value of serum albumin in group 3 (stages III and IV) was 3.6 ± 0.5 g/dl, mean value of serum albumin in group 4 (stage V) was 3.5 ± 0.3 g/dl (Table 5).These results are going in harmony with the results obtained by Kikuchi et al.(16); They measured it in 1138 patients with CKD not on hemodialysis and found that these patients had a  mild decrease of serum albumin with a  mean value of  3.8 ± 0.6. Friedman and Fadem(17) measured it in ESKD patients on regular hemodialysis and found serum albumin level was significantly lower (3.3±0.6 g/dl; P<0.001). These low albumin values may be due to CKD associated abnormalities such as malnutrition, inflammation, infection, protein loss into urine and dialysate and over hydration (17). Its clinical significance comes from the association of hypoalbuminemia with protein-energy wasting (PEW) which is progressive loss of body protein mass and energy reserves linked to higher morbidity and mortality rates. This is among numerous complications of CKD, usually in advanced stages of CKD, possibly affecting 18%–75% of patients with ESRD.

Our study showed no statistically significant correlation among the 4 groups as regarding the liver enzymes AST and ALT (Table 5).This is in agreement with Trevizli et al. (18), who mentioned that inflammatory activity of the liver was significantly lower in HD patients with no difference of liver enzymes between HD patients and control group. Even in HD patients who are infected with HCV, serum aminotransferase levels were found usually normal or only slightly elevated (19)

Our study showed that there was no statistically significant correlation found among the 4 groups as regards gender. These results agree with Libetta et al. (20), who demonstrated that there was no statistically significant difference in serum HGF across gender.

Conclusion:

From the previously mentioned results we can conclude that there was an increase in the level of serum HGF levels in patients with chronic kidney disease with prompt increase in CKD stage V, especially ESKD on regular hemodialysis. Thus, serum HGF may be a good indicator for the degree of renal fibrosis in CKD patients.



Ethical Committee Approval: The local ethical committee of the Internal Medicine department, Theodor Bilharz research Institute, Cairo, Egypt, approved this work.

Human and Animal Rights: All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent: “Informed consent was obtained from all individual participants included in the study”.

Conflict of interest: The authors have declared that no conflict of interest exists.

Funding: This research did not receive any specific grant from any funding agency in the public, commercial or not-for-profit sector.

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