Investigation on changes in the erythrocyte Malondialdehyde value in patients with colon cancer after radical surgery

To evaluate the change of erythrocytes malondialdehyde in patients with colon cancer before and after radical surgery and find out the relationship between the changes of erythrocyte malondialdehyde in peripheral veins after surgery and some surgical characteristics. Subjects and methods: 60 patients with color cancer treated with radical surgery from March 2015 to January 2017 at 103 Military Hospital were included in the research. The characteristics of the patients and the resection of the colon tumor were recorded, the oxidative stress was determined by measurement of erythrocyte malondialdehyde, blood samples were collected from the peripheral veins at 4 times: before surgery, 1 day after surgery, 3 days after surgery and 7 days after surgery. Results: 1 day after surgery, erythrocyte malondialdehyde increased compared to preoperation (p < 0.05). 3 days and day 7 after surgery, erythrocyte malondialdehyde decreased compared to 1 day after surgery but compared to preoperation, the change was not significant with p > 0.05. Conclusion: Compared to preoperation, the peripheral erythrocyte malondialdehyde increased significantly at the point of 1 day after surgery, and over time the erythrocyte malondialdehyde index tended to decrease.

INVESTIGATION ON CHANGES IN THE ERYTHROCYTE MALONDIALDEHYDE VALUE IN PATIENTS WITH

COLON CANCER AFTER RADICAL SURGERY

Pham Manh Cuong 1 ; Nguyen Van Xuyen 1 ; Trinh Hong Thai 2

SUMMARY

Objectives: To evaluate the change of erythrocytes malondialdehyde in patients with colon cancer before and after radical surgery and find out the relationship between the changes of erythrocyte malondialdehyde in peripheral veins after surgery and some surgical characteristics Subjects and methods: 60 patients with color cancer treated with radical surgery from March

2015 to January 2017 at 103 Military Hospital were included in the research The characteristics

of the patients and the resection of the colon tumor were recorded, the oxidative stress was determined by measurement of erythrocyte malondialdehyde, blood samples were collected from the peripheral veins at 4 times: before surgery, 1 day after surgery, 3 days after surgery and 7 days after surgery Results: 1 day after surgery, erythrocyte malondialdehyde increased compared to preoperation (p < 0.05) 3 days and day 7 after surgery, erythrocyte malondialdehyde decreased compared to 1 day after surgery but compared to preoperation, the change was not significant with p > 0.05 Conclusion: Compared to preoperation, the peripheral erythrocyte malondialdehyde increased significantly at the point of 1 day after surgery, and over time the erythrocyte malondialdehyde index tended to decrease

* Keywords: Colon cancer; Oxidative stress; Erythrocyte malondialdehyde

INTRODUCTION

Colon cancer is one of the most common

cancers with high morbidity and mortality

rates in many countries around the world

In Vietnam in 2010, along with rectal

cancer, colon cancer ranked fourth after

lung, stomach and liver cancer in men,

ranked fifth after breast, stomach, lung

and cervical cancers in women [1] In the

body, there is always a balance between

the production of oxidants and their

elimination with antioxidants, when this

process is imbalanced by free radicals that exceed the antioxidant capacity, may lead to oxidative stress Consequently, biological molecules such as DNA, proteins, lipids are constantly being attacked by free radicals that cause defects in biological molecules, damage to cells and tissues, and may in particular cause alteration gene Oxidative stress has also been identified in relation to the initiation and progression of many different cancers,

including colon cancer [2, 3]

1 103 Military Hospital

2 Hanoi University of Science, Vietnam National University

Corresponding author: Pham Manh Cuong (famcuong103@gmail.com)

Date received: 20/12/2018

Date accepted: 23/01/2019

Radical surgery is still the main method

for treating colon cancer, but the trauma

in surgery also causes oxidative stress for

the body due to the release of many free

radicals The research on changes in

postoperative oxidative stress has also

been investigated in many different

surgeries and in surgical treatment of

colon cancer [4, 5]

In Vietnam, the assessment of

oxidative stress has been carried out on

many different diseases, but no studies

have been done for colon cancer alone

Assessing the status of oxidative stress

through the determination of malondialdehyde

(MDA) in blood, a product of lipid

peroxidation, is widely and popularly

applied in many studies Therefore, the

aim of this research is: To evaluate the

change of erythrocyte MDA in peripheral

vein in patients with colon cancer after

radical surgery, to find out the relationship

between the changes of erythrocyte MDA

after surgery according to a number of

surgical characteristics

SUBJECTS AND METHODS

1 Subjects

60 colon cancer patients were treated

by radical surgery at Department of

Abdominal Surgery, 103 Military Hospital

from March 2015 to January 2017

* Inclusive criteria: Results of postoperative

pathology as adenocarcinoma of colon,

conducted radical surgery (R0) in accordance

with the prescribed procedure and has a

full quantitative result of erythrocyte MDA

before and after surgery

* Exclusive criteria: Body mass index

(BMI) ≥ 30 kg/m2, smoking, alcoholism,

systemic inflammatory disease, diabetes, cardiovascular disease, autoimmune diseases, patients with complications such as obstruction, perforation colon or distant metastasis

2 Method

* Research design: A cross-descriptive

research

* Research content:

The research group was divided into small groups, including the following criteria: age (< 60; ≥ 60); gender (male; female); disease stage (according to TNM I; II; III), surgical method (laparoscopic surgery; open surgery), surgical level (expanded; not expanded)

The extent of surgery that has been expanded is defined as being in addition

to colectomy, the patients were performed other combined surgeries such as gastrectomy, resection of small intestine, oophorectomy, etc or must proceed to total colectomy

Indicators of oxidative stress were determined by quantifying erythrocyte MDA at 4 times: before surgery, 1 day,

3 days and 7 days after surgery

* Surgical procedure:

Before surgery, all patients were prepared to clean the colon and taken the Fleet Phospho Soda manufactured by C.B Fleet Company Inc (USA) in the morning before surgery All patients were induced with propofol (3 - 4 mg/kg), fentanyl (1.51 g/kg) After endotracheal intubation, the patients were provided with mechanical ventilation using an air mixture containing 30% O2 and maintained anesthesia with sevoflurane (1 - 2%)

For patients undergoing laparoscopic

surgery, CO2 is pumped into the abdomen

and maintains a pressure of 10 - 12 mmHg

during the surgery, the tumor-containing

specimen is removed from the abdominal

wall through a small incision that is

approximately 5 cm long For

open-surgery patients, incision is made about

20 - 25 cm long in the middle abdominal

wall Postoperative treatment is administered

antibiotics combination of 3rd generation

cephalosporine and MDA for 7 days,

intravenous nutrition depending on the

patient’s ability to recover digestion

* Process of quantifying erythrocyte MDA:

Blood samples were taken from the

peripheral vein and inserted into the tube

containing ethylene diamine tetraacetic

acid (EDTA), the blood after taking was

stored at 4oC then centrifuged 4,000 rpm

to separate erythrocyte and plasma

Quantification of erythrocyte MDA based

on the procedure was modified from the

report of Ohkawa H et al in 1979 [6] The erythrocyte after being collected in the above step will be washed with PBS (phosphate buffer saline), then broken

to collect erythrocyte membranes Determination of MDA content by TBA test (based on the reaction between MDA and thiobarbituric acid - TBA), the absorbance

of the product is measured at a wavelength of 532 nm Total protein content determined by Bradford method, erythrocyte MDA concentration was calculated and expressed in ng/mg protein

* Statistical analysis:

The results for erythrocyte MDA are presented as mean value ± SD, for other characteristics expressed in proportional form Statistical analysis was performed

by Mann-Whitney, Friedman or Wilcoxon tests, with a p < 0.05 value considered statistically significant Data are processed and analyzed by SPSS 20.0 software

RESULTS

1 Characteristics of the research group

Table 1: Characteristics of clinical pathology and surgical treatment

Age

Gender

Disease stage

Surgical endoscopic method

Surgical level

The average age of the research group was 60.4 ± 11.3 (the lowest was 29 years,

the highest was 87 years), 25 patients were < 60 years of age; 35 patients were

≥ 60 years old Males: 36 patients and females: 24 patients There were 7 patients in

stage I (11.7%), 27 patients in stage II (45%), 26 patients in stage III (43.3%)

34 patients underwent laparoscopic surgery and 26 patients underwent open surgery

In 11 cases of surgery expansion, there were 2 patients with gastrectomy, 2 patients

with small intestine resection, 1 patient with hysterectomy, 1 patient with oophorectomy

and the rest had to with total colectomy

2 Changes of erythrocyte MDA index before and after surgery

Results of changes in erythrocyte MDA index before and after surgery are presented

in table 2, table 3 and figure 1

Table 2: Erythrocyte MDA content at postoperative time

(* Friedman test)

Table 3: p value when comparing MDA content by time points

(* Wilcoxon test)

7 ngày

3 ngày

1 ngày Trư? c m?

0,8

0,7

0,6

0,5

0,4

0,3

0,2

0,1

0,0

Figure 1: MDA variation of study group according to the time of surgery

The erythrocyte MDA after 1 day had the highest value, before surgery it had the smallest value, the difference between the 4 times was statistically significant with p < 0.05 Comparing paired difference the erythrocyte MDA values by each time showed the between MDA value before surgery with 1 day, between 1 day after surgery with after surgery 3 days and 7 days, the difference was statistically significant with p < 0.05 The difference of erythrocyte MDA between 3 times: before surgery, 3 days and 7 days after surgery had no statistical significance with p > 0.05

3 The change in erythrocyte MDA after surgery according to surgical characteristics

Table 4: Erythrocyte MDA values at different times by surgery

Surgery method

p*

(* Mann-Whitney test; ** Friedman test)

The erythrocyte MDA value in both laparoscopic and open groups according to the time after surgery, the difference was statistically significant with p < 0.05 However, from time to time, after surgery 7 days, erythrocyte MDA in open surgical group tends

to be lower than in laparoscopic group

Table 5: Erythrocyte MDA content at the time of surgery

Surgical level

p*

(* Mann-Whitney test; ** Friedman test)

Comparing the postoperative period, in the not expanded surgery group, the MDA change had statistical significance with p < 0.05; in the expanded surgery group, the MDA change did not have statistical significance with p > 0.05 Thus, after the surgery,

in the expanded surgical group, the MDA value change occurred less than compared to the not expanded surgery

DISCUSSION

In colon cancer, the higher oxidative

stress status in cancer tissue has been

confirmed by many researches [7] So surgery

for cancer treatment is a complicated

process because although surgery can

remove a large part of the oxidative mass

of the tumor, surgical injuries can stimulate a

stressful response, leading to a release of

a large amount of free radicals, disrupting

the internal balance of oxidation - antioxidant

systems So after radical surgery of colon

cancer, oxidative stress changes may

increase or decrease

Our research results evaluated the

oxidative stress through quantitative

erythrocyte MDA in peripheral blood

showed that after 1 day’s surgery, MDA

value increased significantly compared to

preoperation (p < 0.05, table 2, table 3 and chart 1) Our results as well as many other researches showed a higher level of immediate oxidative stress after surgery, because any form of injury, including surgery, leads to oxidative stress In abdominal surgery, increased formation of free radicals was also the result of ischemia - reperfusion of abdominal organs and inflammation of the body after surgery [8] The results when quantifying erythrocyte MDA at 3 days and 7 days after surgery showed that the level of oxidative stress compared with 1 day after surgery showed signs of reducing (p < 0.05), but compared with preoperation, the difference was not statistically significant (p > 0.05) This showed that after surgery, the body activates the anti-oxidation mechanism and over time, the redox-reduction

equilibrium tended to be reestablished,

but for some patients, 7 days after surgery

may not be enough for the body recovery

to fight against oxidative stress caused

by surgery

In laparoscopic surgery, the injection of

CO2 gas to maintain abdominal pressure

during surgery can increase the formation

of free oxygen radicals by inhibiting the

activity of SOD, an antioxidant enzyme

and increase the level of MDA, a product

of lipid peroxidation process [9], but

laparoscopic surgery is also known to

cause less surgery injury and inflammatory

reactions than open surgery, so oxidative

stress was lower Some research results

on colon surgery also showed that some

signs of oxidative stress were significantly

reduced in the endoscopic group,

compared to the open surgical group

[4, 10] Similarly, with the group of

patients with expanded surgery, there

often was a higher level of surgery, more

removed organs, more surgical seizures,

and the increase in oxidative stress is

also larger However, in our study when

comparing at the postoperative time

points of changes in erythrocyte MDA

between groups of laparoscopic surgery

and open surgery; between groups of

expanded and not-expanded surgery, the

results were incompatible, in the group of

laparoscopic surgery and not-expanded

surgery, postoperative oxidative stress

tended to more highly increase

CONCLUSION

In patients with colon cancer who

underwent radical surgery, compared to

preoperation, the erythrocyte MDA was

significantly increased at 1 day after

surgery, and on the 7th day after surgery, the erythrocyte MDA tended to decrease

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