THE EFFECT OF HALOTHANE INHALATIONAL ANAESTHETICS ON LIVER CELLS(Experimental Study of Halothane Toxic Effect in BALB C Mice) - Diponegoro University | Institutional Repository (UNDIP-IR)

CHAPTER 5
RESULT

5.1 RESULT OF COUNTING TISSUE CHANGES FROM HE STAINING
EXAMINATION
Tissue changes were observed from HE staining examination with 1000x
magnification in 10 fields from randomized choosing. The examiner counted the
number of cells with nucleus changes (i.e, enlarge, karyorhexis, and karyolysis).
The results were expressed in percentage of abnormal cells per all cells counted
on those fields. From total 30 mice, 30 livers of the mice were made into tissue
slides.

37

a

A

c

b

c
b
B
Figure 6. HE staining examination using 1000X magnification from: A. control
group. The mean of abnormal cells percentage was 17.65%. a: enlarge nucleus;
B. group 3. b: karyorhexis; c: karyolisis.

38

Table 4. Percentage of cell changes from HE staining examination
Groups
Control
Group 1
Group 2
Group 3
Group 4

Mean (%)
17.65

40.93
51.94
61.22
60.77

Median (%)
14.35
40.39
52.17
60.79
63.39

SD
5.14
1.71
1.21
0.59
3.60

As shown in Table 4, control group has the smallest percentage of nucleus

changes (mean: 17.65%) and group 3 has the highest percentage of nucleus
changes (mean: 61.22). Figure 6 shows that box plot median percentage of
nucleus liver changes in group 4 is higher than other groups.

80.00

60.00

18

40.00

20.00

0.00
control group

group 1 2w

group 2 4m

group 3 6w

group 4 8w

Groups

Figure 7. Box plot median percentage
changes in control group, group 1, group 2, group 3 and group 4.

The significance of these differences was revealed using statistic test.
Using the Shapiro-Wilk test (test full result was attached on the appendix), we
found that the data had normal distribution which can be seen in appendix 2, yet

39

the data variant was not equal, so we could not perform One Way Anova test to
know the comparison among all groups. We performed Kruskal-wallis test and the
result was statistically significant (p = 0.00). Subsequently, we carried out Mannwhitney test as Post Hoc test to know the statistic comparison between each
group.

In figure 7, we can see that the percentages of total abnormal cells were
positively correlated with the time of exposure, thus, also with the dose of
halothane. Interestingly, in the group 4, after six weeks halothane exposure
followed by two weeks of free halothane, the mean of abnormal cells were less
than group 3 which exposed by halothane for six weeks.

However, overall

abnormal cells were higher in group 4 which suggest the cells damage still
continued regardless the cessation of halothane exposure. The comparison
between these two groups was not statistically significant. Thus, I suggest that the
liver damage may be irreversible.

Table 5. Comparison between each group (Post Hoc test). It is considered as
statistically significant if P