RESULT AND DISCUSSION
HISTOLOGICAL AND HISTOMETRIC CHANGES IN UTERUS
After undergoing the histological process, the slides that have been prepared were observed using a microscope. The figure of 4x magnification is shown in Figure 4.1
Table 4.1: Effect of the treatment on histometric changes of uterus in mice.
(?m) Thickness of endometrium
(?m) Thickness of myometrium
(?m) Thickness of perimetrium
Group A 504.38 ± 64.91 250.73 ± 66.02 99.42 ± 14.27 154.21 ± 31.90
Group B 405.71 ± 89.86 213.45 ± 81.70 82.93 ± 21.34 109.28 ± 28.85
Group C 589.85 ± 59.00 321.67 ± 72.49 134.26 ± 27.00 133.98 ± 43.65
Group D 876.48 ± 232.43 485.61 ± 162.73 210.12 ± 80.74 180.74 ± 33.80
Mean ± Standard deviation
The mean of the overall uterus wall thickness, endometrium thickness and myometrium thickness are D > C > A > B meanwhile the mean of perimetrium thickness is D > A > C > B.
Upon microscopic observation, the uterus structure of nicotine-treated mice group (Group B) was having more damages comparing with the control group. Besides, the thickness of the endometrium, myometrium and perimetrium of this group are not evenly distributed compared to the control group. Their lumen also showed that it was more loosen comparing with the control group. As shown in Table 4.1, the thickness of overall uterus wall, endometrium, myometrium and perimetrium were decreased compared to the control group.
For the group that was treated with both nicotine and ?-tocotrienol (Group C), their microscopic observation shows that the uterus structures were similar with the control group. The overall thickness of uterus, endometrium and myometrium were increased compared with control group while the perimetrium thickness was decreased compared to the control group. This may be due to the anti-oxidative effect of a-tocotrienol that reversed the adverse effect of the nicotine.
When the mice were treated with ?-tocotrienol only (Group D), the structure of the uteruses was solid and rigid compared to the control group. The thickness of the layer of the uterus wall was evenly distributed. They also have the well-shaped lumen. This observation shows that ?-tocotrienol also improved the structure of normal mice. The thickness of overall uterus wall, endometrium, myometrium and perimetrium show increase in thickness.
The hypothalamus regulates the rhythmic release of pituitary gonadotrophins such as FSH, LH and prolactin through neural stimulus to GnRH (Sharp & Beyer, 1986). The orderly event of follicular growth and ovulation depends upon the pituitary FSH, LH and prolactin.
The gonadotropin secretion is controlled by central nervous system. The secretion is stoped after the vascular connection between the anterior pituitary gland and the hypothalamus is interrupted (Maurice Goodman, 2009). Investigations on nicotine indicate that nicotine being a central nervous system influencing drug inhibits the release of gonadotropins from the pituitary (Anderson, Fuxe, Eneroth, & Agnati, 1982; C. A. Blake, 1978; Charles A. Blake, 1974). As reported by Patil in 1998, the destructive effect of nicotine which it reduces the thickness of uterus wall, endometrium and myometrium compared to the control group.
When the mice are treated with ?-tocotrienol in group C and group D, the thickness of uterus wall is increased in size compared to the control group treated with corn oil. Tocotrienols exhibit antioxidant function like tocopherols, and it was discovered that tocotrienols exhibit more antioxidant activity. It was several-fold more effective than tocopherols in inhibiting the proliferation of mouse mammary tumour epithelial cells and in inducing apoptosis (McIntyre et al., 2000). On the other hand, tocotrienol administration reduces oxidative protein damage and extends the mean lifespan of C. elegans. compared to tocopherol (Adachi & Ishii, 2000).
Nicotine gives the adverse effect to the uterus while ?-tocotrienol reversed the adverse effect of the nicotine on the uterus. However, this finding is still in experimental level to verify the effectiveness of ?-tocotrienol. It is a stepping point for further experimentation in the future regarding the usage of ?-tocotrienol in various health application.