INTRODUCTION:

Preeclampsia has been suggested as a trigger for maternal endothelial activation. Its cause trophoblastic on placental of pregnancy is related with maternal's blood¹. The placental and maternal's blood triggers of PE remains to be doubted, but trophoblast debris from mother circulation was related to preeclampsia disease¹. In placenta preeclampsia, trophoblast exposure during hypoxia in vitro causes a bond with the apoptotic process associated with increased anti-apoptotic expression. The high content of terpenoids and antioxidants in placental trophoblast cells, especially in the muscular layer, reduces normal cells and narrowed arteries partially². Haramonting has high antioxidant. Extra virgin olive oil (EVOO), containing tocopherol (Vitamin E) which is proven effective in increasing plasma antioxidant activity in preeclampsia^3,4.

Rhodomyrtus tomentosa or Haramonting leaves are a traditional medicine (ancient times) as an antipyretic, anti-diarrheal, antimicrobial, diabetes and antioxidant drug in Asian countries such as Vietnam, China, and Malaysia, Thailand and Indonesia⁵. Treatment of urinary tract infections, heartburn, snake bites, diarrhea, dysentery, and to improve the immune system⁵. Haramonting contains phytochemical composition of lupeol, -amyrin, -amyrenonol, and botulin⁶ which have potential inhibitory activity against human cyclase oxidosqualene⁷. Phenolic compounds are also identified as a major component in haramonting^8,9. Research on haramonting leaves has focused on bioactive content and haramonting can be used as an antibacterial, diabetic drug, prevention of DNA damage, and antioxidant activity like Extra Virgin Olive Oil (EVOO)¹⁰. There hasn't been no research on the effects of administration of nanoherbal haramonting on placental histology.

MATERIAL AND METHODS:

Nanoherbal haramonting dan Extra Virgin Olive oil (EVOO):

Haramonting (Rhodomyrtus tomentosa) leaves originates from Labuhanbatu, Northen Sumatra, Indonesia. The procedure for making nanoherbal was done by High energy milling (HEM) tool at LIPI Jakarta^2,11. This research used EVOO comes from a supermarket in Medan as comparison because EVOO has been shown to have antioxidants.

Animal:

This study used 50 female rats that acclimatized for 2 weeks with normal feed and drinking water ad libitum. The cage was cleaned as quickly as 2 times. Feed was given in the morning the afternoon. Female rats are mated by placing 2 male rats in one cage for mating process. Copulation occurs with a vaginal or copulatoy plug that closes the cervix until the vulva on the next day. That day was the 0 th pregnancy day. This study consisted of 5 treatments in which a PE mouse model was made by injecting NaCl 6% 3ml/day/KgBW subcutaneously from pregnancy 6-12. The study design was negative control (C-): Normal pregnant rats, Positive control (C+): PE rats, P1: EVOO 1 ml/kgBW/day orally (pregnancy 13-19), T2: Nanoherbal haramonting 100mg/KgBW (pregnancy 13-19). T3: EVOO 0.5ml/KgBW/day orally and nanoherbal haramonting 50mg/KgBW (pregnancy 13-19)^2,12.

Blood Pressure and Proteunaria:

Blood pressure and proteunaria examinations were carried out at the Animal House Laboratory of the University of Sumatera Utara (USU). Blood pressure was measured using an electronic blood pressure device (noninvasive sphygmomanometer) at the base of the tail, and proteunaria values use a urine dipstick. Examination time was before injection of NaCl or normal (day 5th days of pregnancy), after injection of NaCl (preeclampsia rats or 13rd days of pregnancy) and after being given drugs or before dissected (19th days of pregnancy). Rats suspected of preeclampsia with blood pressure greater than 120/80mmHg and proteiunaria values of 30mg/dL in urine 24 hours or +1 on urine dipstick^2,12.

Placental:

The placenta was made into paraffin blocks and then stained by Hematoxylin Eosin (HE) staining. The trophoblast cells on the placenta of pregnant rats observed using a microscope at 40x magnification².

Data analysis:

Research data using SPSS software version 23 using the ANOVA test at 5% level then continued with the Post Hoc-Duncan test. If the data are not normally distributed and/ or the variance is not homogeneous, then use the Kruskal Wallis test and then proceed with the Mann-whiney test.

RESULTS:

Fig. 1: Systol Blood Pressures, C-: Normal pregnancy, C +: PE , T1: EVOO, T2: Nanoherbal haramonting, T3: EVOO and nanoherbal haramonting.

Systole blood pressure was examined in the 5th pregnancy (before injection of NaCl), 13th (Preeclampsia) and 19th (after giving EVOO and Nanoherbal haramonting). In the 5th days of pregnancy had normal blood pressure. After injection of 3ml 6% NaCl (animal model of preeclampsia) the blood pressure of systol rats increased (P <0.05). On the 19th days of pregnancy after administration of the two herbs, the blood pressure of systol rats decreased in the treatment group (P <0.05). The lowest blood pressure reduction starts from the T3, T2 and T1 groups (Fig. 1). From these data NaCl can increase blood pressure or animal models of preeclampsia and both herbs can reduce rats blood pressure.

Fig.2: Blood Pressures Diastol, C-: Normal pregnancy, C +: PE, T1: EVOO, T2: Nanoherbal haramonting, T3: EVOO and nanoherbal haramonting.

Diastolic blood pressure (Fig. 2) was also examined in the 5th pregnancy (red graph), 13rd pregnancy (yellow graph) and 19th pregnancy (green chart). Rats had normal diastolic blood pressure on the 5th day of pregnancy. After injection of 3ml of NaCl 6% (rats model of preeclampsia) the blood pressure of diastolic rats increased but not as high as systol blood pressure. After administration of the two herbs, the diastolic blood pressure was decreased on the 19th day of pregnancy (P <0.05). The lowest drop on diastolic blood pressure starts in the T1, T3 and T2 groups (Fig. 2). From these data NaCl can increase diastolic blood pressure and to making of animal models of preeclampsia and haramonting and EVOO can reduce rat diastolic blood pressure.

Fig.3: Proteunaria Value, C-: Normal pregnancy, C +: PE, T1: EVOO, T2: Nanoherbal haramonting, T3: EVOO and nanoherbal haramonting

Proteunaria values (Fig. 3) were also examined in the 5,13th and 19th days of pregnancies in the morning. Pregnant rats had normal proteunaria values on the 5th pregnancy. After injection of 3ml of NaCl 6% (rats model preeclampsia) the value of proteunaria increased (P<0.05) then administration of nanoherbal haramonting and EVOO was decreased proteunaria values on the 19th days of pregnancy. However, there was no significant difference in each treatment (Fig. 3) So it was difficult to determine which treatments from herbs were more effective but both of these plants have the same benefits.

Fig.4: Normal Trophoblast, C-: Normal pregnancy, C +: PE, T1: EVOO, T2: Nanoherbal haramonting, T3: EVOO and nanoherbal haramonting.

Fig.5 : Histology of Placental C-: Normal pregnancy, C +: PE, T1: EVOO, T2: Nanoherbal haramonting, T3: EVOO and nanoherbal haramonting. a. Tropbolast, b. Blood vessel

Trophoblast plays a role in the process of formation of the placenta. The lowest of normal trophoblast number was found in placenta preeclampsia (C +) and the highest was the combination of EVOO and nanoherbal haramonting (T3) (Fig. 4) (P <0.05). The trophoblast was very large and blood vessels are still thick on normal placental but C + (PE) there was necrosis in the placenta, cells was empty, the relationship between cells was far apart and thinning of the arteries. In T1 with EVOO, the form of dense cells and blood vessels was also thick as the control group. In T2 with nanoherbal haramonting there were necrosis cells but only a few and blood vessels were still thick. In T3 with the administration of EVOO and nanoherbal haramonting there was a large number of trophoblasts and blood vessels that were slightly thinner (Fig. 5).

DISCUSSION:

Abnormal development of placental blood vessels for placental insufficiency can cause a decrease in nutrient exchange between the mother and the fetal circulation. Such changes can adversely affect the condition of the uterus which causes various pregnancy complications such us pregnancy hypertension, premature birth, intrauterine growth restriction, stillbirth, miscarriage and preeclampsia for maternal^13-15. Cell death and disorders of the placenta and fetus cause the release of proinflammatory signals (inflammation). Reactive oxygen species (ROS) in the body, usually produced in vascular endothelial of cells trophoblast and play an important role in embryonic development, angiogenesis, increased inflammation and fetal growth¹⁵. The placenta in pregnancy really needs compounds that can increase spiral arterial remodeling, increase placental perfusion and trophoblast invasion. The compounds needed are antioxidants because the antioxidant system in the placenta can reduce the level of oxidative stress, especially preeclampsia dieases^16.

Disturbed of trophoblastic cell invasion can interfere with fetal growth. The placenta plays an important role in reducing uteroplacental perfusion and pathogenesis of PE. then causes the development of an abnormal invasion of cytotrophoblasts, and triggers the spiral cascade arterioles. which can cause interference with the mother. Placental hypoxia can cause villi and trophoblast terminal parts to be irregular and accumulate, it can cause synchronous node formation. Jam that placental ischemia also causes the release of placental factors as antiangiogenic or proinflammatory^2,17. Haramonting can repair trophoblast damage to the placenta. That is because haramonting has extraordinary antioxidant activity on radical activity, inhibits lipid peroxidation activity and decreases MDA levels¹⁸. Besides having high antioxidants, Saponin in haramonting is a secondary metabolite compound that is suspected of causing damage to cells. Saponins are natural glycosides that can cause hemolysis by affecting the action of lipid bilayer so that the formation of pores on the red blood cell membrane in the body^19,20. It is very dangerous during the placenta and fetus during pregnancy. therefore needed an effective dose in consuming this plant.

CONCLUSION:

Preeclampsia cause interference on the development of the placenta, blood vessels and decrease in the exchange of nutrients between circulation of the maternal and the fetal. The administration of nanoherbal haramonting can improve normal trophoblast and blood vessel in placenta preeclampsia (P <0.05).

ACKNOWLEDGEMENT:

We are grateful to Kemenristekdikti have funded our research (DRPM) Number: 11/E1/KP.PTNBH/2019 (Dissertation Research Grant).

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 04.11.2019 Modified on 21.01.2020

Research J. Pharm. and Tech. 2020; 13(8):3879-3882.

DOI: 10.5958/0974-360X.2020.00686.1