Analysis of Cytochrome c Expression on Liver Histology of Hepatitis Rats after Administration of Tin and Olive Leaf Ethanol Extract.

<b>Background and Objective:</b> Hepatitis is a liver illness caused by a viral infection, autoimmune conditions or the use of certain medicines. In molecular hepatitis treatment, cytochrome c can be used as a potential predictor of the severity of liver impairment. In Asia, particularly in Indonesia, antioxidant-rich plants include <i>Ficus</i> <i>carica</i> and <i>Olea europaea.</i> This study aimed to see what impact cytochrome c in hepatitis after these two botanicals were administered. <b>Materials and Methods:</b> Rats were grouped as follows: Normal rats with no additions or herbs (G₀), the physiological solution group (G₁), the intravenous administration of the quercetin-copper (II) (G₂), Olive leaf extract or OLE (300 mg kg¹ b.wt.) (G₃) and Tin leaf extract or TLE (100 mg kg¹ b.wt.) (G₄). For an animal model of hepatitis, the rats were given thioacetamide 280 mg kg¹ b.wt., 8 days later. The rats were dissected and blood and liver samples were collected for enzyme and immunohistochemistry examination. <b>Results:</b> Malondialdehyde (MDA), superoxide dismutase (SOD) and cytochrome c expression levels differed significantly (p<0.05) across treatment groups in rat's models of hepatitis. Hepatocytes first displayed symptoms of lipid degradation, inflammatory and necrosis cells. When administered quercetin and the two herbs, necrosis and inflammatory cells were reduced, demonstrating that OLE and TLE can enhance liver histology and lower cytochrome c expression in a mouse model of hepatitis. <b>Conclusion:</b> Administration of Olive leaf extract (OLE) and Tin leaf extract (TLE) can improve liver histology in hepatitis model rats while decreasing cytochrome c expression, which is a mechanism for hepatocyte cell death.

Suppression of Wnt Expression by Increasing PI3K in Rats Cervical Carcinoma by Andaliman (Zanthoxylum acanthopodium).

<b>Background and Objective:</b> Cervical cancer is the leading cause of death for women in the world and Indonesia. This disease originates from a malignant tumour of squamous epithelial cells caused by infection with the Human Papilloma Virus (HPV). Antioxidants can reduce oxidative stress in and there are plants from Indonesia that have high antioxidants, namely andaliman (<i>Zanthoxylum acanthopodium</i>). This study aimed to analyze the role of andaliman on PI3K and Wnt signalling in cervical cancer histology. <b>Materials and Methods:</b> The study includes 5 treatments. The control group (K-), rats cancer model (K+), rats cancer model+the dose is 100 mg/b.wt. of ZAM (P₁), rats cancer model+the dose is 200 mg/b.wt. of ZAM (P₂) and rats cancer model+the dosage is 400 mg/b.wt. ZAM (P₃). On the 30th day after ZAM administration, the rats were dissected for the paraffin block and Wnt and PI3K immunohistochemical staining was prepared. <b>Results:</b> There was a significant difference between all groups (p<0.001) in Wnt and PI3K expression. The real role of ZAM in cervical cancer tissue was seen at the highest ZAM dose (P₃). Irregular mucosal folds and stretched interstitial connective tissue in the K+ group can return to regularity and improve at the P₃ dose. The administration of ZAM showed a significant difference in cervical tissue after benzopyrene injection. <b>Conclusion:</b> Andaliman (<i>Zanthoxylum acanthopodium</i>) extract increases PI3K expression through suppression of Wnt expression. It can be developed therapy molecularly to prevent cell growth into cancer.

Correlation of Myc Expression with Wee1 Expression by Zanthoxylum acanthopodium in Cervical Carcinoma Histology.

<b>Background and Objective:</b> Natural herbs and molecular therapy can be used to treat cervical cancer. The Myc and Wee1 control tumour cell fate and microenvironmental changes like angiogenesis activation and host immune response suppression. The study aims to know about the correlation of Myc and Wee1 expressions as a molecular therapy given by <i>Zanthoxylum acanthopodium</i>. <b>Materials and Methods:</b> There are five rat groups: Group K is the untreated group, Group K⁺ is the rats injected with benzopyrene, Group P₁ is the administration of <i>Zanthoxylum acanthopodium</i> 100 mg kg¹ b.wt., Group P₂ is the administration of <i>Zanthoxylum acanthopodium</i> 200 mg kg¹ b.wt. and Group P₃ is the administration of <i>Zanthoxylum acanthopodium</i> 400 mg kg¹ b.wt. The rats are dissected 30 days after receiving <i>Zanthoxylum acanthopodium</i>. To stain the cervical tissues, immunohistochemistry is performed. <b>Results:</b> <i>Zanthoxylum acanthopodium</i> administration caused epithelial thickening and decreased Myc expression in previously uncontrolled carcinomas from untreated malignancies, which now slowed and stopped growing into the normal epithelium. Wee1 expression revealed that this herb could repair tissue by drastically reducing Wee1 expression at a dose of 100-400 mg kg¹ b.wt. Similarly, at the highest dose, cervical carcinoma stops growing and the nucleus begins to form normally (p<0.01). <b>Conclusion:</b> The higher Myc expression on andaliman administration in cervical carcinoma decreases Wee1 expression in cervical carcinoma so these two proteins have a strong and significant correlation. <i>Zanthoxylum acanthopodium</i> can be administered at various dosages to lower the number of positive indexes of Myc and Wee1 expression in cervical carcinoma.

Analysis of Apoptotic Cells and Lung Inflammation after Given by Vitis gracilis.

<b>Background and Objective:</b> Herbs have the potential to be used in molecular therapy to treat inflammation and apoptosis. It has been demonstrated that the Indonesian native <i>Vitris gracilis</i> inhibits cytochrome c as a precursor to apoptosis. When body tissues are injured, infected with bacteria, exposed to toxins or exposed to heat that causes cell death, inflammation occurs. The current study aimed to look for the effect of <i>Vitis gracilis</i> extract in the lung histology of mice. <b>Materials and Methods:</b> Five treatment groups were used for the experiments, which were conducted inside the study. The T1 is the negative control, T2 is swimming mice with 0.2 mg kg¹ b.wt., of vitamin C, T3 is swimming mice with 50 mg kg¹ b.wt., of <i>Vitis gracilis</i>, T4 is swimming mice with 75 mg kg¹ b.wt., of <i>Vitis gracilis</i> and T5 is swimming mice with 100 mg kg¹ b.wt., of <i>Vitis gracilis</i>. The dislocation procedure was used to dissect mice, ketamine was administered before dissection and the lungs were removed for TUNEL assay examination. <b>Results:</b> There were no significant differences in inflammatory cells or index-positive apoptotic cells between the T1, T2 and T5 groups (p>0.05). The T3 group had the highest value, while the T1 group had the lowest. The highest dose of <i>Vitis gracilis</i> reduced lung cell inflammation while also improving histological structure, resulting in intact, nucleated and complete alveolar membranes with proportional endothelial cells. <b>Conclusion:</b> <i>Vitis gracilis</i> 100 mg kg¹ b.wt., can repair and reduce inflammation in lung tissue. As a result, the higher the dose of <i>Vitis gracilis</i>, the less cell apoptosis occurs in the lungs.

Diabetic Wound Healing in FGF Expression by Nano Herbal of Rhodomyrtus tomentosa L. and Zanthoxylum acanthopodium Fruits.

<b>Background and Objective:</b> Increased glucose levels in diabetes mellitus patients affect angiogenesis which triggers the duration of the wound to heal. <i>Rhodomyrtus tomentosa</i> leaves (haramonting) and <i>Zanthoxylum acanthopodium </i>fruits (andaliman) are an endemic plant with an antioxidant in Indonesia. This study was aimed to determine histology changes of diabetic wound healing in FGF expression by Nano herbal of haramonting and andaliman. <b>Materials and Methods:</b> This study consisted of 4 groups for each treatment, K₁: Negative control, K₂: MEBO, K₃: Nano herbal of andaliman and K₄: Nano herbal of haramonting. The treatments were observed on days 0, 4, 8, 12 and 16. Diabetic model rats with alloxan injection (120 mg kg¹. IP) and rats were declared diabetes mellitus when blood glucose levels reached <u>></u>200 mg dL¹. The tissue was prepared on paraffin blocks and given Immunohistochemistry staining for FGF analysis. <b>Results:</b> There was a significant difference between all groups (p<0.001) in FGF expression. The proliferation of fibroblasts and collagen was formed by administering nano herbal andaliman and haramonting in rat's skin. The proliferation of cells that occur in the injured skin layer indicates the compounds contained in the nano herbal haramonting and andaliman stimulate cell division and growth to form wound tissue. <b>Conclusion:</b> Nano herbal andaliman and haramonting can be developed into herbs that can be used to treat wounds in diabetes. Another molecular gene analysis is required to get higher yields to further study for diabetic wounds against these two herbs.

Histological changes in placental rat apoptosis via FasL and cytochrome c by the nano-herbal Zanthoxylum acanthopodium.

Zanthoxylum acanthopodium has several biological activities, such as antimicrobial, anti-inflammatory, and antioxidant properties. This strong antioxidant herb can be used as a drug for hypertension. FasL and cytochrome c expression play roles in the apoptotic pathway in the placenta. This study focused on the histological change in apoptosis via cytochrome c and Fas ligand expression by investigating whether Zanthoxylum acanthopodium (ZA) fruits affect apoptosis. The present study consisted of five treatments: Normal pregnant rats (C), Hypertension rats (C + ), hypertension rats + extra virgin olive oil (EVOO) (T1), Hypertension rats + ZA (T2), and hypertension rats + EVOO + ZA (T3). Hypertension was induced in rats by injecting 3 mlml of 6% NaCl. Nanoherbal of ZA (100 mg/kg BW) and EVOO (1 ml) were given on the 13th-19th days of pregnancy. Pregnant rats were dissected on the 20th day of pregnancy by cervical dislocation. ELISA assays were performed for the analysis of HSP-70 expression. Immunohistochemistry and TUNEL assays were used to analyse the histological changes in placental tissue. The results showed that cytochrome c and FasL protein exposure levels in the labyrinth, basal, and yolk sac zones were increased during hypertensive pregnancy (P < 0.0001) in rats. The administration of nanoherbal of ZA decreased the expression of cytochrome c and FasL. A significant difference was found in the combination of nanoherbal of ZA and EVOO.

Effect of Nano Herbal Andaliman (Zanthoxylum acanthopodium) Fruits in NOTCH1 and Hes1 Expressions to Human Placental Trophoblasts.

BACKGROUND AND OBJECTIVE: Andaliman fruit (Zanthoxylum acanthopodium) is a well-known spice antioxidant in Northern Sumatera (Indonesia). The cellular activity requires antioxidants in counteracting free radicals. The cellular proteins that play a role in development, proliferation, differentiation and embryonic processes in the human placenta are NOTCH1 and Hes1. The aim of this research was to analyze the expression of NOTCH1 and Hes1 genes after administering nano herbal andaliman to the trophoblast cells of the human placenta. MATERIALS AND METHODS: HTR8 trophoblast cells were divided into two groups, namely, control and treatment (nano herbal andaliman). RNA isolation, reverse transcription and RT-PCR (real-time polymerase chain reaction) were performed to analyze the NOTCH1 and Notch target gene (Hes1) expressions. The NOTCH1 and Hes1 gene expressions were quantified using the CT method (2-ΔΔCT) and normalized with Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expressions. RESULTS: Nanoherbal andaliman reduced the expression of NOTCH1 genes in the human placental trophoblast. However, it increased the expression of Hes1 when the incubation time was 16 hrs. CONCLUSION: Nanoherbal andaliman decreases the expression of genes that are crucial in hypoxia and free radicals in the placenta, namely, NOTCH1 and Hes1 increased after incubation for 16 hrs. Therefore, this herb needs to be evaluated further.

Effect of Andaliman (Zanthoxylum acanthopodium DC.) Methanol Extract on Rat's Kidney and Liver Histology Induced by Benzopyrene.

BACKGROUND AND OBJECTIVE: The andaliman fruits have anti-inflammatory, antioxidant activity and a strong inhibition in antitumor activity. The purpose of this study was to analyze the effect of extract andaliman on rat's kidney and liver histology induced by benzopyrene. MATERIALS AND METHODS: The rats model of cancer-induced benzopyrene. This research consists of 5 groups; K-: Control, K+: Cancer model rats, P1: A dose of 100 mg per b.wt. per day of andaliman, P2: A dose of 200 mg per b.wt. per day and P3: A dose of 400 mg kg-1 b.wt., per day for 30 days. On the 31st day, performed surgically on the subjects. RESULTS: There were significant differences in the value of narrowing of the renal tubules (p<0.001), kidneys cells necrosis (p<0.01), hydrophilic degeneration (p<0.001), parenchymatous degeneration (p<0.01) and necrosis (p<0.001) in the liver after given the extract andaliman. CONCLUSION: Andaliman methanol extract repairs the damage of the liver and kidney of rats induced by benzopyrene. Andaliman can be recommended as a drug to repair the necrosis in the liver and kidneys caused by cancer.

Effects of Nanoherbal Haramonting (Rhodomyrtus tomentosa) and Extra Virgin Olive Oil on Histology of Liver and Kidney of Preeclamptic Rats.

BACKGROUND AND OBJECTIVE: Preeclampsia (PE) is a life threatening disease prevailing in pregnant women in Indonesia. Impaired kidneys and livers function in PE has a high risk during pregnancy. This study aimed to determine the role of nanoherbal haramonting and Extra Virgin Olive Oil (EVOO) on kidney and liver safety in preeclampsia rats. Nanoherbal Rhodomyrtus tomentosa is a medicinal plant with antioxidant activity such as EVOO which acts as inhibitor of oxidation, reduces lipid peroxidation and increases the speed of epithelialization. MATERIALS AND METHODS: This study used pregnant rats (Ratus norvegicus) consisting of 5 treatments. Animal models of preeclampsia were made by injection of NaCl 6% 3 mL/day/kg b.wt., subcutaneously in pregnancy of 6-12 days and then given herbs in pregnancy at 13-19 days. The study group consisted of, C-: Normal pregnant rats, C+: PE rats, T1: PE rats were given EVOO, T2: PE rats were given nanoherbal haramonting and T3: PE rats were given EVOO and nanoherbal haramonting. Tissue histology was made by the paraffin method and staining Hematoxylin Eosin which was dissected on the 20th day of pregnancy. RESULTS: There was no significant difference in weight of the liver and kidneys after administration of EVOO and nanoherbal haramonting. The administration of this herb significantly decreased the narrowing of the kidney tubules and the glomerular diameters. Both of these herbs also repaired preeclamptic liver damage (p<0.05) in normal hepatocyte cells, parenchymal degeneration and necrosis. CONCLUSION: The EVOO and nanoherbal haramonting repaired preeclamptic liver and kidney damage.

Effect of Nanoherbal Andaliman (Zanthoxylum acanthopodium) and Extra Virgin Olive Oil Combination on Preeclamptic Rats Liver Histology.

BACKGROUND: Andaliman (Zanthoxylum acanthopodium) is a spice traditional Northen Sumatera, Indonesia and these fruits contain alkaloids, steroids and terpenoids. Extra Virgin Olive Oil (EVOO) contains antioxidants. Combination of this plant have activities to reduce preeclampsia. AIM: To know the safety of the combination of nano herbal andaliman and Extra virgin olive oil (EVOO) on preeclampsia patients' liver. METHODS: Pregnant rats were made to have preeclampsia with 3 ml of NaCl 6% injections. This research consists of 5 groups: K- (negative control): normal pregnant rats, K+: preeclampsia rats; P1: PE rats were given nano herbal andaliman 1 ml EVOO / day / 20 gBW from the 13th to the 19th day of pregnancy, P2: PE rats were given nano herbal andaliman 100 mg/day / 200 gBW from the 13th to the 19th day of pregnancy, P3: PE was given the combination of 1 ml EVOO/day / 200 gBW and andaliman nano herbal 100 mg/day/200 gBW on the 13th day of pregnancy to the 19th day given orally. Then on the 20th day of pregnancy, the subjects were dissected. RESULTS: There were significant differences (p < 0.05) on the value of SGOT, SGPT, and the average damage of the hepatocyte cells except parenchymatous degeneration after being given the nano herbal andaliman and EVOO. The compared mean of normal hepatocytes cell, hydropic degeneration and necrosis value between all groups were p < 0.05 and p < 0.01 compared to (K-). The non-significant difference was found in the mean of parenchymatous degeneration between the groups (p = 0.058). CONCLUSION: The combination of nano herbal andaliman (Zanthoxylum acanthopodium) and EVOO affected the level of necrosis in hepatocyte cells on preeclampsia rats.

The Effect of Haramounting Leaf Ethanol Extract (Rhodomyrtus tomentosa (Aiton) Hassk.) on the Number of Leukocyte Type and Histology of Mice Pulmo (Mus Musculus L.) Exposed to Electronic Cigarette.

BACKGROUND: Haramonting leaves have a large antioxidant activity as inhibitors oxidation and form non-free radical compounds that are not reactive and relatively stable. Electronic smoking is a choice who slowly want to quit conventional smoking. AIM: This research aimed to determine the effect of ethanol extract on hard mounting leaves on leukocyte counts and histology Pulmo on male mice exposed to an electronic cigarette. METHODS: The samples used in this study were healthy male mice aged 8-11 weeks and weight of 25-30 g as many as 25 individuals and ethanol extracts of hard mounting leaves with a dose of 100 mg/kgBW, 200 mg/kgBW and 300 mg/kgBW. Blood tests were carried out at the Health Laboratory of North Sumatra Office, and the preparation of Pulmo histology was carried out at the Pathology and Anatomy Laboratory of Adam Malik General Hospital Medan. RESULT: There were significant differences between the control group and all of the treatment groups in the number of lymphocytes, neutrophils, basophils and eosinophils. There was a tendency that electronic cigarette smoke could cause damage to pulmo tissue. CONCLUSION: The results of this study indicate that the ethanol extract of hard mounting leaves could help the immune system in mice exposed to an electronic cigarette.

Interleukin-10 levels in Epstein-Barr virus-associated nasopharyngeal carcinoma.

The aim of this study was to determine the levels of interleukin-10 in patients with nasopharyngeal carcinoma. Both biopsies and sera were obtained from patients with nasopharyngeal carcinoma as well as Epstein-Barr virus-seronegative patients as a control. Nasopharyngeal carcinoma patients were classified using the World Health Organization pathological assessment and clinical staging of nasopharyngeal carcinoma. The numbers of interleukin-10 positive cells and the levels of serum interleukin-10 were assessed by immunohistochemical methods and enzyme-linked immunosorbent assay, respectively. The levels of serum interleukin-10 were determined by enzyme-linked immunosorbent assay. The results showed that the number of interleukin-10 positive cells and serum interleukin-10 levels were significantly increased in patients with nasopharyngeal carcinoma-World Health Organization type III and with clinical late stage (p<0.05), suggesting that interleukin-10 may have a crucial role in the progression of nasopharyngeal carcinoma.