Etlingera hemisphaerica Alters One-Dimensional Profile of Serum Proteins Due to Mercury Chloride in Rats (Rattus norvegicus).

<b>Background and Objective:</b> Several previous studies have shown that leaf ethanolic extract of <i>Etlingera hemisphaerica</i> (LE3H) has the potential to reduce the toxicity and teratogenicity effects of mercury. This study aimed to describe the effects of LE3H on the protein profile of <i>Rattus norvegicus</i> serum due to treatment with HgCl₂. <b>Materials and Methods:</b> Four groups of male rats, K1 was injected intraperitoneally (IP) HgCl₂ (5 mg kg¹ b.wt.), K2 was injected IP HgCl₂ (5 mg kg¹ b.wt.) and after 24 hrs it was gavage LE3H (0.27 mg g¹ b.wt.) every day for seven days, K3 was injected IP HgCl₂ (5 mg kg¹ b.wt.), after 24 hrs was gavage LE3H (0.55 mg g¹ b.wt.) every day for seven days. The K0 as control, received double-distilled water. On the ninth day, the experimental animals were killed via CD and blood was drawn from the heart to obtain serum. Serum samples were measured for protein content using the Lowry and serum was separated using the One-Dimensional Sodium dodecyl Sulfate-Polyacrylamide gel Electrophoresis (1D SDS-PAGE) technique. <b>Results:</b> The electropherograms showed four bands, 264.77, 219.53, 98.57 and 37.29 kDa, whose intensity significantly increased due to HgCl₂ treatment and then decreased to close to the control condition with LE3H administration. The results also revealed four bands, 31.95, 28, 06, 26, 29 and 15.09 kDa, whose intensity decreased significantly due to HgCl₂ treatment and then increased to close to the control condition by LE3H administration. <b>Conclusion:</b> The LE3H change profile of the eight blood serum protein bands due to HgCl₂ approximates the control condition in <i>R. norvegicus</i>.

Potential of Leave and Fruit Ethanolic Extract of Etlingera hemisphaerica as Antihyperuricemic in Mice (Mus musculus).

<b>Background and Objective:</b> Hyperuricemia is a disease triggered by disorders of uric acid metabolism. Therefore, this study evaluated the potential of leaves ethanolic extract of <i>Etlingera hemisphaerica</i> (LE3H) and fruits ethanolic extract <i>E. hemisphaerica</i> (FE3H) to restore hyperuricemia in mice. <b>Materials and Methods:</b> Six groups (A0, A1, A2, A3, A4 and A5) each consisted of four male mice. Hyperuricemia in mice was induced by giving 0.3 mL of fresh chicken liver juice (FCLJ) for seven days (A1, A2, A3, A4 and A5). The condition of hyperuricemia in A1 is not neutralized. Meanwhile, hyperuricemia conditions in A2, A3, A4 and A5 were neutralized for seven days by giving 0.01 mg g¹ body weight (BW) allopurinol, 0.13, 0.26 and 0.36 mg g¹ BW LE3H. The control group (A0) only received double-distilled water in the same way. Blood uric acid levels were measured with the GCU Meter Device before and after the induction of hyperuricemia and after efforts to neutralize the hyperuricemia condition. Six groups (B0, B1, B2, B3, B4, B5) each consisting of four male mice were also provided to test the potential of FE3H. The recovery potential FE3H against hyperuricemia was tested separately in the same way as was done for LE3H. <b>Results:</b> Giving FCLJ significantly increased (140.00-187.00%) uric acid compared to the control, so hyperuricemia was achieved. Doses of 0.13, 0.26 and 0.36 mg g¹ BW LE3H significantly recovered hyperuricemia as much as 54.09, 56.14 and 60.88%, respectively. Meanwhile, doses of 0.13, 0.26 and 0.36 mg g¹ FE3H significantly recovered hyperuricemia as much as 60.37, 62.24 and 65.572%, respectively. The LE3H and FE3H at the same dose showed that FE3H had a higher potential to restore hyperuricemia than LE3H. <b>Conclusion:</b> Leave and fruit ethanolic extract of <i>E. hemisphaerica</i> can potentially restore hyperuricemia in mice.

Leaf ethanolic extract of Etlingera hemesphaerica Blume mitigates defects in fetal anatomy and endochondral ossification due to mercuric chloride during the post-implantation period in Mus musculus.

This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) in reducing defects in fetal anatomy and endochondral ossification in mice induced by HgCl2 during the post-implantation period. Pregnant mice were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first group was administered LE3H (E1), the second one HgCl2 (E2), the third one HgCl2+LE3H (E3), and the fourth was control (E0), administered double-distilled water only. HgCl2 (5 mg/kg bw) was administrated by injection intraperitoneally on gestation day (GD)9 and LE3H (0.39 mg/g bw) was administered by gavage on GD10. The treated and control animals were killed by cervical dislocation on GD18, dissected, and the morphologically normal living fetuses (MNLF) were collected. The MNLF of E0, E1, E2, and E3 from 5 dams were fixed with Bouin solution, and observed using the free hand razor blade technique for soft tissue examination. The remaining MNLF were fixed with 96% ethanol, and then stained with Alizarin Red S and Alcian Blue for ossification examination. Index of length of ossified part (ILOP) of humerus, index of width of ossified part (IWOP) of humerus, ILOP of femur, and IWOP of femur were calculated. E2 had higher cases of anatomical defects (74,6%) than E3 (48.9%), E1 (15.0%), and E0 (0%). E2 had humerus IWOP of 0.82±0.03, which was significantly lower than that of E0 (0.89±0.04) and E1 (0.89±0.03), while that of E1 and E0 was not significantly different from each other. Meanwhile, IWOP in E3 (0.88±0.03) was significantly higher than that in E2, but not different from that in E1 and E0. Thus, LE3H mitigated defects in fetal anatomy and endochondral ossification induced by HgCl2 in mice.

Leaf ethanolic extract of Etlingera hemesphaerica Blume alters mercuric chloride teratogenicity during the post-implantation period in Mus musculus.

This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) against the teratogenic effects of mercuric chloride (HgCl2) in mice (Mus musculus). Pregnant M. musculus were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first, second, and third, and fourth (control) groups were administered with LE3H, HgCl2, HgCl2 + LE3H, and double-distilled water alone, respectively. HgCl2 (5 mg/kg bw) was administered by injection on gestation day (GD) 9, and LE3H (0.39 mg/g bw) was administered by gavage on GD 10. Treated and control animals were killed by cervical dislocation on GD 18, dissected, and the fetuses were collected for evaluation of maternal, embryonic, and fetal toxicity. Eight parameters were measured: (a) embryo resorption or resorbed embryo, (b) dead fetus, (c) living fetus, (d) morphologically normal living fetus, (e) malformed living fetus, (f) number of MLF, (g) length of MNLF, and (h) weight of MNLF. LE3H caused 4 (50.00%), whereas HgCl2 resulted in 7 (87.50%) parameters that were significantly different from those of the control, indicating that the teratogenicity of HgCl2 was significantly higher than that of LE3H. HgCl2 + LE3H showed two effects of LE3H on the teratogenicity of HgCl2: increased 2 (25.00%), and decreased 6 (75.00%). Thus, LE3H decreased the teratogenic effects of HgCl2 in M. musculus.

Preliminary studies on therapeutic effect of ethanolic extract of Tylophora villosa leaves against paracetamol-induced hepatotoxicity in mice.

This study intended to investigate the therapeutic effect of ethanolic extract of Tylophora villosa leaves (E2TL) against paracetamol (PC)-induced hepatotoxicity (PCIH) in mice (Mus musculus). PCIH were generated using daily 250 mg/kg body weight (bw) PC administration by gavage for seven days, and then daily 27.5; 55.0; 82.5; 110.0; or 220.0 mg/kg bw E2TL were treated by gavage for seven or fourteen days. Meanwhile, the controls were given solvent only in the same manner. Mortality, blood glucose, and condition (color, weight, volume) of the livers were observed on day 15 (D15). Serum glutamate pyruvate transaminase (SGPT) and serum glutamate oxaloacetate transaminase (SG0T) were examined on D15, D22, and D30, and then malondialdehyde (MDA) was determined on D15. Results of this study revealed that on D15, the dosage of 110.0 mg/kg bw E2TL most effectively decreased MDA due to PCIH, from 6.78 ± 1.70 µmol/L to 3.45 ± 0.43 µmol/L, approaching the control condition (2.45 ± 0.05 µmol/L). PC administration was really toxic dosage and caused 13.3 % mortality. Blood glucose, weight, and volume of the liver decreased as the effect of PC administration, and then 220.0 mg/kg bw E2TL treatment could recover the condition as well as the controls. Color of the liver indicated a similar recovery by E2TL treatment. SGPT and SG0T increased significantly by PC administration, and this PCIH facts could be recovered gradually near the controls according to the dosages (55.0; 110.0; or 220.0 mg/kg bw) and duration (seven or fourteen days) of E2TL treatment. It could be concluded that E2TL showed therapeutic effect against PCIH in M. musculus.

Protective Effect of Leaf Ethanolic Extract Etlingera hemisphaerica Blume Against Mercuric Chloride Toxicity in Blood of Mice.

This research was intended to investigate the protective effect of leaf ethanolic extract Etlingera hemisphaerica Blume (LE3H) against mercuric chloride (HgCl2) toxicity in blood of mice (Mus musculus). The experimental animals, 95 male M. musculus, received drink and food ad libitum. Three materials were tested: LE3H (0.13, 0.26, 0.39 mg/g body weight [bw]) was administered by gavage; HgCl2 (5 mg/kg bw) was administrated by gavage or intraperitoneal injection; and Imunos (the nutritional supplement to stimulate the immune system; 0.2 mg/g bw), as a positive control for LE3H treatment, was given by gavage. Blood samples were taken from the tails for determining number of blood cells. The animals were killed by cervical dislocation (CD), and then blood samples were collected from the hearts for protein electrophoresis. Results revealed the same number of leukocytes with LE3H (0.39 mg/g bw) treatment as with the Imunos treatment. HgCl2 administration increased leukocytes and decreased erythrocytes; HgCl2 administration followed by LE3H (0.39 mg/g bw) treatment protected the amount of blood cells as well as the control. HgCl2 administration showed a new 125 kDa protein and caused overexpression of 48 kDa protein; this protein profile could be protected by LE3H (0.39 mg/g bw) treatment as in the control condition. We conclude that LE3H provides a protective effect against HgCl2 toxicity in blood of M. musculus.

A teratoproteomics analysis: heat shock protein 70 is upregulated in mouse forelimb bud by methoxyacetic acid treatment.

BACKGROUND: Methoxyacetic acid (MAA) causes fetal limb abnormalities when the substance is administrated on gestation day (GD) 11 in mice. Limb abnormalities are caused mainly by extensive cell death in the mesoderm of the limb plate. This investigation focused on identifying a protein that is linked with mouse limb teratogenicity. METHODS: A single dose of MAA at 10 mmol/kg body weight was administered by gavage on GD 11; controls were administered vehicle only. Dams were killed by cervical dislocation 4 hr after treatment and forelimb buds were isolated from both the control and treated embryos. Proteins in forelimb buds GD 11 + 4 hr were precipitated out using 40-60% ammonium sulfate and were then analyzed by 2D SDS-PAGE. Excised protein spots were identified by mass spectrometry and amino acid internal sequence analysis. Identified protein was further confirmed by Western blotting. RESULTS: Two-dimensional gel analysis indicated that 1 protein spot of 81.7 kDa/pI 7.3 was overexpressed, and the protein matched heat shock protein 70 (HSP70; accession no. P08109, SwissProt). CONCLUSIONS: The results suggest that MAA, when administered to pregnant mice, upregulates HSP70 in the forelimb buds.

The laminin binding protein p40 is involved in inducing limb abnormality of mouse fetuses as the effects of methoxyacetic acid treatment.

This study is intended to characterize a protein that is linked with mouse limb teratogenicity as the effects of methoxyacetic acid (MAA) treatment. A single dose of MAA (10 mmol/kg body weight) was given by gavage on gestation day (GD) 11, whereas the control group were administered vehicle only. The pregnant mice were killed at 4 h after MAA treatment, and forelimb buds were isolated from both the control and treated group embryos. Proteins from forelimb buds GD 11 + 4 h, which were precipitated out using 40-60% ammonium sulfate, then were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2-D SDS-PAGE) technique. The 2-D gels reveal one protein with 41.6 kDa and pI 6.4, which expression was downregulated after MAA treatment. Tentative protein identification via peptide mass database search and definitive protein identification via a primary sequence database search indicate that the protein matches exactly to 34/67 kDa laminin binding protein (LBP; P14206, SwissProt), which is encoded by p40 gene (MGI:105381). The identity was further verified by Western blotting with an antibody against the 67 kDa LBP. The results suggest that MAA treatment to pregnant mice downregulates the LBP-p40 in the forelimb buds.