4.5 Antimicrobial activity Antimicr

4.5 Antimicrobial activity
Antimicrobial activity of ethanol and water extracts of Phyllanthus amarus were evaluated against the test organisms Salmonella typhi. Ethanolic, cold water extract and hot water extract of Phyllanthus amarus were employed for antimicrobial evaluation by agar cup diffusion method which were compared against standard antibiotics that were evaluated by disk diffusion method. The result demonstrates ethanolic extract to be most potent against the test bacteria with diameter of 8.0 mm as growth inhibition zone. This study establishes one of the traditional uses of Phyllanthus amarus against typhoid fever [26]. In another study, hexane, petroleum ether, chloroform, acetone and methanol extract of Phyllanthus leaves were tested for antibacterial activity against Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus mirabilis,
Streptococcus faecalis, Enterobacter species, Serratia marcescens, Staphylococcus aureus and Escherichia coli by agar well diffusion method. The results demonstrated methanol extract of Phyllanthus amarus for highest inhibitory activity against above bacterial species [27]. Similarly, in another study antimicrobial potential of Phyllanthus amarus were investigated using agar well diffusion method for activity against several drug resistant pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Klebsiella Species. The results revealed minimum inhibitory concentration (MIC) of the ethanolic plant extracts on Escherichia coli, Staphylococcus aureus, Pseudomona aeruginosa and Klebsiella Species were at 10 mg/ml, 50 mg/ml, 150 mg/ml and 100 mg/ml while the minimum bactericidal concentration were at 50 mg/ml, 100 mg/ml, 150 mg/ml and 150 mg/ml respectively [10]. Further studies on hexane, chloroform, ethyl acetate, acetone and methanol extract of stem bark extracts of Phyllanthus amarus demonstrated the antimicrobial activity for all these extracts with a diameter that ranges between 11 mm 24 mm against Streptococcus pyogenes, Staphylococcus aureus, Salmonella typhi, Escherichia coli, Candidas albican, Aspergillus flavus [8]. The antimicrobial activity of the methanolic extract of Phyllanthus amarus as studied by agar dilution method and disc diffusion showed significant concentration-dependent antibacterial activity specifically for gram-negative microbes.
It was also observed that antibacterial action was mainly due to the isolated phyllanthin [1]. These studies signify the antimicrobial potential of Phyllanthus amarus and need of isolation of some potential phytoconstituents from this species.

4.6 Antileptospiral Activity
Leptospirosis is globally important disease found mainly wherever human come in contact with the urine of infected animals or urine contaminated environment. Phyllanthus amarus have been investigated for the antileptospiral activity by micro dilution tests and tube dilution technique. The results revealed the inhibitory action of methanolic and aqueous extract of whole plant of Phyllanthus amarus against leptospira [28].

4.7 Anticonvulsant Activity
Epilepsy is a major neurological disorder characterized by the occurrence of recurrent seizures. The two widely proposed mechanisms involve alterations in the voltage-dependent ion channels such as reduction in inhibitory GABA-mediated drive or increase in excitatory glutamate mediated inputs. This chronic progressive CNS disorder affects a large population of the world. In search of herbal treatment, aqueous and ethanolic extract of Phyllanthus amarus were evaluated for anticonvulsant effect using pentylenetetrazole (PTZ) and maximal electroshock-induced seizures (MES) in swiss albino rats. The result showed ethanolic and aqueous extract of leaves and stem of Phyllanthus amarus significantly effective in abolishing hind limb extension induced by MES as well as PTZ induced seizures [29].

4.8 Antidiabetic Activity
Diabetes is a metabolic disorder of carbohydrate, fat and protein and is considered as the world’s largest endocrine disease [30]. The antidiabetic potential of Phyllanthus amarus investigated in an experiment model where fasted rats were made diabetic by single intraperitoneal injection of 120 mg/kg of alloxan monohydrates and then two doses of the aqueous and hydroalcoholic extract of Phyllanthus amarus administered orally which were then compared with the normal control group that received distilled water only. After 15 days treatment the result demonstrates aqueous and hydroalcoholic extract of Phyllanthus amarus decrease the blood glucose level significantly. Serum analysis of the treated experimental animals showed an increase in insulin and reduction in the malondialdehyde concentration, therefore demonstrated the potential antidiabetic property of aqueous and hydroalcoholic extract of Phyllanthus amarus [31]. In another study the methanolic extract of Phyllanthus amarus was found to inhibit lipid perxidation & scavenge hydroxyl and superoxide radicals [32]. Since free radicals are linked with diabetes, therefore quenching of free radical could be one mechanism of action [40]. However, there is a need of further experimental studies in order to isolate chemical constituents and their mechanism of action.

4.9 Anti-Inflammatory Activity
The anti-inflammatory potential of Phyllanthus amarus was evaluated using different models such as rat Kupffer cells, macrophages RAW264.7, human whole blood and in mice. Two different extracts of Phyllanthus amarus (hexane and ethanol/water extracts) and their anti-inflammatory effect was evaluated against the lipopolysaccharide stimulated above mentioned test cells. In addition, anti-inflammatory effect was evaluated in mice that were treated with galactosamine/lipopolysaccharide for inducing acute toxic hepatitis. The evaluation parameters were production of nitrite, prostaglandin E2 and cytokines that were measured by Griess assay, prostaglandin E2 by radioimmunoassay and latter by enzyme-linked immunosorbent assay. The other inflammatory markers such as endotoxin-induced nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) were determined by Western blot and activation of NF-κB and activator protein 1 (AP-1) by electrophoretic mobility shift assay (EMSA). The results revealed ethanol/water extracts and hexane extracts effective in inhibition of liopolysaccharide induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) in Kupffer cells and in macrophages RAW264.7. The extracts also attenuated the lipopolysaccharide induced secretion of tumor necrosis factor (TNF-α) in macrophages RAW264.7 as well as in human whole blood. Hexane and ethanol/water extracts of Phyllanthus amarus reduced expression of endotoxin-induced nitric oxide synthase iNOS and cyclooxygenase COX-2 and inhibited activation of nuclear factor NF-κB. Phyllanthus amarus also inhibited induction of interferon-γ (IFN-γ), interleukin (IL)-1β and interleukin (IL)-10 in human whole blood and reduced tumor necrosis factor (TNF-α) production in-vivo [33]. Further, experimental studies have been done to determine the chemical compounds responsible for the activity. In an attempt to test phytoconstituents and extracts of Phyllanthus amarus for anti-inflammatory effect, the purified lignans of Phyllanthus amarus and different extracts obtained from this plant were evaluated in carrageenan induced paw oedema and neutrophil influx model of inflammation. The result showed that hexane extract and the lignan-rich fraction, or lignans phyltetralin, nirtetralin and niranthin inhibited carrageenan-induced rat paw oedema, lower the increase of interleukin (IL)-1β tissue levels induced by carrageenan and inhibited neutrophil influx, bradykinin activating factor, platelet activating factor and endothelin-1induced paw oedema. These results show that the hexane extract, the lignan-rich fraction and the lignans niranthin, phyltetralin and nirtetralin exhibited marked anti-inflammatory properties [34]. Another interesting study where anti-inflammatory effect of soft drink prepared from the leaf extract of Phyllanthus amarus was evaluated for its anti-inflammatory effect and the result revealed anti-inflammatory activity of soft drink similar to reference compound Ibuprofen [35]. All these studies acknowledge Phyllanthus amarus as potent anti-inflammatory plant and lignins as potent phyto-compounds.

4.10 Antifertility Activity
Phyllanthus amarus possess antifertility activity. This activity was shown in the experimental study where alcoholic extract of Phyllanthus amarus brought changes in 3-beta and 17-beta hydroxyl steroid dehydrogenase (HSDs) levels, thereby effecting hormonal conversions in the female mice that confirmed by observation of no pregnancy in cohabited normal females and male mice [36].

4.11 Nephroprotective and cardioprotective activity Nephroprotective and cardioprotective effect of Phyllanthus amarus is evident from the study in which methanol extract of Phyllanthus amarus leaves caused a significant dose dependent decrease in the levels of total cholesterol, urea, total protein, uric acid, and prostatic, alkaline and acid phosphatases, aspartate transaminase (AST) and alanine transaminase (ALT) [37]. Since increase in these enzymes is related to hepatic and heart disorders therefore their reduction shows that the leaves of Phyllanthus amarus have hepato protective, nephroprotective and cardioprotective properties.

Kết quả (Việt) 1:

4,5 kháng khuẩn hoạt động Các hoạt động kháng khuẩn của ethanol và nước chiết xuất của Phyllanthus amarus được đánh giá chống lại bài kiểm tra sinh vật Salmonella typhi. Ethanolic, lạnh nước chiết xuất và bể nước chiết xuất Phyllanthus amarus được sử dụng để đánh giá kháng khuẩn bởi agar Cúp phương pháp phổ biến mà đã được so sánh với thuốc kháng sinh tiêu chuẩn đã được đánh giá bởi phương pháp phổ biến đĩa. Kết quả cho thấy ethanolic giải nén để mạnh nhất chống lại vi khuẩn thử nghiệm với đường kính 8,0 mm như tăng trưởng ức chế khu vực. Nghiên cứu này thiết lập một trong những sử dụng truyền thống của Phyllanthus amarus chống lại bệnh sốt thương hàn [26]. Trong nghiên cứu khác, hexane, dầu khí ête, cloroform, axeton và methanol trích xuất của Phyllanthus lá đã được thử nghiệm cho các hoạt động kháng khuẩn chống lại Pseudomonas aeruginosa, Klebsiella viêm phổi, Proteus mirabilis, Streptococcus faecalis, Enterobacter species, Serratia marcescens, Staphylococcus aureus and Escherichia coli by agar well diffusion method. The results demonstrated methanol extract of Phyllanthus amarus for highest inhibitory activity against above bacterial species [27]. Similarly, in another study antimicrobial potential of Phyllanthus amarus were investigated using agar well diffusion method for activity against several drug resistant pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Klebsiella Species. The results revealed minimum inhibitory concentration (MIC) of the ethanolic plant extracts on Escherichia coli, Staphylococcus aureus, Pseudomona aeruginosa and Klebsiella Species were at 10 mg/ml, 50 mg/ml, 150 mg/ml and 100 mg/ml while the minimum bactericidal concentration were at 50 mg/ml, 100 mg/ml, 150 mg/ml and 150 mg/ml respectively [10]. Further studies on hexane, chloroform, ethyl acetate, acetone and methanol extract of stem bark extracts of Phyllanthus amarus demonstrated the antimicrobial activity for all these extracts with a diameter that ranges between 11 mm 24 mm against Streptococcus pyogenes, Staphylococcus aureus, Salmonella typhi, Escherichia coli, Candidas albican, Aspergillus flavus [8]. The antimicrobial activity of the methanolic extract of Phyllanthus amarus as studied by agar dilution method and disc diffusion showed significant concentration-dependent antibacterial activity specifically for gram-negative microbes. It was also observed that antibacterial action was mainly due to the isolated phyllanthin [1]. These studies signify the antimicrobial potential of Phyllanthus amarus and need of isolation of some potential phytoconstituents from this species.

4.6 Antileptospiral Activity
Leptospirosis is globally important disease found mainly wherever human come in contact with the urine of infected animals or urine contaminated environment. Phyllanthus amarus have been investigated for the antileptospiral activity by micro dilution tests and tube dilution technique. The results revealed the inhibitory action of methanolic and aqueous extract of whole plant of Phyllanthus amarus against leptospira [28].

4.7 Anticonvulsant Activity
Epilepsy is a major neurological disorder characterized by the occurrence of recurrent seizures. The two widely proposed mechanisms involve alterations in the voltage-dependent ion channels such as reduction in inhibitory GABA-mediated drive or increase in excitatory glutamate mediated inputs. This chronic progressive CNS disorder affects a large population of the world. In search of herbal treatment, aqueous and ethanolic extract of Phyllanthus amarus were evaluated for anticonvulsant effect using pentylenetetrazole (PTZ) and maximal electroshock-induced seizures (MES) in swiss albino rats. The result showed ethanolic and aqueous extract of leaves and stem of Phyllanthus amarus significantly effective in abolishing hind limb extension induced by MES as well as PTZ induced seizures [29].

4.8 Antidiabetic Activity
Diabetes is a metabolic disorder of carbohydrate, fat and protein and is considered as the world’s largest endocrine disease [30]. The antidiabetic potential of Phyllanthus amarus investigated in an experiment model where fasted rats were made diabetic by single intraperitoneal injection of 120 mg/kg of alloxan monohydrates and then two doses of the aqueous and hydroalcoholic extract of Phyllanthus amarus administered orally which were then compared with the normal control group that received distilled water only. After 15 days treatment the result demonstrates aqueous and hydroalcoholic extract of Phyllanthus amarus decrease the blood glucose level significantly. Serum analysis of the treated experimental animals showed an increase in insulin and reduction in the malondialdehyde concentration, therefore demonstrated the potential antidiabetic property of aqueous and hydroalcoholic extract of Phyllanthus amarus [31]. In another study the methanolic extract of Phyllanthus amarus was found to inhibit lipid perxidation & scavenge hydroxyl and superoxide radicals [32]. Since free radicals are linked with diabetes, therefore quenching of free radical could be one mechanism of action [40]. However, there is a need of further experimental studies in order to isolate chemical constituents and their mechanism of action.

4.9 Anti-Inflammatory Activity
The anti-inflammatory potential of Phyllanthus amarus was evaluated using different models such as rat Kupffer cells, macrophages RAW264.7, human whole blood and in mice. Two different extracts of Phyllanthus amarus (hexane and ethanol/water extracts) and their anti-inflammatory effect was evaluated against the lipopolysaccharide stimulated above mentioned test cells. In addition, anti-inflammatory effect was evaluated in mice that were treated with galactosamine/lipopolysaccharide for inducing acute toxic hepatitis. The evaluation parameters were production of nitrite, prostaglandin E2 and cytokines that were measured by Griess assay, prostaglandin E2 by radioimmunoassay and latter by enzyme-linked immunosorbent assay. The other inflammatory markers such as endotoxin-induced nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) were determined by Western blot and activation of NF-κB and activator protein 1 (AP-1) by electrophoretic mobility shift assay (EMSA). The results revealed ethanol/water extracts and hexane extracts effective in inhibition of liopolysaccharide induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) in Kupffer cells and in macrophages RAW264.7. The extracts also attenuated the lipopolysaccharide induced secretion of tumor necrosis factor (TNF-α) in macrophages RAW264.7 as well as in human whole blood. Hexane and ethanol/water extracts of Phyllanthus amarus reduced expression of endotoxin-induced nitric oxide synthase iNOS and cyclooxygenase COX-2 and inhibited activation of nuclear factor NF-κB. Phyllanthus amarus also inhibited induction of interferon-γ (IFN-γ), interleukin (IL)-1β and interleukin (IL)-10 in human whole blood and reduced tumor necrosis factor (TNF-α) production in-vivo [33]. Further, experimental studies have been done to determine the chemical compounds responsible for the activity. In an attempt to test phytoconstituents and extracts of Phyllanthus amarus for anti-inflammatory effect, the purified lignans of Phyllanthus amarus and different extracts obtained from this plant were evaluated in carrageenan induced paw oedema and neutrophil influx model of inflammation. The result showed that hexane extract and the lignan-rich fraction, or lignans phyltetralin, nirtetralin and niranthin inhibited carrageenan-induced rat paw oedema, lower the increase of interleukin (IL)-1β tissue levels induced by carrageenan and inhibited neutrophil influx, bradykinin activating factor, platelet activating factor and endothelin-1induced paw oedema. These results show that the hexane extract, the lignan-rich fraction and the lignans niranthin, phyltetralin and nirtetralin exhibited marked anti-inflammatory properties [34]. Another interesting study where anti-inflammatory effect of soft drink prepared from the leaf extract of Phyllanthus amarus was evaluated for its anti-inflammatory effect and the result revealed anti-inflammatory activity of soft drink similar to reference compound Ibuprofen [35]. All these studies acknowledge Phyllanthus amarus as potent anti-inflammatory plant and lignins as potent phyto-compounds.

4.10 Antifertility Activity
Phyllanthus amarus possess antifertility activity. This activity was shown in the experimental study where alcoholic extract of Phyllanthus amarus brought changes in 3-beta and 17-beta hydroxyl steroid dehydrogenase (HSDs) levels, thereby effecting hormonal conversions in the female mice that confirmed by observation of no pregnancy in cohabited normal females and male mice [36].

4.11 Nephroprotective and cardioprotective activity Nephroprotective and cardioprotective effect of Phyllanthus amarus is evident from the study in which methanol extract of Phyllanthus amarus leaves caused a significant dose dependent decrease in the levels of total cholesterol, urea, total protein, uric acid, and prostatic, alkaline and acid phosphatases, aspartate transaminase (AST) and alanine transaminase (ALT) [37]. Since increase in these enzymes is related to hepatic and heart disorders therefore their reduction shows that the leaves of Phyllanthus amarus have hepato protective, nephroprotective and cardioprotective properties.