Aparna, P M
Propagation and bioactivity studies in Ellotti (Pterospermum rubiginosum b. Heyne ex wight & arn.) - Vellanikkara Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture 2024 - 173,xiip.
Ph.D
The study entitled “Propagation and bioactivity studies in Ellotti
(Pterospermum rubiginosum B. Heyne ex Wight & Arn.)” was conducted at the
Department of Plantation, Spices, Medicinal and Aromatic Crops, College of
Agriculture, KAU, Vellanikkara, and the College of Veterinary and Animal
Sciences, KVASU, Pookode, Wayanad from 2018 to 2023. The study aimed to
collect ethnobotanical information from tribal healers in Kerala, develop
vegetative propagation protocols, and investigate the bioactivity of Pterospermum
rubiginosum through antioxidant, antimicrobial, wound healing, and antihyperglycemic studies.
From the account of tribal healers, the bark of Pterospermum rubiginosum
is an effective remedy for the treatment of fractures. The tree's bark is effectively
used by the ethnomedical practitioners of different tribes in Kerala like
Kurichiyan, Kattunayakan, Muthuvan, Vetta Kuruman and Kanikkar. These
experienced healers assert that majority of patients respond well to the treatment,
approximately one in a hundred might exhibit allergic reactions or fail to respond
altogether. It may take a week to three months for a broken bone to mend. For a
fracture to heal, all factors like time taken for the patient to reach the healer,
severity of the fracture, patient's age, calcium status in the body and other health
conditions should be considered.
In the vegetative propagation study conducted during June-July and
September-October, maximum rooting was observed in the June-July months,
with the highest rooting percentage of 4.53 per cent. Among the growth
regulators, NAA 1000 ppm was found to be significantly superior in enchaining
the rooting in both hardwood and semi-hardwood cuttings of Pterospermum
rubiginosum at 120 days after planting.
Biochemical analysis of bark and leaves of Pterospermum rubiginosum
revealed the presence of phenols, flavonoids, tannins, steroids, saponins, sugars
and carbohydrates with the bark showing higher values compared to the leaves.
The gas chromatography-high resolution mass spectrometry (GC-HRMS) and
high resolution -liquid chromatography mass spectrometry (HR-LCMS) analysis
of ethanol extracts of bark and leaves revealed the presence of 22 major
compounds in the bark extract and 32 major compounds in the leaf extract. The
majority of the abundant compounds belong to flavonoid, phenol, alkaloid and
saponin class of compounds like 6''-caffeoylisoorientin, kaempferol 3-O-β-Dgalactoside, 3-O-cis-coumaroylmaslinic acid, epigallocatechin and patientoside
A.
The bark extract exhibited higher antioxidant activity in 2,2-diphenyl-2-
picrylhydrazyl hydrate (DPPH) radical scavenging assay, 2,2–azino–bis (3-
ethylbenzothiozoline-6-sulfonic acid) radical scavenging assay (ABTS), reducing
power assay, ferric reducing antioxidant power (FRAP) assay and total
antioxidant activity compared to the leaf extract, and similar activity to that of
leaf extract in nitric oxide scavenging assay.
The ethanol extract of both bark and leaves of Pterospermum rubiginosum
were ineffective against Staphylococcus aureus and Escherichia coli up to a
concentration of 1000 μg/mL in Kirby-Bauer agar disc diffusion method.
However, in modified resazurin microtiter plate assay, the ethanol extract of
Pterospermum rubiginosum bark exhibited minimum inhibitory concentration of
500 μg/mL against Staphylococcus aureus, while the leaf extract showed inhibition
against Escherichia coli at a minimum inhibitory concentration of 1750 μg/mL.
The ethanol extract of Pterospermum rubiginosum bark was used for
further in vitro studies as it showed superior biochemical, antioxidant and
antimicrobial properties compared to leaf extract. The per cent mean cell viability
of ethanol extract of Pterospermum rubiginosum bark in L929 mouse skin
fibroblast cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT) assay and resulted an effective concentration 50
(EC50) of 100.90 μg/mL. In scratch assay using L929 mouse skin fibroblast cells,
gap closure was at a faster rate in positive control, followed by test drug and
normal control. The in vitro test was followed by an in vivo wound healing study in
Wistar rats.
The per cent reduction in wound area on the 15th day of wound induction in
Wistar rats was similar in Cipladine standard (89.77 ± 2.39) and test extracts group
at 2.5 per cent (86.99 ± 1.17), 5.0 per cent (87.25 ± 1.91) and 10.0 per cent (85.99
± 1.01), which were significantly higher than normal control group (78.54 ± 2.93).
The presence of keratinocytes and incompletely formed hair follicles in
histopathological observations of Cipladine standard and extract 2.5 per cent group
suggested that the animals were in proliferative phase of wound healing whereas,
incomplete epithelialization in extracts 5.0 per cent, 10.0 per cent and normal
control led to the conclusion that the animals were in inflammatory phase of
wound healing.
In Wistar rats, further analysis using the oral glucose tolerance test (OGTT)
revealed that glucose values were higher one hour after glucose administration and
followed by a steady decrease after one hour. After three hours, the lowest glucose
values were observed in glibenclamide standard and extract at 50 mg/kg body
weight groups, whereas, normal control and higher concentration groups exhibited
significantly higher glucose values.
The study concludes that tribal healers effectively utilise Pterospermum
rubiginosum bark for healing bone fractures, wound and sprains. The ethanol
extract of Pterospermum rubiginosum bark and leaves exhibited antioxidant and
antimicrobial activity. In vivo wound healing studies suggested that a lower
concentration of extract at 2.5 per cent was effective in wound healing compared to
higher concentrations, possibly due to the presence of compounds hindering the
migration of proinflammatory cytokines to the wound site. Complete wound
healing could have been observed if the duration of study was increased to a few
more days. Another significant finding was the reduction in glucose level in Wistar
rats after administration of extract at 50 mg/kg body weight, which indicated that
Pterospermum rubiginosum has anti-hyperglycemic properties. Further in vivo
wound healing studies could be carried out by using bark extract at lower
concentrations or by isolation of compounds responsible for wound healing.
Plantation Spices Medicinal and Aromatic crops
Ellotti
Pterospermum rubiginosum b. Heyne ex wight & arn.
633.8 / APA/PR Ph.D
Propagation and bioactivity studies in Ellotti (Pterospermum rubiginosum b. Heyne ex wight & arn.) - Vellanikkara Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture 2024 - 173,xiip.
Ph.D
The study entitled “Propagation and bioactivity studies in Ellotti
(Pterospermum rubiginosum B. Heyne ex Wight & Arn.)” was conducted at the
Department of Plantation, Spices, Medicinal and Aromatic Crops, College of
Agriculture, KAU, Vellanikkara, and the College of Veterinary and Animal
Sciences, KVASU, Pookode, Wayanad from 2018 to 2023. The study aimed to
collect ethnobotanical information from tribal healers in Kerala, develop
vegetative propagation protocols, and investigate the bioactivity of Pterospermum
rubiginosum through antioxidant, antimicrobial, wound healing, and antihyperglycemic studies.
From the account of tribal healers, the bark of Pterospermum rubiginosum
is an effective remedy for the treatment of fractures. The tree's bark is effectively
used by the ethnomedical practitioners of different tribes in Kerala like
Kurichiyan, Kattunayakan, Muthuvan, Vetta Kuruman and Kanikkar. These
experienced healers assert that majority of patients respond well to the treatment,
approximately one in a hundred might exhibit allergic reactions or fail to respond
altogether. It may take a week to three months for a broken bone to mend. For a
fracture to heal, all factors like time taken for the patient to reach the healer,
severity of the fracture, patient's age, calcium status in the body and other health
conditions should be considered.
In the vegetative propagation study conducted during June-July and
September-October, maximum rooting was observed in the June-July months,
with the highest rooting percentage of 4.53 per cent. Among the growth
regulators, NAA 1000 ppm was found to be significantly superior in enchaining
the rooting in both hardwood and semi-hardwood cuttings of Pterospermum
rubiginosum at 120 days after planting.
Biochemical analysis of bark and leaves of Pterospermum rubiginosum
revealed the presence of phenols, flavonoids, tannins, steroids, saponins, sugars
and carbohydrates with the bark showing higher values compared to the leaves.
The gas chromatography-high resolution mass spectrometry (GC-HRMS) and
high resolution -liquid chromatography mass spectrometry (HR-LCMS) analysis
of ethanol extracts of bark and leaves revealed the presence of 22 major
compounds in the bark extract and 32 major compounds in the leaf extract. The
majority of the abundant compounds belong to flavonoid, phenol, alkaloid and
saponin class of compounds like 6''-caffeoylisoorientin, kaempferol 3-O-β-Dgalactoside, 3-O-cis-coumaroylmaslinic acid, epigallocatechin and patientoside
A.
The bark extract exhibited higher antioxidant activity in 2,2-diphenyl-2-
picrylhydrazyl hydrate (DPPH) radical scavenging assay, 2,2–azino–bis (3-
ethylbenzothiozoline-6-sulfonic acid) radical scavenging assay (ABTS), reducing
power assay, ferric reducing antioxidant power (FRAP) assay and total
antioxidant activity compared to the leaf extract, and similar activity to that of
leaf extract in nitric oxide scavenging assay.
The ethanol extract of both bark and leaves of Pterospermum rubiginosum
were ineffective against Staphylococcus aureus and Escherichia coli up to a
concentration of 1000 μg/mL in Kirby-Bauer agar disc diffusion method.
However, in modified resazurin microtiter plate assay, the ethanol extract of
Pterospermum rubiginosum bark exhibited minimum inhibitory concentration of
500 μg/mL against Staphylococcus aureus, while the leaf extract showed inhibition
against Escherichia coli at a minimum inhibitory concentration of 1750 μg/mL.
The ethanol extract of Pterospermum rubiginosum bark was used for
further in vitro studies as it showed superior biochemical, antioxidant and
antimicrobial properties compared to leaf extract. The per cent mean cell viability
of ethanol extract of Pterospermum rubiginosum bark in L929 mouse skin
fibroblast cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT) assay and resulted an effective concentration 50
(EC50) of 100.90 μg/mL. In scratch assay using L929 mouse skin fibroblast cells,
gap closure was at a faster rate in positive control, followed by test drug and
normal control. The in vitro test was followed by an in vivo wound healing study in
Wistar rats.
The per cent reduction in wound area on the 15th day of wound induction in
Wistar rats was similar in Cipladine standard (89.77 ± 2.39) and test extracts group
at 2.5 per cent (86.99 ± 1.17), 5.0 per cent (87.25 ± 1.91) and 10.0 per cent (85.99
± 1.01), which were significantly higher than normal control group (78.54 ± 2.93).
The presence of keratinocytes and incompletely formed hair follicles in
histopathological observations of Cipladine standard and extract 2.5 per cent group
suggested that the animals were in proliferative phase of wound healing whereas,
incomplete epithelialization in extracts 5.0 per cent, 10.0 per cent and normal
control led to the conclusion that the animals were in inflammatory phase of
wound healing.
In Wistar rats, further analysis using the oral glucose tolerance test (OGTT)
revealed that glucose values were higher one hour after glucose administration and
followed by a steady decrease after one hour. After three hours, the lowest glucose
values were observed in glibenclamide standard and extract at 50 mg/kg body
weight groups, whereas, normal control and higher concentration groups exhibited
significantly higher glucose values.
The study concludes that tribal healers effectively utilise Pterospermum
rubiginosum bark for healing bone fractures, wound and sprains. The ethanol
extract of Pterospermum rubiginosum bark and leaves exhibited antioxidant and
antimicrobial activity. In vivo wound healing studies suggested that a lower
concentration of extract at 2.5 per cent was effective in wound healing compared to
higher concentrations, possibly due to the presence of compounds hindering the
migration of proinflammatory cytokines to the wound site. Complete wound
healing could have been observed if the duration of study was increased to a few
more days. Another significant finding was the reduction in glucose level in Wistar
rats after administration of extract at 50 mg/kg body weight, which indicated that
Pterospermum rubiginosum has anti-hyperglycemic properties. Further in vivo
wound healing studies could be carried out by using bark extract at lower
concentrations or by isolation of compounds responsible for wound healing.
Plantation Spices Medicinal and Aromatic crops
Ellotti
Pterospermum rubiginosum b. Heyne ex wight & arn.
633.8 / APA/PR Ph.D