Research Article - Journal of Drug and Alcohol Research ( 2023) Volume 12, Issue 3
Characterization and Evaluation of Trans-Dermal Patches of Acalypha indica in Nanobubbles for Rheumatoid Arthritis
Kishore Babu1*, Ragi Mercy Rani2 and Rajiv Rajpal kukkar32Department of Pharmacy, St.Xavier Institute of Pharmacy, India
3Department of Pharmacy, Raffles University, India
Kishore Babu, Department of Pharmacy, KPJ Healthcare University, Malaysia, Email: babukishore@yahoo.co.in
Received: 29-Mar-2023, Manuscript No. JDAR-23- 99429; Editor assigned: 31-Mar-2023, Pre QC No. JDAR-23- 99429 (PQ); Reviewed: 14-Apr-2023, QC No. JDAR-23- 99429; Revised: 19-Apr-2023, Manuscript No. JDAR-23- 99429 (R); Published: 26-Apr-2023, DOI: 10.4303/JDAR/236235
Abstract
Herbal medicine is the oldest medicine that has been following from olden days to still now. It plays a major role in treating of different types of illness mostly in relieving of pain and from the conventional strategies are being used from the traditional medicine like ayurveda, unani and siddha. This is a custom therapy that has become fundamental now a days and has been the preferable one in the treating of wellspring of clinical treatment. Acalypha indica is the weed plant that has a wide range of medicinal qualities for the human well-being and it can be regularly seen in areas of India Pakistan, and Thailand. Chemical constituents are found in different parts of the plant and they are utilized for treating of different types of illness like eye contaminations, respiratory sickness, skin issues etc. Soxhlet apparatus is used for getting accurate and more reliable results. Paw induced oedema model is used to determine the analgesic activity. The 4 different types of extracts are used to determine analgesic activity by grouping the animals into 12 groups with different doses of 200 mg/kg and 400 mg/ kg are carried out in these groups Carrageenan control, AIEAF (200 mg/ kg and 400 mg/kg), AIAF (200 mg/kg and 400 mg/kg), AIEF (200 mg/kg and 400 mg/kg), AIMF (200 mg/kg and 400 mg/kg), AIWF (200 mg/kg), Diclofenac sodium (10 mg/kg) as standard reference and with different doses and compared the results to determine the analgesic effect. Results were plotted by using one-way Annova.
Keywords
Acalypha indica; Illness; Respiratory sickness
Introduction
Herbal medicine is the oldest trend in treating of various diseases and now it has been becoming as a latest trend because of the adverse effects caused by the long term use of Drugs. There are so many Indian medicinal plants available with different types of therapeutic properties. These plants posses’ different types of healing properties because of the different types of phytochemicals present in the plants. Many of these plants have already been identifies and discovered by different authors (Chopra et al., (1956), Nadkarni, (1976), Anonymous, (1985), Trease and Evans, (1985), Chatterjee, (1991), Kirtikar and Basu, (1995), Asolkar et al., (1992), Rastgi and Mehrotra, (1993), Elisabetsky et al., (1995), Warrier et al., (1996), Chatterjee, (2000), Joshi, (2000)), but the potency of these plants is yet to be proven therapeutically. Many of these plants were picked because of their therapeutic use in infectious diseases like fever, bronchitis, ulcer, diarrhoea, dysentery and skin diseases.
Inflammation and pain these 2 are considered to be the main manifestations of different diseases. NSAIDs are most commonly used in these types of conditions and they are induced occasionally. But, there is a severe adverse reaction caused by this type of drugs such as GIT Disturbances, Renal damage [1,2]. But now the analgesics are available with fewer side effects, the use of transdermal patches will avoid the chances of occurring of severe adverse effects and also helps in long term use of the medications.
Description
Plant
Acalypha indica is having different names like Acalypha indica (English: Indian Acalypha, Indian Mercury, Indian Copperleaf, Indian Nettle, Three-seeded Mercury) is an herbaceous annual that has catkin-like inflorescences with cup-shaped involucres surrounding the minute flowers. It is mainly known for its root being attractive to domestic cats, and for its various medicinal uses (Figure 1).
Figure 1: Acalypha indica plant
The plant is rich in cyanogenic glycoside called Acalyphin (a 3-cyanopyridone derivative) and also a flavonoid, some of the flavonoids are like clitorin, mauritianin, kaempferol glycosides [3]. The arial parts contain a cyanogenic glycoside called acalyphin (a 3-cyanopyridone derivative) as well as flavonoids, and biorobin, tanins, succinimide and flindersin (a pyranoquinolinone alkaloid has also been present). The chemicals that attract cats are the iridoid compounds isodihydronepetalactone and isoiridomyrmecin [4,5].
Methods
Collection of plant material and extraction
The plant was collected from the study area, washed thoroughly with tap water followed by shade dried then grounded in to coarse powder. The powdered plant material was extracted by maceration with methanol for 4 days with intermittent shaking. The extract was filtered through Whatman filter paper No. 1. The process was repeated three times to assure exhaustive extraction. The filtrates were concentrated in a rotatory evaporator (Buchi rotavapor, R-215) at 45°C to get the methanol extracts and kept at 4°C in refrigerator.
Preparation of extracts and fractionation of Acalypha indica
Solvent fractionation was used as a preliminary separation to simplify complex extracts with promising activity by fractionating chemical compounds into broad groups based on their solubility. The crude extracts were partially purified by solvent-solvent extraction method [6,7].
The methanol extract was suspended in water and portioned successively with ethyl acetate, acetone, ethanol, methanol and water. Ethyl acetate, acetone, ethanol and methanol in glass separating funnel, to give ethyl acetate, acetone, ethanol, methanol and residual water fractions respectively. Concentrated fractions were dried [8,9]. The extracts obtained were then subjected to various qualitative tests for the identification of various plant constituents and anti-inflammatory by Rat Paw Odema Method.
Evaluation of in vivo anti-inflammatory activity and grouping of animals
Carrageenan-induced paw odema model Paw odema was induced by injecting 0.1 ml of 1% w/v Carrageenan suspended in 1% CMC into sub-plantar tissues of the left hind paw of each rat. Rats were divided into 12 groups; each group consisting of 6 animals.
GROUP I: Carrageenan control
GROUP II: AIEAF (200 mg/kg)
GROUP III: AIEAF (400 mg/kg)
GROUP IV: AIAF (200 mg/kg)
GROUP V: AIAF (400 mg/kg)
GROUP VI: AIEF (200 mg/kg)
GROUP VII: AIEF (400 mg/kg)
GROUP VIII: AIMF (200 mg/kg)
GROUP IX: AIMF (400 mg/kg)
GROUP X: AIWF (200 mg/kg)
GROUP XI: AIWF (400 mg/kg)
GROUP XII: Diclofenac sodium (10 mg/kg) as standard reference.
The paw thickness was measured before injecting the Carrageenan and after 0 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes using Vernier caliper. The anti-inflammatory activity was calculated as percentage inhibition of odema in the animals treated with extract under test in comparison to the Carrageenan control group.
The percentage (%) inhibition of odema is calculated using the formula
where Tt is the thickness of paw of rats given test extract at corresponding time and To is the paw thickness of rats of control group at the same time.
Data analysis
The data is expressed as Mean ± Standard Deviation (SD). Results were analyzed using one-way ANOVA followed by Dunnet’s test. Differences were considered as statistically significant at P<0.05, when compared with control [10].
Results and Discussion
Carrageenan produced significant odema in the left hind paw of the vehicle treated group and the paw odema was significantly reduced in the standard drug, diclofenac sodium (10 mg/kg) treated group as a standard at all hours when compared to control group (Tables 1-3).
Table 1: Anti-inflammatory activity of extract of different solvents of Acalypha indica on Carrageenan induced paw odema method
| GROUP | RATS | Paw thickness (mm) after treatment of Carrageenan | ||||
|---|---|---|---|---|---|---|
| 0 h | 1 h | 2 h | 3 h | 4 h | ||
| GROUP-I | R1 | 1.4 | 3.4 | 4.4 | 6.4 | 4.8 |
| R2 | 1.8 | 3.8 | 4.9 | 6.6 | 4.9 | |
| R3 | 1.2 | 3 | 4.5 | 6.9 | 4.7 | |
| R4 | 1.3 | 3.2 | 4.7 | 6.4 | 4.6 | |
| R5 | 1.4 | 3.3 | 4.9 | 6.5 | 4.4 | |
| R6 | 1.3 | 3.4 | 4.7 | 6.4 | 4.5 | |
| MEAN | 1.4 | 3.35 | 4.68 | 6.53 | 4.65 | |
| SEM | 0.09 | 0.11 | 0.08 | 0.08 | 0.08 | |
| GROUP-II | R1 | 1.2 | 3 | 4.2 | 5.7 | 3.9 |
| R2 | 1.5 | 2.8 | 4.4 | 5.8 | 4.1 | |
| R3 | 1.2 | 3.1 | 4.2 | 5.9 | 4.2 | |
| R4 | 1.5 | 3.2 | 4 | 5.6 | 4 | |
| R5 | 1.3 | 3 | 4.2 | 5.8 | 3.9 | |
| R6 | 1.2 | 2.9 | 4.1 | 5.7 | 3.8 | |
| MEAN | 1.32 | 3 | 4.18 | 5.75 | 3.98 | |
| SEM | 0.06 | 0.06 | 0.05 | 0.04 | 0.06 | |
| GROUP-III | R1 | 1.4 | 3.1 | 4.4 | 5.8 | 3.8 |
| R2 | 1.6 | 2.9 | 4.1 | 5.9 | 3.5 | |
| R3 | 1.4 | 2.9 | 3.9 | 5.7 | 3.9 | |
| R4 | 1.8 | 3.2 | 4.4 | 6.1 | 3.4 | |
| R5 | 0.9 | 3.3 | 3.8 | 6.2 | 3.3 | |
| R6 | 1.2 | 3.1 | 3.7 | 5.9 | 3.8 | |
| MEAN | 1.38 | 3.08 | 4.05 | 5.93 | 3.62 | |
| SEM | 0.13 | 0.07 | 0.12 | 0.08 | 0.1 | |
| GROUP-IV | R1 | 1.2 | 2.9 | 4.3 | 5.9 | 4.1 |
| R2 | 1.5 | 3 | 4.3 | 5.8 | 3.9 | |
| R3 | 1.2 | 3.1 | 4 | 5.5 | 3.4 | |
| R4 | 1.5 | 3.1 | 4.4 | 4.9 | 3.8 | |
| R5 | 1.4 | 2.9 | 3.9 | 5.8 | 4.2 | |
| R6 | 1 | 2.9 | 3.7 | 5.6 | 3.8 | |
| MEAN | 1.3 | 2.98 | 4.1 | 5.58 | 3.87 | |
| SEM | 0.08 | 0.04 | 0.11 | 0.15 | 0.11 | |
| GROUP-V | R1 | 1.3 | 2.9 | 4.1 | 5.5 | 3.7 |
| R2 | 1.2 | 3 | 4.2 | 5.3 | 3.6 | |
| R3 | 1.7 | 3.5 | 4.3 | 5.1 | 3.9 | |
| R4 | 1.4 | 2.9 | 4 | 5.9 | 3.3 | |
| R5 | 1.1 | 2.8 | 3.9 | 5.8 | 3.5 | |
| R6 | 1.4 | 2.9 | 3.6 | 5.1 | 3.1 | |
| MEAN | 1.35 | 3 | 4.02 | 5.45 | 3.52 | |
| SEM | 0.08 | 0.1 | 0.1 | 0.14 | 0.12 | |
| GROUP-VI | R1 | 1.1 | 3 | 4.2 | 5.7 | 3.8 |
| R2 | 1.4 | 2.9 | 4.2 | 5.5 | 3.5 | |
| R3 | 1.2 | 2.8 | 3.9 | 5.2 | 3.7 | |
| R4 | 1.5 | 2.9 | 3.8 | 5.1 | 3.3 | |
| R5 | 1.4 | 2.9 | 3.5 | 4.9 | 3.2 | |
| R6 | 1.2 | 2.7 | 3.2 | 4.8 | 3.5 | |
| MEAN | 1.3 | 2.87 | 3.8 | 5.2 | 3.5 | |
| SEM | 0.06 | 0.04 | 0.16 | 0.14 | 0.09 | |
| GROUP-VII | R1 | 1.2 | 2.9 | 3.9 | 4.9 | 3.1 |
| R2 | 1.5 | 2.8 | 4.1 | 5.1 | 2.8 | |
| R3 | 1.5 | 3.1 | 3.9 | 4.8 | 3.4 | |
| R4 | 1.4 | 2.8 | 3.8 | 4.7 | 3.3 | |
| R5 | 1.1 | 2.7 | 3.1 | 4.9 | 3.3 | |
| R6 | 1.3 | 2.7 | 3.3 | 5.1 | 2.9 | |
| MEAN | 1.33 | 2.83 | 3.68 | 4.92 | 3.13 | |
| SEM | 0.07 | 0.06 | 0.16 | 0.07 | 0.1 | |
| GROUP-VIII | R1 | 1.1 | 3 | 4.2 | 5.2 | 3.2 |
| R2 | 1.4 | 2.9 | 4.2 | 4.9 | 3.5 | |
| R3 | 1.5 | 2.8 | 3.9 | 5 | 3.1 | |
| R4 | 1.5 | 2.9 | 3.8 | 4.8 | 2.9 | |
| R5 | 1.4 | 2.9 | 3.5 | 4.7 | 3.1 | |
| R6 | 1.2 | 2.7 | 3.2 | 4.8 | 2.5 | |
| MEAN | 1.35 | 2.87 | 3.8 | 4.9 | 3.05 | |
| SEM | 0.07 | 0.04 | 0.16 | 0.07 | 0.14 | |
| GROUP-IX | R1 | 1.2 | 2.9 | 3.5 | 3.8 | 2.7 |
| R2 | 1.1 | 2.8 | 3.3 | 4.2 | 2.8 | |
| R3 | 1.1 | 3.1 | 3.1 | 4.4 | 2.8 | |
| R4 | 1.2 | 2.8 | 3 | 4.1 | 2.5 | |
| R5 | 1.4 | 2.7 | 3.2 | 3.9 | 2.2 | |
| R6 | 1.7 | 2.7 | 3.1 | 3.9 | 2.8 | |
| MEAN | 1.28 | 2.83 | 3.2 | 4.05 | 2.63 | |
| SEM | 0.09 | 0.06 | 0.07 | 0.09 | 0.1 | |
| GROUP-X | R1 | 1.2 | 2.9 | 4.1 | 5.5 | 3.7 |
| R2 | 1.3 | 3 | 4.2 | 5.3 | 3.6 | |
| R3 | 1.8 | 3.5 | 4.3 | 5.1 | 3.9 | |
| R4 | 1.5 | 2.9 | 4 | 5.9 | 3.3 | |
| R5 | 1.1 | 2.8 | 3.9 | 5.8 | 3.5 | |
| R6 | 1.1 | 2.9 | 3.6 | 5.1 | 3.1 | |
| MEAN | 1.33 | 3 | 4.02 | 5.45 | 3.52 | |
| SEM | 0.11 | 0.1 | 0.1 | 0.14 | 0.12 | |
| GROUP-XI | R1 | 1.2 | 2.9 | 3.9 | 4.9 | 2.9 |
| R2 | 1.3 | 2.8 | 4.1 | 4.5 | 2.9 | |
| R3 | 1.5 | 3.2 | 3.5 | 4.4 | 3.3 | |
| R4 | 1.4 | 2.8 | 3.8 | 4.7 | 3.4 | |
| R5 | 1.1 | 2.7 | 3.1 | 4.9 | 3.5 | |
| R6 | 1.3 | 2.5 | 3.1 | 4.7 | 2.7 | |
| MEAN | 1.3 | 2.82 | 3.58 | 4.68 | 3.12 | |
| SEM | 0.06 | 0.09 | 0.17 | 0.08 | 0.13 | |
| GROUP-XII | R1 | 1.5 | 2.8 | 3.2 | 3.9 | 2.5 |
| R2 | 1.2 | 2.8 | 3.1 | 4 | 2.8 | |
| R3 | 0.9 | 2.9 | 3.1 | 4.1 | 2.4 | |
| R4 | 1.1 | 3 | 2.9 | 4.1 | 2.2 | |
| R5 | 1.2 | 2.9 | 3.2 | 4.2 | 2.8 | |
| R6 | 1.9 | 2.7 | 3.2 | 3.9 | 2.9 | |
| MEAN | 1.3 | 2.85 | 3.12 | 4.03 | 2.6 | |
| SEM | 0.14 | 0.04 | 0.05 | 0.05 | 0.11 | |
Table 2: Influence of selected extract of different solvents of Acalypha indica on Carrageenan induced paw odema method
| GROUP | Paw thickness (mm) after treatment of Carrageenan | ||||
|---|---|---|---|---|---|
| 0 h | 1 h | 2 h | 3 h | 4 h | |
| GROUP-I | 1.40 ± 0.09 | 3.35 ± 0.11 | 4.68 ± 0.08 | 6.53 ± 0.08 | 4.65 ± 0.08 |
| GROUP-II | 1.32 ± 0.06 | 3.00 ± 0.06* | 4.18 ± 0.05* | 5.75 ± 0.04*** | 3.98 ± 0.06** |
| GROUP-III | 1.38 ± 0.13 | 3.08 ± 0.07ns | 4.05 ± 0.012** | 5.93 ± 0.08*** | 3.62 ± 0.10*** |
| GROUP-IV | 1.30 ± 0.08 | 2.98 ± 0.04** | 4.10 ± 0.11** | 5.58 ± 0.15*** | 3.87 ± 0.11*** |
| GROUP-V | 1.35 ± 0.08 | 3.00 ± 0.10* | 4.02 ± 0.10** | 5.45 ± 0.14*** | 3.52 ± 0.12** |
| GROUP-VI | 1.30 ± 0.06 | 2.87 ± 0.04*** | 3.80 ± 0.16*** | 5.20 ± 0.14** | 3.50 ± 0.09*** |
| GROUP-VII | 1.33 ± 0.07 | 2.83 ± 0.06*** | 3.68 ± 0.16*** | 4.92 ± 0.07*** | 3.13 ± 0.10*** |
| GROUP-VIII | 1.35 ± 0.07 | 2.87 ± 0.04*** | 3.80 ± 0.16*** | 4.90 ± 0.07** | 3.05 ± 0.14*** |
| GROUP-IX | 1.28 ± 0.09 | 2.83 ± 0.06*** | 3.20 ± 0.07*** | 4.05 ± 0.09*** | 2.63 ± 0.10*** |
| GROUP-X | 1.33 ± 0.11 | 3.00 ± 0.10* | 4.02 ± 0.10** | 5.45 ± 0.14*** | 3.52 ± 0.12*** |
| GROUP-XI | 1.30 ± 0.06 | 2.82 ± 0.09*** | 3.58 ± 0.17*** | 4.68 ± 0.08*** | 3.12 ± 0.13** |
| GROUP-XII | 2.75 ± 0.14 | 2.85 ± 0.04*** | 3.12 ± 0.05*** | 4.03 ± 0.05*** | 2.60 ± 0.11*** |
| Values are the Mean ± S.E.M. of 6 rats/treatment; ***Significance P<0.001, **P<0.01,*P<0.05 when rest of groups compared wih Carrageenan control group (Group I). Data were analyzed by using One-way ANOVA followed by Dunnett’s test | |||||
Table 3: Percentage inhibition of paw odema by different solvent extracts of Acalypha indica on Carrageenan induced paw odema method
| GROUP | Paw thickness (mm) after treatment of Carrageenan | ||||
|---|---|---|---|---|---|
| 0 h | 1 h | 2 h | 3 h | 4 h | |
| GROUP-II | 6.33 | 11.67 | 11.95 | 13.62 | 16.74 |
| GROUP-III | 1.2 | 8.65 | 15.64 | 10.11 | 28.57 |
| GROUP-IV | 7.69 | 12.29 | 14.23 | 17.01 | 20.26 |
| GROUP-V | 3.7 | 11.67 | 16.6 | 19.88 | 32.23 |
| GROUP-VI | 7.69 | 16.86 | 23.25 | 25.64 | 32.86 |
| GROUP-VII | 5 | 18.24 | 27.15 | 32.88 | 48.4 |
| GROUP-VIII | 3.7 | 16.86 | 23.25 | 33.33 | 52.46 |
| GROUP-IX | 9.09 | 18.24 | 46.35 | 61.32 | 76.58 |
| GROUP-X | 5 | 11.67 | 16.6 | 19.88 | 32.23 |
| GROUP-XI | 7.69 | 18.93 | 30.7 | 39.5 | 49.2 |
| GROUP-XII | 7.69 | 17.54 | 50.27 | 61.98 | 78.85 |
The extract of Acalypha indica ethyl acetate fraction
It exhibited significant reduction in paw odema when compared to control group at 2 h and 4 h, at dose of 400 mg/kg b.w, p.o. The percentage of reduction was found to be 15.64 and 28.57 at 2 h and 4 h (Figures 2-7).
Figure 2: Effect of different solvent extracts of Acalypha indica on Carrageenan induced paw odema method at 0 h
Figure 3: Effect of different solvent extracts of Acalypha indica on Carrageenan induced paw odema method at 1 h
Figure 4: Effect of different solvent extracts of Acalypha indica on Carrageenan induced paw odema method at 2 h
Figure 5: Effect of different solvent extracts of Acalypha indica on Carrageenan induced paw odema method at 3 h
Figure 6: Effect of different solvent extracts of Acalypha indica on Carrageenan induced paw odema method at 4 h
Figure 7: Percentage inhibition of paw odema by different solvent extracts of Acalypha indica on Carrageenan induced paw odema method
The extract of Acalypha indica acetone fraction
It exhibited significant reduction in paw odema when compared to control group at 2 h, 3 h and 4 h, at dose of 400 mg/kg b.w, p.o. The percentage of reduction was found to be 16.66, 19.88 and 32.23 at 2 h, 3 h and 4 h.
The extract of Acalypha indica ethanol fraction
It exhibited significant reduction in paw odema when compared to control group at 2 h, 3 h and 4 h, at dose of 400 mg/kg b.w, p.o. The percentage of reduction was found to be 27.15, 32.88 and 48.40 at 2 h, 3 h and 4 h.
The extract of Acalypha indica methanol fraction
It exhibited significant reduction in paw odema when compared to control group at 1 h, 2 h, 3 h and 4 h, at dose of 400 mg/kg b.w, p.o. The percentage of reduction was found to be 18.24, 46.35, 61.32 and 76.58 at 1 h, 2 h, 3 h and 4 h.
The extract of Acalypha indica water fraction
It exhibited significant reduction in paw odema when compared to control group at 2 h, 3 h and 4 h, at dose of 400 mg/kg b.w, p.o. The percentage of reduction was found to be 30.70, 39.50 and 49.20 at 2 h, 3 h and 4 h.
Conclusion
From the above observations we may conclude that the methanol fraction of Acalypha indica more significantly reduced the paw odema from 1’ h onwards up to 4th h. the acetone, ethanol and water fraction of selected plant shows moderately significant reduction in the paw odema from 2’ h onwards up to 4th h. and the ethyl acetate fraction of selected plant shows less significant reduction of paw odema from 3rd h to 4th h.
We observed that the methanol soluble fraction of the selected plant shown significant anti-inflammatory activity when compared to the ethyl acetate, acetone, ethanol and water soluble fractions indicated that the polar constituents present in the methanol fractions may be contributing more towards the anti-inflammatory activity.
Acknowledgement
None.
Conflict of Interest
Authors have no conflict of interest to declare.
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