Evaluation of in vitro and in vivo therapeutic efficacy of Ribes alpestre Decne in Rheumatoid arthritis

Hasan, Alamgeer Umme Habiba

doi:10.1590/s2175-97902019000217832

Alamgeer Umme Habiba Hasan

Laboratory of Cardiovascular Research and Integrative Pharmacology, Department of Pharmacology, College of Pharmacy, University of Sargodha, Sargodha, Pakistan

http://orcid.org/0000-0003-4775-7337

*Correspondence: Laboratory of Cardiovascular Research and Integrative Pharmacology, Department of Pharmacology, College of Pharmacy, University of Sargodha, Sargodha, Pakistan. Email: alam_yuchi@yahoo.com

Percentage protection against protein denaturation Concentration (ug/mL)
Treatment Groups	50	100	200	400	800	1600	3200	6400
Aspirin	62.44 ± 0.558	64.66 ± 0.461	66.59 ± 0.391	69.33 ± 0.463	72.586 ± 0.451	74.663 ± 0.446	77.550 ± 0.463	80.586 ± 0.451
Aqueous ethanol extract	60.28 ± 2.356^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni post test	64.61 ± 1.84^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni post test	67.433 ± 2.225^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni post test	79.163 ± 0.80^* * p<0.05,	82.373 ± 1.061^* * p<0.05,	84.780 ± 0.400^* * p<0.05,	87.506 ± 1.926^* * p<0.05,	95.00 ± 0.577^* ***** p<0.001,
n-Butanol Fraction	35.623 ± 4.06^* ***** p<0.001,	40.126 ± 3.383^* ***** p<0.001,	45.260 ± 3.495^* ***** p<0.001,	55.993 ± 5.350^ p<0.01,	58.380 ± 2.839^* ***** p<0.001,	61.113 ± 2.627^ p<0.01,	65.510 ± 2.25^ p<0.01,	69.233 ± 2.126^* * p<0.05,
Aqueous Fraction	45.516 ± 0.803^* ***** p<0.001,	50.776 ± 4.364^* ***** p<0.001,	56.776 ± 4.364^* * p<0.05,	80.390 ± 1.807^* * p<0.05,	86.176 ± 1.958^ p<0.01,	85.246 ± 2.407^* * p<0.05,	87.846 ± 3.651^* * p<0.05,	92.840 ± 2.603^ p<0.01,

Percentage protection against protein denaturation Concentration (ug/mL)
Treatment Groups	50	100	200	400	800	1600	3200	6400
Diclofenac sodium	66.67 ± 5.24	67.36 ± 4.22	69.44 ± 1.386	77.07 ± 2.40	88.14 ± 1.88	182.64 ± 2.50	370.83 ± 2.08	717.36 ± 5.42
Aqueous ethanol extract	88.890 ± 4.862^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	127.08 ± 8.67^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	190.97 ± 17.27^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	458.33 ± 15.91^ p<0.01,	1122.92 ± 65.84^* ***** p<0.001,	1522.92 ± 136.40^* ***** p<0.001,	2022.91 ± 124.27^* ***** p<0.001,	3653.47 ± 200.92^* ***** p<0.001,
n-Butanol Fraction	37.50 ± 2.08^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	50.69 ± 1.386^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	65.97 ± 4.22^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	95.83 ± 4.34^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	213.19 ± 20.14^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	298.50 ± 1.50^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	703.22 ± 74.70^ p<0.01,	1484.03 ± 211.17^* ***** p<0.001,
Aqueous Fraction	140.28 ± 5.42^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	171.53 ± 4.86^ns ns non-significant as compared to Standard control done by Two way ANOVA followed by Bonferroni posttest.	400.00 ± 14.18^ p<0.01,	818.05 ± 19.59^* ***** p<0.001,	1625.00 ± 40.74^* ***** p<0.001,	1977.77 ± 25.52^* ***** p<0.001,	2204.17 ± 49.14^* ***** p<0.001,	3563.19 ± 128.48^* ***** p<0.001,

Percentage protection against membrane lysis Concentration (ug/mL)
Treatment Groups	50	100	200	400	800	1600	3200	6400
Diclofenac sodium	20.30 ± 0.144	21.79 ± 0.15	22.61 ± 0.17	24.56 ± 0.17	26.70 ± 0.14	31.36 ± 0.15	49.54 ± 0.10	70.50 ± 0.12
Aqueous ethanol extract	69.11 ± 3.34^* ***** p<0.001,	75.18 ± 0.51^* ***** p<0.001,	79.63 ± 2.56^* ***** p<0.001,	86.55 ± 0.32^* ***** p<0.001,	88.07 ± 0.44^* ***** p<0.001,	89.65 ± 1.06^* ***** p<0.001,	90.81 ± 0.38^* ***** p<0.001,	91.29 ± 0.42^* ***** p<0.001,
n-Butanol Fraction	44.45 ± 1.50^* ***** p<0.001,	48.90 ± 0.84^* ***** p<0.001,	52.86 ± 1.76^* ***** p<0.001,	57.30 ± 2.55^* ***** p<0.001,	59.21 ± 2.95^* ***** p<0.001,	62.97 ± 0.19^* ***** p<0.001,	64.77 ± 0.23^* ***** p<0.001,	65.73 ± 1.66^ns ns non-significant as compared to Standard control.
Aqueous Fraction	60.38 ± 2.51^* ***** p<0.001,	65.18 ± 6.25^* ***** p<0.001,	73.83 ± 0.73^* ***** p<0.001,	77.15 ± 0.36^* ***** p<0.001,	80.73 ± 0.09^* ***** p<0.001,	84.33 ± 0.27^* ***** p<0.001,	86.54 ± 0.09^* ***** p<0.001,	89.62 ± 0.07^* ***** p<0.001,

Treatment groups	2^nd day	4^th day	6^th day	8^th day	10^th day
Increase in paw volume (mL)
Arthritic control (3 mL/kg)	1.474±0.044	1.872±0.025	2.000±0.014	2.032±0.017	2.120±0.029
Piroxicam (10 mg/kg)	1.196±0.069^ p<0.01, (18.86%)	1.044±0.069^* ***** p<0.001, (44.23%)	0.852±0.068^* ***** p<0.001, (57.4%)	0.682±0.049^* ***** p<0.001, (66.44%)	0.596±0.042^* ***** p<0.001, (71.89%)
Aqueous ethanol extract (50 mg/kg)	1.248±0.011^* ***** p<0.001, (15.33%)	1.362±0.015^* ***** p<0.001, (27.24%)	1.300±0.008^* ***** p<0.001, (35%)	1.222±0.011^* ***** p<0.001, (39.96%)	1.106±0.007^* ***** p<0.001, (47.83%)
Aqueous ethanol extract (100 mg/kg)	1.100±0.011^* ***** p<0.001, (25.37%)	1.204±0.029^* ***** p<0.001, (35.68%)	1.150±0.029^* ***** p<0.001, (42.5%)	1.080±0.028^* ***** p<0.001, (46.85%)	1.000±0.028^* ***** p<0.001, (52.83%)
Aqueous ethanol extract (200 mg/kg)	0.998±0.066^* ***** p<0.001, (32.29%)	1.044±0.102^* ***** p<0.001, (44.23%)	0.786±0.046^* ***** p<0.001, (60.7%)	0.692±0.040^* ***** p<0.001, (65.94%)	0.562±0.010^* ***** p<0.001, (73.49%)
Butanol fraction (50 mg/kg)	1.306±0.043^ns ns non-significant (11.39%)	1.544±0.048^* ***** p<0.001, (17.52%)	1.450±0.064^* ***** p<0.001, (27.5%)	1.302±0.058^* ***** p<0.001, (35.93%)	1.220±0.051^* ***** p<0.001, (42.45%)
Butanol fraction (100 mg/kg)	1.224±0.045^* (16.96%)	1.468±0.067^* ***** p<0.001, (21.58%)	1.396±0.053^* ***** p<0.001, (30.2%)	1.326±0.042^* ***** p<0.001, (34.74%)	1.198±0.035^* ***** p<0.001, (43.49%)
Butanol fraction (200 mg/kg)	1.214±0.101^ p<0.01, (17.64%)	1.344±0.051^* ***** p<0.001, (28.21%)	1.084±0.089^* ***** p<0.001, (45.8%)	0.872±0.042^* ***** p<0.001, (57.09%)	0.712±0.039^* ***** p<0.001, (66.42%)
Aqueous Fraction (50 mg/kg)	1.254±0.062^ns ns non-significant (14.93%)	1.426±0.100^* ***** p<0.001, (23.82%)	1.350±0.078^* ***** p<0.001, (32.5%)	1.288±0.069^* ***** p<0.001, (36.61%)	1.208±0.086^* ***** p<0.001, (43.02%)
Aqueous Fraction (100 mg/kg)	1.170±0.065^* * p<0.05, (20.62%)	1.382±0.104^* ***** p<0.001, (26.18%)	1.320±0.081^* ***** p<0.001, (34%)	1.250±0.106^* ***** p<0.001, (38.48%)	1.166±0.114^* ***** p<0.001, (45%)
Aqueous Fraction (200 mg/kg)	1.150±0.079^ p<0.01, (21.98%)	1.278±0.043^* ***** p<0.001, (31.73%)	0.870±0.043^* ***** p<0.001, (56.5%)	0.788±0.070^* ***** p<0.001, (61.22%)	0.660±0.062^* ***** p<0.001, (68.87%)

Treatment groups	2^nd day	4^th day	6^th day	8^th day	10^th day
Increase in paw diameter (mm)
Arthritic control (3 mL/kg)	6.436±0.098	8.454±0.023	10.578±0.024	11.11±0.026	12.772±0.128
Piroxicam (10 mg/kg)	4.800±0.198^* ***** p<0.001, (25.42%)	5.742±0.303^* ***** p<0.001, (32.08%)	5.282±0.180^* ***** p<0.001, (50.07%)	4.102±0.094^* ***** p<0.001, (60.08%)	3.744±0.133^* ***** p<0.001, (70.69%)
Aqueous ethanol extract (50 mg/kg)	5.874±0.097^* * p<0.05, (8.73%)	7.226±0.078^* ***** p<0.001, (14.52%)	7.252±0.097^* ***** p<0.001, (31.44%)	7.222±0.106^* ***** p<0.001, (35%)	7.080±0.108^* ***** p<0.001, (44.56%)
Aqueous ethanol extract (100 mg/kg)	5.228±0.110^* ***** p<0.001, (18.76%)	5.810±0.183^* ***** p<0.001, (31.27%)	5.734±0.188^* ***** p<0.001, (45.79%)	5.644±0.177^* ***** p<0.001, (49.20%)	5.606±0.156^* ***** p<0.001, (56.11%)
Aqueous ethanol extract (200 mg/kg)	4.812±0.134^* ***** p<0.001, (25.23%)	4.808±0.161^* ***** p<0.001, (43.13%)	4.242±0.075^* ***** p<0.001, (59.89%)	3.866±0.122^* ***** p<0.001, (65.21%)	3.528±0.033^* ***** p<0.001, (72.38%)
Butanol fraction (50 mg/kg)	6.122±0.114^ns ns non-significant (4.88%)	8.216±0.272^ns ns non-significant (2.82%)	8.05±0.290^* ***** p<0.001, (23.89%)	7.584±0.328^* ***** p<0.001, (31.74%)	7.442±0.334^* ***** p<0.001, (41.73%)
Butanol fraction (100 mg/kg)	5.70±0.154^ns ns non-significant (3.76%)	7.422±0.189^ p<0.01, (12.21%)	6.872±0.293^* ***** p<0.001, (35.03%)	6.144±0.353^* ***** p<0.001, (44.70%)	5.930±0.369^* ***** p<0.001, (53.57%)
Butanol fraction (200 mg/kg)	5.504±0.131^* * p<0.05, (14.48%)	6.422±0.102^* ***** p<0.001, (24.04%)	6.550±0.119^* ***** p<0.001, (22.52%)	6.104±0.154^* ***** p<0.001, (45.06%)	5.760±0.159^* ***** p<0.001, (54.90%)
Aqueous fraction (50 mg/kg)	6.086±0.139^ns ns non-significant (5.44%)	7.092±0.144* (16.11%)	6.924±0.083^* ***** p<0.001, (34.54%)	7.002±0.164^* ***** p<0.001, (36.98%)	6.178±1.513^* ***** p<0.001, (51.63%)
Aqueous fraction (100 mg/kg)	5.248±0.068^ns ns non-significant (18.46%)	6.410±0.154^* ***** p<0.001, (24.18%)	6.506±0.112^* ***** p<0.001, (38.49%)	5.994±0.064^* ***** p<0.001, (46.05%)	5.696±0.081^* ***** p<0.001, (55.40%)
Aqueous fraction (200 mg/kg)	4.940±0.094^ p<0.01, (23.24%)	5.570±0.155^* ***** p<0.001, (34.11%)	5.600±0.173^* ***** p<0.001, (47.06%)	4.860±0.098^* ***** p<0.001, (56.26%)	4.300±0.116^* ***** p<0.001, (66.33%)

[1] *Correspondence: Laboratory of Cardiovascular Research and Integrative Pharmacology, Department of Pharmacology, College of Pharmacy, University of Sargodha, Sargodha, Pakistan. Email: alam_yuchi@yahoo.com

Treatment groups	2^nd day	4^th day	6^th day	8^th day	10^th day
Increase in paw volume (mL)
Arthritic control (3 mL/kg)	1.474±0.044	1.872±0.025	2.000±0.014	2.032±0.017	2.120±0.029
Piroxicam (10 mg/kg)	1.196±0.069^ p<0.01, (18.86%)	1.044±0.069^* ***** p<0.001, (44.23%)	0.852±0.068^* ***** p<0.001, (57.4%)	0.682±0.049^* ***** p<0.001, (66.44%)	0.596±0.042^* ***** p<0.001, (71.89%)
Aqueous ethanol extract (50 mg/kg)	1.248±0.011^* ***** p<0.001, (15.33%)	1.362±0.015^* ***** p<0.001, (27.24%)	1.300±0.008^* ***** p<0.001, (35%)	1.222±0.011^* ***** p<0.001, (39.96%)	1.106±0.007^* ***** p<0.001, (47.83%)
Aqueous ethanol extract (100 mg/kg)	1.100±0.011^* ***** p<0.001, (25.37%)	1.204±0.029^* ***** p<0.001, (35.68%)	1.150±0.029^* ***** p<0.001, (42.5%)	1.080±0.028^* ***** p<0.001, (46.85%)	1.000±0.028^* ***** p<0.001, (52.83%)
Aqueous ethanol extract (200 mg/kg)	0.998±0.066^* ***** p<0.001, (32.29%)	1.044±0.102^* ***** p<0.001, (44.23%)	0.786±0.046^* ***** p<0.001, (60.7%)	0.692±0.040^* ***** p<0.001, (65.94%)	0.562±0.010^* ***** p<0.001, (73.49%)
Butanol fraction (50 mg/kg)	1.306±0.043^ns ns non-significant (11.39%)	1.544±0.048^* ***** p<0.001, (17.52%)	1.450±0.064^* ***** p<0.001, (27.5%)	1.302±0.058^* ***** p<0.001, (35.93%)	1.220±0.051^* ***** p<0.001, (42.45%)
Butanol fraction (100 mg/kg)	1.224±0.045^* (16.96%)	1.468±0.067^* ***** p<0.001, (21.58%)	1.396±0.053^* ***** p<0.001, (30.2%)	1.326±0.042^* ***** p<0.001, (34.74%)	1.198±0.035^* ***** p<0.001, (43.49%)
Butanol fraction (200 mg/kg)	1.214±0.101^ p<0.01, (17.64%)	1.344±0.051^* ***** p<0.001, (28.21%)	1.084±0.089^* ***** p<0.001, (45.8%)	0.872±0.042^* ***** p<0.001, (57.09%)	0.712±0.039^* ***** p<0.001, (66.42%)
Aqueous Fraction (50 mg/kg)	1.254±0.062^ns ns non-significant (14.93%)	1.426±0.100^* ***** p<0.001, (23.82%)	1.350±0.078^* ***** p<0.001, (32.5%)	1.288±0.069^* ***** p<0.001, (36.61%)	1.208±0.086^* ***** p<0.001, (43.02%)
Aqueous Fraction (100 mg/kg)	1.170±0.065^* * p<0.05, (20.62%)	1.382±0.104^* ***** p<0.001, (26.18%)	1.320±0.081^* ***** p<0.001, (34%)	1.250±0.106^* ***** p<0.001, (38.48%)	1.166±0.114^* ***** p<0.001, (45%)
Aqueous Fraction (200 mg/kg)	1.150±0.079^ p<0.01, (21.98%)	1.278±0.043^* ***** p<0.001, (31.73%)	0.870±0.043^* ***** p<0.001, (56.5%)	0.788±0.070^* ***** p<0.001, (61.22%)	0.660±0.062^* ***** p<0.001, (68.87%)

Treatment groups	2^nd day	4^th day	6^th day	8^th day	10^th day
Increase in paw diameter (mm)
Arthritic control (3 mL/kg)	6.436±0.098	8.454±0.023	10.578±0.024	11.11±0.026	12.772±0.128
Piroxicam (10 mg/kg)	4.800±0.198^* ***** p<0.001, (25.42%)	5.742±0.303^* ***** p<0.001, (32.08%)	5.282±0.180^* ***** p<0.001, (50.07%)	4.102±0.094^* ***** p<0.001, (60.08%)	3.744±0.133^* ***** p<0.001, (70.69%)
Aqueous ethanol extract (50 mg/kg)	5.874±0.097^* * p<0.05, (8.73%)	7.226±0.078^* ***** p<0.001, (14.52%)	7.252±0.097^* ***** p<0.001, (31.44%)	7.222±0.106^* ***** p<0.001, (35%)	7.080±0.108^* ***** p<0.001, (44.56%)
Aqueous ethanol extract (100 mg/kg)	5.228±0.110^* ***** p<0.001, (18.76%)	5.810±0.183^* ***** p<0.001, (31.27%)	5.734±0.188^* ***** p<0.001, (45.79%)	5.644±0.177^* ***** p<0.001, (49.20%)	5.606±0.156^* ***** p<0.001, (56.11%)
Aqueous ethanol extract (200 mg/kg)	4.812±0.134^* ***** p<0.001, (25.23%)	4.808±0.161^* ***** p<0.001, (43.13%)	4.242±0.075^* ***** p<0.001, (59.89%)	3.866±0.122^* ***** p<0.001, (65.21%)	3.528±0.033^* ***** p<0.001, (72.38%)
Butanol fraction (50 mg/kg)	6.122±0.114^ns ns non-significant (4.88%)	8.216±0.272^ns ns non-significant (2.82%)	8.05±0.290^* ***** p<0.001, (23.89%)	7.584±0.328^* ***** p<0.001, (31.74%)	7.442±0.334^* ***** p<0.001, (41.73%)
Butanol fraction (100 mg/kg)	5.70±0.154^ns ns non-significant (3.76%)	7.422±0.189^ p<0.01, (12.21%)	6.872±0.293^* ***** p<0.001, (35.03%)	6.144±0.353^* ***** p<0.001, (44.70%)	5.930±0.369^* ***** p<0.001, (53.57%)
Butanol fraction (200 mg/kg)	5.504±0.131^* * p<0.05, (14.48%)	6.422±0.102^* ***** p<0.001, (24.04%)	6.550±0.119^* ***** p<0.001, (22.52%)	6.104±0.154^* ***** p<0.001, (45.06%)	5.760±0.159^* ***** p<0.001, (54.90%)
Aqueous fraction (50 mg/kg)	6.086±0.139^ns ns non-significant (5.44%)	7.092±0.144* (16.11%)	6.924±0.083^* ***** p<0.001, (34.54%)	7.002±0.164^* ***** p<0.001, (36.98%)	6.178±1.513^* ***** p<0.001, (51.63%)
Aqueous fraction (100 mg/kg)	5.248±0.068^ns ns non-significant (18.46%)	6.410±0.154^* ***** p<0.001, (24.18%)	6.506±0.112^* ***** p<0.001, (38.49%)	5.994±0.064^* ***** p<0.001, (46.05%)	5.696±0.081^* ***** p<0.001, (55.40%)
Aqueous fraction (200 mg/kg)	4.940±0.094^ p<0.01, (23.24%)	5.570±0.155^* ***** p<0.001, (34.11%)	5.600±0.173^* ***** p<0.001, (47.06%)	4.860±0.098^* ***** p<0.001, (56.26%)	4.300±0.116^* ***** p<0.001, (66.33%)

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Evaluation of in vitro and in vivo therapeutic efficacy of Ribes alpestre Decne in Rheumatoid arthritis