Appraisal of anti-arthritic and nephroprotective potential of Cuscuta reflexa

Figures & data

Table 1. Effect of AMECR on formaldehyde induced arthritis in Sprague–Dawley rats.

	Change in paw diameter (mm)
Treatment groups	Day 2	Day 4	Day 6	Day 8	Day 10
Arthritic Control (3 mL/kg)	8.480 ± 0.046	10.417 ± 0.220	13.767 ± 0.377	16.317 ± 0.296	18.167 ± 0.300
Standard aspirin (100 mL/kg)	5.050 ± 0.050*** (40.44%)	5.667 ± 0.060*** (45.59%)	4.800 ± 0.153** (65.13%)	4.267 ± 0.093*** (73.84%)	4.120 ± 0.040*** (77.32%)
AMECR 200 mg/kg	7.167 ± 0.101*** (15.48%)	6.767 ± 0.101*** (35.03%)	5.800 ± 0.760*** (57.87%)	5.400 ± 0.870*** (66.90%)	5.250 ± 0.290*** (71.10%)
AMECR 400 mg/kg	6.550 ± 0.058*** (22.75%)	6.417 ± 0.120*** (38.39%)	5.433 ± 0.093*** (60.53%)	5.050 ± 0.050*** (69.06%)	4.867 ± 0.044*** (73.20%)
AMECR 600 mg/kg	6.250 ± 0.058*** (26.29%)	6.050 ± 0.029*** (41.92%)	5.067 ± 0.44*** (63.19%)	4.690 ± 0.097*** (71.25%)	4.225 ± 0.025*** (76.74%)

Values in the parenthesis represent percentage inhibition of paw oedema. Results are expressed as mean ± SEM (n = 5), using two way ANOVA followed by the Bonferroni post-test.

*** p < 0.001 when compared to control.

Table 2. Effect of AMECR on turpentine oil induced arthritis in Sprague–Dawley rats.

	Change in joint diameter (mm)
Treatment groups	1st hour	2nd hour	3rd hour	4th hour	5th hour	6th hour
Arthritic control	8.893 ± 0.087	13.013 ± 0.373	16.433 ± 0.262	17.917 ± 0.203	19.000 ± 0.126	21.083 ± 0.484
Standard aspirin	7.207 ± 0.012*** (18.95%)	6.843 ± 0.047*** (47.41%)	7.367 ± 0.101*** (59.95%)	6.580 ± 0.012*** (63.27%)	6.320 ± 0.012*** (66.73%)	6.173 ± 0.018*** (71%)
AMECR 200 mg/kg	7.770 ± 0.067*** (12.62%)	7.747 ± 0.039*** (40.46%)	7.543 ± 0.015*** (54.09%)	7.367 ± 0.018*** (58.88%)	7.230 ± 0.026*** (61.94%)	7.047 ± 0.032*** (66.57%)
AMECR 400 mg/kg	7.430 ± 0.015*** (16.45%)	7.327 ± 0.09*** (43.69%)	7.243 ± 0.022*** (55.92%)	7.090 ± 0.017*** (60.42%)	6.913 ± 0.019*** (63.61%)	6.743 ± 0.043*** (68.01%)
AMECR 600 mg/kg	6.993 ± 0.060*** (21.36%)	6.667 ± 0.056*** (48.76%)	6.453 ± 0.019*** (60.73%)	6.243 ± 0.022*** (65.15%)	6.120 ± 0.015*** (67.70%)	6.067 ± 0.01*** (71.22%)

Values in the parenthesis represent percentage inhibition of joint oedema. Results are expressed as mean ± SEM (n = 5), using two-way ANOVA followed by the Bonferroni post-test.

*** p < 0.001 when compared to control.

Figure 1. Effect of AMECR on percentage inhibition of protein (egg albumin) denaturation. All the values are expressed as mean ± SEM (n = 3), using two-way ANOVA followed by the Bonferroni post-test. ***(p < 0.001) vs standard group.

Figure 2. Effect of AMECR on percentage inhibition of protein (bovine serum albumin) denaturation. All the values are expressed as mean ± SEM (n = 3), using two-way ANOVA followed by the Bonferroni post-test. ***(p < 0.001) vs standard group.

Table 3. Biochemical parameters serum urea, serum creatinine, blood urea nitrogen and serum uric acid.

Biochemical Parameters (mg/dL)	Normal control	Toxic control	AMECR 200 mg/kg	AMECR 400 mg/kg	AMECR 600 mg/kg
Serum urea	43.400 ± 0.510***	61.800 ± 0.583***	48.400 ± 0.510***	44.600 ± 0.509***	41.400 ± 0.510**
Blood urea nitrogen	18.397 ± 0.088***	34.593 ± 0.547***	24.946 ± 0.659***	22.896 ± 0.257***	18.370 ± 0.328***
Serum creatinine	4.310 ± 0.073***	5.714 ± 0.126***	4.676 ± 0.068***	3.968 ± 0.034***	3.267 ± 0.076**
Serum uric acid	0.654 ± 0.021***	1.878 ± 0.048*	0.909 ± 0.013***	0.865 ± 0.017***	0.740 ± 0.032***

Values are expressed as ± SEM (n = 5), by two-way ANOVA and p<0.05 was considered as significant as compared to toxic control where

*** p < 0.001

** p < 0.01.

Table 4. Effect of Cuscuta reflexa on kidney weight of rats.

Group of Animals	Mean kidney weight (g)
Normal control	0.544 ± 0.011
Toxic control	0.702 ± 0.009
AMECR 200 mg/kg	0.624 ± 0.009
AMECR 400 mg/kg	0.606 ± 0.009
AMECR 600 mg/kg	0.586 ± 0.005

Figure 3. Histopathological analysis of the kidneys of Sprague–Dawley rats at the 8th day of treatment after gentamicin-induced nephrotoxicity. (a) Normal control group showing normal appearance of glomeruli (G) and tubules (T). (b) Toxic control group exhibiting focal necrosis (N) and lymphatics infilterate (L). (c) 200 mg/kg AMECR group presenting moderate lymphatic infilterate (L). (d) 400 mg/kg AMECR group displaying mild lymphatic infilterate (L). (e) 600 mg/kg AMECR group revealing minute lymphatic infilterate (L).

Table 5. Histopathological analysis of kidneys of gentamicin induced nephrotoxic rats.

Groups	Tubular degeneration	Tubulo-interstitial infiltration	Tubular necrosis
Normal control	Nil	Nil	Nil
Toxic control	Severe	Severe	Moderate
AMECR 200mg/kg	Moderate	Mild	Nil
AMECR 400mg/kg	Mild	Moderate	Nil
AMECR 600mg/kg	Mild	Nil	Nil

Table 6. Preliminary phytochemical analysis of Cuscuta reflexa.

Sr. No.	Name of phytoconstituents	Aqueous methanolic extract
1.	Alkaloids	++
2.	Flavonoids	+++
3.	Triterpenoids	+++
4.	Saponins	+
5.	Anthraquinones	+
6.	Tannins	–

+: Minimum presence of chemical constituents. ++: Normal presence of chemical constituents. +++: Maximum presence of chemical constituents. −: Absence of chemical constituents.

Figure 4. Peaks of phytoconstituents in HPLC analysis.

Table 7. Preliminary HPLC analysis of Cuscuta reflexa.

Compound name	Retention time	Area (mV s)	Area (%)	Quantity (Ppm)
Quercetin	2.893	20.776	0.1	1.06
p-Coumaric acid	17.280	73.255	2.8	0.94
Vanillic acid	13.007	128.1662	4.9	7.93
Gallic acid	4.160	23.380	0.9	2.82
Caffeic acid	11.887	37.435	1.4	1.78
Trans-4-hydroxy-3-methoxy cinnamic acid	24.787	302.780	11.6	1.05