The Anti-Obesity Potential of Cyperus rotundus Extract Containing Piceatannol, Scirpusin A and Scirpusin B from Rhizomes: Preclinical and Clinical Evaluations

Figures & data

Figure 1 Analysis of the CRE.(A) LC-MS analysis of Cyperus rotundus extract (CRE) enriched with Piceatannol, Scirpusin A and Scirpusin B, (B) Structures of Piceatannol, Scirpusin A, and Scirpusin B.

Table 1 Demographics of Subjects in the Pilot Clinical Study

Parameter	Placebo (N=15)	CRE (N=15)
Age (Years)	37.8±11.7	37.5 ± 9.9
Height (cm)	161.8± 10.5	158.7± 9.6
Weight (Kg)	92.8 ± 21.33	88.3 ± 9.70
BMI	35.0 ± 4.66	34.7 ± 2.73
Gender	Male: 5	Male: 3
	Female:10	Female:12

Figure 2 CONSORT flow diagram of subject disposition.Eligible subjects (n = 30) were randomized into Cyperus rotundus extract (CRE) or placebo groups. By considering Last Observation Carry Forward (intent to treat analysis), 27 subjects’ data were considered for efficacy analysis. Whereas for safety analysis, 26 subjects’ data were considered.

Table 2 Effect on Body Weight and Related Parameters

Parameter	Placebo		Active		P-value
	Baseline	Visit 5	Baseline	Visit 5
Weight (Kg)	92.8 ± 21.33	92.6 ± 22.24	88.3 ± 9.70	78.8 ± 8.16*	<0.01
BMI	35.0 ± 4.66	35.1 ± 4.89	34.7 ± 2.73	31.5 ± 3.51*	<0.01
Waist circumference (cm)	106.4 ± 13.77	106.9 ± 13.93	101.5 ± 10.66	97.6 ± 9.83*	0.056
Hip circumference (cm)	125.5 ± 12.61	125.8 ± 13.02	120.9 ± 12.49	118.6 ± 11.80	0.89
Waist hip ratio	0.8 ± 0.09	0.8 ± 0.09	0.6 ± 0.06	0.8 ± 0.06	0.081
Total calorie intake (Kcal)	2423.3 ± 99.76	2482.1 ± 50.41	2413.3 ± 12.15	2463.3 ± 69.35	0.41

Note: Effect of CRE on bodyweight parameters at baseline and day 90 in participants supplemented with placebo and CRE.

Figure 3 Effect on body weight Efficacy of Cyperus rotundus extracts (CRE) in overweight subjects observed at five visits from Day 0 to day 90 compared to placebo. a) Bodyweight (Kg), b) BMI (kg/m²), and c) Waist circumference (cm). Values are represented as mean ± SE. **p < 0.01 significant difference between CRE and placebo groups on days 60 and 90.

Table 3 Effect on Lipid Levels

Serum Levels mg/dL	Placebo		Active		P-value
	Baseline	Visit 5	Baseline	Visit 5
Total cholesterol	218.5 ± 13.17	218.8 ± 14.29	219.4 ± 16.29	182.4 ± 10.27*	<0.01
Triglycerides	182.3 ± 17.28	181.8 ± 16.88	182.3 ± 30.84	171.2 ± 24.93*	0.0011
LDL- Cholesterol	168.8 ± 19.13	167.9 ± 17.78	173.5 ± 19.89	159.3 ± 19.61*	<0.01
HDL-Cholesterol	42.4 ± 2.43	42.8 ± 2.72	40.9 ± 3.70	64.4 ± 3.65*	<0.01
VLDL- Cholesterol	46.8 ± 10.19	44.4 ± 13.30	47.9 ± 11.39	33.9 ± 7.21*	0.0018

Notes: Effect of CRE on lipid levels. Lipid levels at baseline and day 90 in participants supplemented with placebo and CRE.

Abbreviations: HDL, high-density lipids; LDL, low-density lipids; TC, total cholesterol; TG, triglycerides; VLDL, very-low-density lipoproteins.

Figure 4 Anti-Adipogenesis Activity of CRE in vitro Adipogenesis inhibitory assay in 3T3-L1 mouse adipocytes. (A) Percentage inhibition of adipogenesis activity, (B) neutral triglycerides and lipids in control and treated cells stained by oil red O (ORO) (Black arrow marks), (C) fold change in the transcripts of PPARγ and aP2 genes in CRE (25 μg/mL) compared to untreated differentiated adipocytes.

Abbreviations: PPAR γ, peroxisome proliferator-activated receptor –γ; aP2, adipocyte protein 2.

Figure 5 Anti-obesity effect of CRE in High-fat diet-induced obese mice Anti-obesity effect of CRE (50, 100 and 200 mg/kg BW) in HFD-induced obesity in the C57 mice model. (A) Mean change in the body weight (g), (B) mean change in the feed consumption (g), (C) trend of mean change in body weight observed in control and treated groups from day 28 to day 55, (D) corticosterone concentration. (pg/mL), and (E) leptin concentration (pg/mL) in the high-fat diet (HFD) and CRE treated groups. Values are expressed as means ± SEM (n = 6).

Abbreviation: CRE, Cyperus rotundus extract.

Table 4 Effect of CRE Lipid, and Liver Enzymes, in C57 Mice

Group	TG (mg/dL)	TC (mg/dL)	LDL (mg/dL)	VLDL (mg/dL)	HDL (mg/dL)	ALT (mg/dL)	AST (IU/L)	Glu (mg/dL)
Control	72.29±6.5	73.61±3.9	37.9±3.7	14.79±0.82	41.53±7.1	32.41±3.9	62.46±5.2	97.52±6.9
HFD	109.5±6.2^##	122.6±7.4^##	66.16±4.7^##	25.42±1.37^##	85.48±10.3^##	39.95±2.3	86.43±8.3*	112.6±8.3
HFD+CRE50	74.05±7.9*	85.79±5.7**	50.58±5.0	17.67±1.2**	40.04±3.3**	39.32±5.2	67.52±3.5	93.4±3.7
HFD+CRE100	66.86±7.4**	88.61±3.9**	53.06±2.2	17.87±1.1**	49.92±4.8**	36.46±4.4	65.83±2.6	98.62±8.1
HFD+CRE200	68.53±9.2**	96.76±2.1**	48.44±1.8*	19.13±0.45**	21.91±3.7**	37.14±1.7	64.14±5.0*	92.3±9.1

Notes: Effect of CRE on lipid, liver enzymes, serum glucose levels in C57 mice model of HFD-induced obesity. High density lipids (HDL), Low density lipids (LDL), Total cholesterol (TC), Triglycerides (TG), Very-low-density lipoproteins (VLDL), Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Glucose (Glu), estimation in High fat diet (HFD) and CRE (50, 100 & 200 mg/kg BW) treated groups. ^##Significance (p < 0.01) compared to control; *Significance (p < 0.05) compared to HFD; **Significance (p < 0.01) compared to HFD.