Evaluation of in vitro and in vivo Anti-Diabetic, Anti-Hyperlipidemic and Anti-Oxidant Activity of Flower Crude Extract and Solvent Fractions of Hagenia Abyssinica (Rosaceae)

Figures & data

Table 1 Phytochemical Screening of Flower Crude Extract

No.	Secondary Metabolites	Test Results
1	Alkaloids	–
2	Saponins	+
3	Tannins	+
4	Terpenoids	+
5	Phenols	+
6	Flavonoids	+
7	Glycosides	+
8	Steroids	+
9	Anthraquinones	+

Notes: Key: (+): Indicates the presence of the constituents; (-): Indicates the absence of the constituents.

Table 2 α-Amylase Inhibitory Activities of the Crude Extract and Solvent Fractions

Concentration	Percentage (%) Inhibition
	Water Fraction	Chloroform Fraction	Crude Flower Extract	Ethyl Acetate Fraction	Acarbose
25 μg/mL	1.52±0.82	3.59±0.60	4.41±0.75	6.42±0.91	28.69±0.67
50 μg/mL	3.2±0.70	6.19±0.48	9.38±0.37	18.48±0.48	49.35±0.39
100 μg/mL	5.61±0.29	9.23±0.94	17.440.67	26.63±0.15	63.12±0.14
200 μg/mL	9.6±0.83	16.72±0.37	29.05±0.50	38.42±0.67	71.66±0.49
400 μg/mL	17.31±0.72	22.03±0.66	38.63±0.82	47.07±0.94	85.16±0.87
800 μg/mL	26.18±0.88	28.27±0.74	43.38±0.78	54.23±0.53	91.87±0.54
IC50 μg/mL	52.11±0.63	49.08±0.97	28.09±0.75	20.78±0.67	1.24±0.67

Notes: Each value of Percentage inhibition of α-amylase is presented as means ± SEM, n = 3.

Abbreviations: IC₅₀, half-maximal inhibitory concentration; SEM, standard error of mean; α-amylase, alpha-amylase.

Table 3 Antioxidant Activities of the Crude Extract and Solvent Fractions

Concentration	Percentage (%) Inhibition of DPPH
	Water	Chloroform	Ethyl Acetate	Crude Flower	Ascorbic Acid
15.625 μg/mL	2.39±0.63	3.22±0.14	6.48±0.12	6.28±0.39	14.56±0.29
31.25 μg/mL	5.08±0.29	7.44±0.81	10.48±0.55	12.64±0.27	29.04±0.67
62.5 μg/mL	9.71±0.63	11.11±0.64	13.71±0.37	17.35±0.10	51.62±0.17
125 μg/mL	18.26±0.74	22.58±0.77	31.25±0.49	33.53±0.73	59.54±0.37
250 μg/mL	22.38±0.38	33.22±0.38	39.41±0.64	42.01±0.68	72.45±0.88
500 μg/mL	36.52±0.43	48.07±0.51	51.69±0.66	58.38±0.34	89.72±0.86
IC50 μg/mL	16.96±0.13	12.14±0.64	10.78±0.54	9.41±0.14	3.59±0.94

Notes: Each value of Percentage inhibition of DPPh free radical is presented as means ± S.E.M., n = 3.

Abbreviations: DPPh, 2,2-diphenyl-1-picrylhydrazine; IC₅₀, half-maximal inhibitory concentration; SEM, standard error of mean.

Table 4 Hypoglycemic Activity of the Flower Crude Extract in Normoglycemic Mice

Group	Blood Glucose Level (mg/dl)
	0 Hours	1 Hours	2 Hours	4 Hours	6 Hours
DW10	85.50±2.16	84.50±3.40	81.67±4.34	79.00±4.59	76.50±4.09
GLC5	83.17±3.03	69.83±1.96^aA,βA	61.67±3.28^aB,βC	55.33±2.5^aC,bB,cA, βC	48.67±2.22^aC,bC,cB,βC
HAC100	78.50±3.09	76.33±2.10	74.50±2.91	71.33±3.38^nB	67.33±2.50^βA,nC
HAC200	82.00±2.82	79.33±4.26	72.83±3.55	69.00±1.51^nA, βA	63.33±1.86^aA,nB,βB
HAC400	84.17±4.00	74.50±2.77	67.83±3.22^βB	60.83±6.22^aB,βC	57.50±4.89^aC, βC

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ^acompared to the negative control, ^bcompared to HAC100, ^ccompared to HAC200, ^βcompared to baseline blood glucose level, ⁿcompared to GLC5; ^Ap < 0.05, ^Bp < 0.01,^Cp < 0.001.

Abbreviations: HAC, H. abyssinica flower crude extract; DW, distilled water; GLC, glibenclamide.

Table 5 Effect of the Flower Crude Extract in Oral Glucose Loaded Mice

Group	Blood Glucose Level (mg/dl)
	0 Minutes	30 Minutes	60 Minutes	120 Minutes
DW10	89.00±4.21	191.17±3.45^βC	148.83±6.72^βC, µB	118.67±4.88^βB,µC
GLC5	86.83±2.66	162.33±2.94^aB,bB,cA,dA,βC	75.00±6.10^aC,bC,cC,dC,µC	64.00±5.39^aC,βB, µC
HAC100	85.00±3.33	188.33±4.17^nB, βC	142.50±9.62^nC, βB, µC	90.50±8.70^µC
HAC200	87.33±4.01	184.17±6.27^nA,βC	139.33±8.08^nC, βC, µC	84.33±4.64^aA,µC
HAC400	89.67±3.52	183.50±5.14^nA, βC	131.00±4.42^nC, βC, µC	79.33±3.49^aA,µC

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ^aCompared to the negative control, ^bcompared to HAC100, ^ccompared to HAC200, ^dcompared to HAC400, ^µcompared to the blood glucose level at 30 mins, ^βcompared to baseline blood glucose level, ⁿcompared to GLC5; ^Ap < 0.05, ^Bp < 0.01,^Cp < 0.001.

Abbreviations: HAC, H. abyssinica flower crude extract; DW, distilled water; GLC, glibenclamide.

Figure 1 Effect of Hagenia abyssinica crude extract on the blood glucose level of oral glucose loaded mice.

Abbreviations: DW, distilled Water; HAC, H. abyssinica flower crude extract; GLC, glibenclamide; min, minutes.

Table 6 Antihyperglycemic Activity of a Single Dose of Flower Crude Extract and Solvent Fractions in Diabetic Mice

Group	Blood Glucose Level (mg/dl)
	0 Hours	2 Hours	4 Hours	6 Hours	8 Hours
DW10	286.67±5.44	285.83±8.20	287.00±6.06	288.67±7.15	291.17±5.42
GLC5	276.00±15.00	254.17±13.91	223.00±16.76^nA,βA	219.67±12.15^nB,βA	211.17±12.43^nC,βB
HAC100	267.83±12.66	261.33±11.61	259.17±10.95	256.83±10.55	254.50±8.14
HAC200	265.50±10.11	262.17±13.84	257.50±9.90	251.17±8.06	249.33±9.86
HAC400	279.67±8.93	274.50±14.92	267.00±8.95	256.33±9.31	244.83±10.46^nA
CFF100	262.67±17.34	262.01±13.17	258.50±11.10	257.00±14.50	255.67±13.68
CFF200	275.67±10.94	274.67±14.04	268.33±11.61	263.33±13.37	261.67±9.87
CFF400	268.50±14.76	265.67±9.74	258.50±15.19	250.67±12.41	248.17±13.24
EAF100	288.17±20.97	285.17±22.69	281.00±19.38	273.33±17.56	274.17±20.05
EAF200	274.33±14.57	269.50±11.25	258.00±13.57	257.50±14.68	250.50±15.23
EAF 400	270.33±17.07	264.50±16.78	256.00±13.35	241.50±14.38	234.17±13.56^nA
AQF100	272.83±17.39	274.00±15.90	270.50±21.41	267.50±13.94	266.67±16.07
AQF200	282.50±12.58	280.00±13.08	279.17±15.89	274.67±13.75	271.33±14.32
AQF400	265.83±9.25	263.00±11.33	259.50±14.88	252.67±9.57	252.33±8.85

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ⁿcompared to the negative control, ^βcompared to baseline blood glucose level; ^Ap < 0.05, ^Bp <0.01, ^Cp < 0.001.

Abbreviations: HAC, Hagenia abyssinica flower crude extract; DW, distilled water; CFF, chloroform fraction; EAF, ethylacetate fraction; AQF, aqueous fraction; GLC, glibenclamide.

Table 7 Antihyperglycemic Activity of Repeated Daily Doses of Flower Crude Extract and Ethylacetate Fraction in Diabetic Mice

Fasting Blood Glucose Level (mg/dl)				Percentage (%) Reduction
Group	Baseline	7th day	14th day	7th day	14th day
DC	286.67±5.44^nC	292.17±6.87^nC	294.83±5.88^nC	−1.92	−2.85
GLC5	276.00±15.00^nC	188.17±13.03^aC,βB,nC	176.83±11.75^aC, βC,nC	31.82	35.93
HAC100	267.83±12.66^nC	250.83±14.46^nC	248.17±11.38^nC	6.35	7.34
HAC200	265.50±10.11^nC	237.83±7.62^aA,nC	229.50±12.61^aA,nC	10.42	13.56
HAC400	279.67±8.93 ^nC	239.67±7.59^aA,βA,nC	221.83±9.95^aB,βC, nC	14.30	20.68
EAF100	288.17±20.97^nC	266.33±18.99^nC	259.50.33±20.19^nC	7.58	9.95
EAF200	274.33±14.57^nC	244.17±16.65^aA,nC	231.17±15.57^aA,nC	12.09	15.74
EAF400	270.33±17.07^nC	229.67±13.51^aB,βA,nC	207.83±16.85^aC, βC,nC	15.04	23.12
NC	87.33±4.43^aC	85.83±4.26^aC	89.17±3.31^aC	1.72	−2.09

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ^acompared to the diabetic control, ⁿcompared to the normal control, ^βcompared to baseline blood glucose level; ^Ap<0.05, ^Bp<0.01, and ^Cp<0.001.

Abbreviations: HAC, H. abyssinica flower crude extract; EAF, ethylacetate fraction; GLC, glibenclamide; DC, diabetic control; NC, normal control.

Table 8 Effect of Repeated Daily Doses of the Flower Crude Extract and Ethylacetate Fraction on Bodyweight of Diabetic Mice

	Body Weight (g)
Group	Before Induction of DM	Baseline	7th Day of Treatment	14th Day of Treatment
DC	27.19±0.59	26.05±0.49	24.02±0.91^nA	22.85±0.69^nC, βA
GLC5	28.06±0.35	27.66±0.33	29.04±0.46^aC,βA	30.01±0.45^aC, βA
HAC100	27.10±0.65	25.65±0.49	24.78±0.41	26.27±0.62^aB
HAC200	29.15±0.65	27.39±0.61	28.55±0.49^aB	29.48±1.26^aC, βA
HAC400	27.58±0.54	26.69±1.08	29.63±0.43^aC,βA	29.81±0.39^aC, βA
EAF100	28.04±1.32	27.79±1.08	26.91±0.88	28.40±0.72^aC
EAF200	26.70±0.74	24.89±0.83	26.58±0.77	27.19±0.71^aC
EAF400	27.54±1.15	25.89±0.82	27.81±0.76^aB	28.74±0.62^aC, βA
NC	26.47±0.89	26.59±0.66	27.40±0.71^aA	28.60±0.47^aC

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ^acompared to the diabetic control, ⁿcompared to the normal control, ^βcompared to baseline bodyweight; ^Ap<0.05, ^Bp<0.01, ^Cp<0.001.

Abbreviations: HAC, H. abyssinica flower crude extract; EAF, ethylacetate fraction; GLC, glibenclamide; DC, diabetic control; NC, normal control; DM, diabetes Mellitus.

Table 9 Effect of Repeated Daily Doses of Flower Crude Extract and Ethyl Acetate Fraction on Serum Lipid Level of Diabetic Mice

Serum Lipid Level (mg/dl)
Group	STC (mg/dl)	STG (mg/dl	HDL-c (mg/dl)	VLDLc (mg/dl)	LDL-c (mg/dl)
DC	187.17±0.79^aC	167.17±1.60^aC	23.17±2.60^aC	32.17±.79^aC	132.33±1.94^aC
GLC5	93.17±1.62^bC	88.17±1.40^bC	36.33±0.88^bC	19.83±2.49^bC	39.50±0.76^bC
HAC100	175.50±0.76^aC,bC	156.00±0.96^aC,bC	28.33±1.66^aC, bC	28.40±0.72^aC,bB	113.83±2.66^aC,bC
HAC200	167.83±1.60^aC, bC	145.67±1.66 ^aC, bC	31.83±1.60^aC, bC	26.00±1.57^aC,bC	108.67±0.61^aC,bC
HAC400	163.67±1.71^aC, bC	139.50±0.95^aC, bC	33.50±0.60^aB, bC	25.17±0.47^aC,bC	98.50±1.61^aC, bC
EAF100	175.50±3.76^aC, bC	164.33±2.42 ^aC	25.50±0.76^aC	31.33±0.88^aC	115.67±1.84^aC, bC
EAF200	172.50±2.60^aC, bC	162.50±0.76 ^aC,bA	26.50±2.76^aC, bA	29.17±0.47^aC	110.83±0.65^aC,bC
EAF400	168.33±1.14^aC,bC	156.67±0.88 ^aC,bC	29.50±1.42^aC, bC	29.00±1.36^aC,bA	103.67±1.62^aC,bC
NC	88.17±0.79 ^bC	82.67±1.11^bC	37.83±0.94^bC	18.83±0.79^bC	35.67±1.07^bC

Notes: Values are expressed as mean ± SEM (n=6 mice in each group) and analyzed by one-way ANOVA followed by Post Hoc Tuckey test; ^acompared with normal control group, ^bcompared with diabetic control; ^Ap < 0.05, ^Bp < 0.01, ^Cp < 0.001.

Abbreviations: HAC, H. abyssinica flower crude extract; EAF, ethylacetate fraction; GLC, glibenclamide; DC, diabetic control; NC, normal control; STC, serum total cholesterol; STG, serum triglyceride; HDL-c, high-density lipoprotein cholesterol; VLDL-c, very-low-density lipoprotein cholesterol; LDL-c, low-density lipoprotein cholesterol.