Ethnopharmacological activity of <i>Hedera nepalensis</i> K. Koch extracts and lupeol against alloxan-induced type I diabetes

Hashmi, Waleed Javed; Ismail, Hammad; Jafri, Laila; Mirza, Bushra

doi:10.1590/s2175-97902019000318406

Waleed Javed Hashmi

Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan, Department of Life Sciences, Abasyn University Islamabad Campus, Islamabad, Lahore College for Women University, Lahore

Hammad Ismail

Department of Biochemistry and Biotechnology, University of Gujrat, Gujrat, Pakistan

Laila Jafri

Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan, Department of Life Sciences, Abasyn University Islamabad Campus, Islamabad, Lahore College for Women University, Lahore

Bushra Mirza ^**Correspondence: B. Mirza, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan. Telephone: +92-51-90643007, Fax: +92-51-90644050. Email: bushramirza@qau.edu.pk. These authors contributed equally in work

Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan, Department of Life Sciences, Abasyn University Islamabad Campus, Islamabad, Lahore College for Women University, Lahore

http://orcid.org/0000-0002-4134-6636

*Correspondence: B. Mirza, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan. Telephone: +92-51-90643007, Fax: +92-51-90644050. Email: bushramirza@qau.edu.pk. These authors contributed equally in work

COMPETING INTERESTS

Authors declare no conflict of interests regarding the publication of this paper.

AUTHOR’S CONTRIBUTIONS

HI and WJH conducted all the assays and experimental work. BM conceived the study design and supervised the study. LJ contributed in study design and isolated lupeol previously. HI and WJH drafted the manuscript and all authors proof read the final version.

Groups	0 h	0.5 h	1 h	2 h	4 h	3^rd day	5^th day	9^th day	12^th day	15^th day
DC	399 ± 9.3	505 ± 4.5	508 ± 3.6	496 ± 3.8	490 ± 4.0	495 ± 2.9	490 ± 4.3	492 ± 3.1	493 ± 2.0	499 ± 3.8
NC	98 ± 3.9^* * p < 0.05 and	91 ± 3.3^* * p < 0.05 and	97 ± 6.4^* * p < 0.05 and	101 ± 3.1^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	93 ± 5.5^* * p < 0.05 and	92 ± 2.8^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	95 ± 3.3^* * p < 0.05 and	92 ± 4.1^* * p < 0.05 and	94 ± 1.8^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	96 ± 4.0^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas
DC+HNC	401 ± 11.7^NS NS represents non-significant.	425 ± 13.2^NS NS represents non-significant.	399 ± 12.5^NS NS represents non-significant.	289 ± 9.1^NS NS represents non-significant.	248 ± 17.9^NS NS represents non-significant.	250 ± 4.7^* * p < 0.05 and	265 ± 5.5^* * p < 0.05 and	255 ± 3.3^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	225 ± 4.2^* * p < 0.05 and	224 ± 4.9^* * p < 0.05 and
DC+HNN	400 ± 6.19^* * p < 0.05 and	254 ± 4.7^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	201 ± 2.4^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	179 ± 4.0^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	159 ± 4.3^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	155 ± 2.5^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	149 ± 3.8^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	154 ± 2.2^* * p < 0.05 and	148 ± 2.2^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	150 ± 2.4^* * p < 0.05 and
DC+HNE	394 ± 5.3^NS NS represents non-significant.	362 ± 10.9^NS NS represents non-significant.	345 ± 6.4^* * p < 0.05 and	333 ± 9.9^NS NS represents non-significant.	343 ± 6.1^* * p < 0.05 and	348 ± 4.3^* * p < 0.05 and	340 ± 4.9^* * p < 0.05 and	342 ± 4.8^* * p < 0.05 and	335 ± 3.3^* * p < 0.05 and	343 ± 3.9^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas
DC+HNA	407 ± 4.2^NS NS represents non-significant.	455 ± 28.6^NS NS represents non-significant.	402 ± 6.7^* * p < 0.05 and	356 ± 9.1^NS NS represents non-significant.	365 ± 6.5^* * p < 0.05 and	358 ± 5.6^* * p < 0.05 and	339 ± 3.7^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	322 ± 3.6^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	346 ± 4.1^* * p < 0.05 and	365 ± 4.7^* * p < 0.05 and
DC+lupeol	389 ± 6.1^* * p < 0.05 and	225 ± 3.9^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	159 ± 4.3^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	128 ± 3.7^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	108 ± 4.6^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	106 ± 2.6^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	108 ± 2.7^* * p < 0.05 and	110 ± 2.9^* * p < 0.05 and	102 ± 2.4^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	105 ± 3.2^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas
DC+Gilb	373 ± 3.2^* * p < 0.05 and	185 ± 3.5^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	128 ± 2.1^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	109 ± 3.2^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	101 ± 3.9^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	103 ± 2.3^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	101 ± 1.4^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	102 ± 2.2^* * p < 0.05 and	101 ± 1.9^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas	99 ± 3.6^ p < 0.01 statistically significant as compared to diabetic control group expressed as means ± SEM whereas

Organ	Biomarker (Unit)	NC	DC	DC+HNC	DC+HNN	DC+HNE	DC+HNA	DC+lupeol	DC+Gilb
Liver	ALT (U/L)	22.1 ± 2.4^* * p < 0.05 and	57.3 ± 2.1^* * p < 0.05 and	35.2 ± 8.8^NS NS represents non-significant.	37.2 ± 1.6^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	31.2 ± 7.8^NS NS represents non-significant.	30.5 ± 3.5^* * p < 0.05 and	22.7 ± 0.9^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	23.6 ± 1.5
	AST (U/L)	22.5 ± 2.9^* * p < 0.05 and	73.6 ± 2.1^* * p < 0.05 and	42.6 ± 3.6^* * p < 0.05 and	36.7 ± 1.6^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	38.9 ± 3.1^* * p < 0.05 and	34.4 ± 2.1^* * p < 0.05 and	26.7 ± 1.0^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	24.2 ± 1.1
	Total bilirubin (mg/dL)	0.35 ± 0.05^* * p < 0.05 and	1.20 ± 0.04^* * p < 0.05 and	0.71 ± 0.06^* * p < 0.05 and	0.65 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	0.65 ± 0.07^* * p < 0.05 and	0.61 ± 0.05^* * p < 0.05 and	0.85 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	0.38 ± 0.01
	Total protein (G/dl)	5.3 ± 0.5^* * p < 0.05 and	8.2 ± 0.4^* * p < 0.05 and	7.1 ± 1.1^NS NS represents non-significant.	5.0 ± 0.2^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	6.9 ± 0.6^* * p < 0.05 and	6.4 ± 1.3^NS NS represents non-significant.	6.1 ± 0.3^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	6.1 ± 0.3
Kidney	BUN (mg/dL)	19.5 ± 0.8^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	35.5 ± 1.5^* * p < 0.05 and	32.2 ± 1.9^* * p < 0.05 and	25.6 ± 1.4^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	31.6 ± 1.8^* * p < 0.05 and	28.4 ± 1.1^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	23.2 ± 1.0^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	21 ± 1.2
Kidney	Creatinine (mg/dL)	0.37 ± 0.04^* * p < 0.05 and	1.61 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.22 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	0.75 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.25 ± 0.05^* * p < 0.05 and	0.92 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	0.56 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	0.5 ± 0.01

Organ	Treatment	Protein (µg/mg tissue)	TBARS (nM/min/mg tissue)	H₂O₂ (nM/min/mg tissue)	Nitrite (µM/mL)	CAT (U/min)	SOD (U/mg protein)	POD (U/min)	GSH (mM/g tissue)
Pancreas	DC	12 ± 0.41^* * p < 0.05 and	43.6 ± 0.17^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4 ± 0.04^* * p < 0.05 and	16.3 ± 0.13^* * p < 0.05 and	1.8 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.1 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.3 ± 0.07^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.1 ± 0.21^* * p < 0.05 and
	NC	10.1 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	30.4 ± 0.19^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.7 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	9.7 ± 0.13^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	6.18 ± 0.06^* * p < 0.05 and	2.6 ± 0.06^* * p < 0.05 and	5.7 ± 0.15^* * p < 0.05 and	10.2 ± 0.12^* * p < 0.05 and
	DC+HNC	11 ± 0.26^* * p < 0.05 and	44.5 ± 0.37^* * p < 0.05 and	3.5 ± 0.17^NS NS represents non-significant.	15.1 ± 0.55^* * p < 0.05 and	1.18 ± 0.08^* * p < 0.05 and	1.14 ± 0.04^* * p < 0.05 and	1.6 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.7 ± 0.13^* * p < 0.05 and
	DC+HNN	8.5 ± 0.13^* * p < 0.05 and	27.3 ± 0.15^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.4 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	13 ± 0.29^* * p < 0.05 and	7.6 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.8 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	8.2 ± 0.09^* * p < 0.05 and	7.2 ± 0.21^* * p < 0.05 and
	DC+HNE	8.6 ± 0.13^* * p < 0.05 and	33.5 ± 0.11^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.9 ± 0.07^* * p < 0.05 and	14 ± 0.31^* * p < 0.05 and	2.4 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.7 ± 0.01^* * p < 0.05 and	3.6 ± 0.28^NS NS represents non-significant.	6.9 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNA	12 ± 0.87^* * p < 0.05 and	51.4 ± 0.78^NS NS represents non-significant.	3.2 ± 0.09^* * p < 0.05 and	14.4 ± 0.86^NS NS represents non-significant.	1.17 ± 0.09^* * p < 0.05 and	1.6 ± 0.01^* * p < 0.05 and	2.6 ± 0.25^NS NS represents non-significant.	4.8 ± 0.10^* * p < 0.05 and
	DC+lupeol	8.3 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	23.3 ± 0.23^* * p < 0.05 and	2.2 ± 0.08^* * p < 0.05 and	13.6 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	8.2 ± 0.09^* * p < 0.05 and	2.1 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	10.6 ± 0.05^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	7.4 ± 0.08^* * p < 0.05 and
	DC+Gilb	10.3 ± 0.0.9	31.4 ± 0.13	3.0 ± 0.02	12.1 ± 0.12	6.5 ± 0.01	3 ± 0.04	7.2 ± 0.04	9 ± 0.01
Liver	DC	13.5 ± 0.28^* * p < 0.05 and	59.6 ± 0.19^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	5.3 ± 0.05^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	21.4 ± 0.28^* * p < 0.05 and	11.4 ± 0.12^* * p < 0.05 and	3 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.2 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4 ± 0.20^* * p < 0.05 and
	NC	11.1 ± 0.12^* * p < 0.05 and	35.1 ± 0.12^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.3 ± 0.05^* * p < 0.05 and	15.9 ± 0.34^* * p < 0.05 and	13. ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	5 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.7 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	11.7 ± 0.36^* * p < 0.05 and
	DC+HNC	13.7 ± 0.21^* * p < 0.05 and	62.07 ± 0.49^* * p < 0.05 and	4.5 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	19 ± 0.29^* * p < 0.05 and	11.7 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.5 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.6 ± 0.07^* * p < 0.05 and	4.6 ± 0.12^* * p < 0.05 and
	DC+HNN	11.7 ± 0.35^* * p < 0.05 and	43.5 ± 0.13^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.8 ± 0.11^* * p < 0.05 and	16 ± 0.29^* * p < 0.05 and	15.2 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.8 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	7.1 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	7.7 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNE	11.9 ± 0.65^NS NS represents non-significant.	50.9 ± 0.15^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.2 ± 0.02^* * p < 0.05 and	16.9 ± 0.23^* * p < 0.05 and	11.2 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.3 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	6.7 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNA	13.3 ± 0.79^NS NS represents non-significant.	53 ± 0.82^NS NS represents non-significant.	4.3 ± 0.05^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	17.3 ± 0.32^* * p < 0.05 and	6.6 ± 0.44^NS NS represents non-significant.	2.5 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.7 ± 0.10^* * p < 0.05 and	4.5 ± 0.45^* * p < 0.05 and
	DC+lupeol	11. ± 0.07^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	35.1 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.7 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	15.1 ± 0.17^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	15.7 ± 0.17^* * p < 0.05 and	5.0 ± 0.11^* * p < 0.05 and	8.3 ± 0.09^* * p < 0.05 and	8.9 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+Gilb	11.9 ± 0.06	37.3 ± 0.12	3.5 ± 0.01	16.7 ± 0.12	13.4 ± 0.06	5.2 ± 0.01	4.3 ± 0.01	10.5 ± 0.02
Kidney	DC	11.5 ± 0.07^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	60.5 ± 0.18^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.1 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	20.1 ± 0.31^* * p < 0.05 and	9.8 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.2 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.4 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.5 ± 0.15^* * p < 0.05 and
	NC	9.5 ± 0.20^* * p < 0.05 and	29.9 ± 0.49^* * p < 0.05 and	3.6 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	15.8 ± 0.19^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	12.3 ± 0.34^* * p < 0.05 and	3.7 ± 0.15^* * p < 0.05 and	4.2 ± 0.10^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	9.2 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNC	11.4 ± 0.21^* * p < 0.05 and	57 ± 0.82^* * p < 0.05 and	3.9 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	19.3 ± 0.25^* * p < 0.05 and	10.2 ± 0.11^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.3 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	1.6 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.2 ± 0.34^* * p < 0.05 and
	DC+HNN	9.8 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	29.2 ± 0.49^* * p < 0.05 and	3.3 ± 0.05^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	15.5 ± 0.13^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	13.0 ± 0.16^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.3 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.8 ± 0.05^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	8 ± 0.01^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNE	11.0 ± 0.08^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	50 ± 1.77^NS NS represents non-significant.	3.8 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	19.2 ± 0.10^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	13.2 ± 0.10^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.3 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.7 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	7.6 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+HNA	11.1 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	50 ± 0.51^* * p < 0.05 and	3.9 ± 0.06^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	17 ± 0.19^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	7.6 ± 0.09^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.1 ± 0.04^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.0 ± 0.07^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	5.2 ± 0.37^* * p < 0.05 and
	DC+lupeol	9.7 ± 0.18^* * p < 0.05 and	27.1 ± 0.13^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	3.2 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	13.5 ± 0.15^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	13.1 ± 0.11^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	2.8 ± 0.02^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	4.0 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas	8.8 ± 0.03^ p < 0.01 statistically significant as compared to positive control group expressed as means ± SEM whereas
	DC+Gilb	10.2 ± 0.08	31.1 ± 0.15	3.7 ± 0.05	16.5 ± 0.11	14.1 ± 0.18	4.3 ± 0.02	3.0 ± 0.01	8.2 ± 0.04

[1] *Correspondence: B. Mirza, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan. Telephone: +92-51-90643007, Fax: +92-51-90644050. Email: bushramirza@qau.edu.pk. These authors contributed equally in work

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Ethnopharmacological activity of Hedera nepalensis K. Koch extracts and lupeol against alloxan-induced type I diabetes