Indigofera oblongifolia mitigates lead-acetate-induced kidney damage and apoptosis in a rat model

Figures & data

Figure 1 HPLC–DAD phenolic profiles for the IOLE recorded at 280 nm.

Abbreviations: AU, arbitrary unit; DAD, diode array detector; HPLC, high-performance liquid chromatography; IOLE, Indigofera oblongifolia leaves extract.

Table 1 Final body and relative kidney weights in each group

Parameters	Control group	PbAc group	IOLE group	IOLE + PbAc group
Final body weight (g)	173.4±6.3	161.1±3.8	175.3±4.7	169.2±5.8b
Relative kidney weight (g/100 g bwt)	0.87±0.10	1.18±0.17a	0.97±0.11	1.01±0.08b

Notes: Values are given as mean ± SEM (n=7).

a P<0.05, significant change with respect to control;

b P<0.05, significant change with respect to PbAc for Duncan’s post hoc test.

Abbreviations: bwt, body weight; IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate; SEM, standard error of the mean.

Figure 2 Ameliorative effects of IOLE on Pb accumulation in the kidney tissue of rats treated with PbAc for 5 days.

Notes: Values are given as mean ± SEM (n=7). ^aP<0.05, significant change with respect to control; ^bP<0.05, significant change with respect to PbAc for Duncan’s post hoc test.
Abbreviations: IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate; SEM, standard error of the mean.

Figure 3 Ameliorative effects of IOLE pre-administration on ureic acid, urea, and creatinine in rats exposed to PbAc for 5 days.

Notes: Values are given as mean ± SEM (n=7). ^aP<0.05, significant change with respect to control; ^bP<0.05, significant change with respect to PbAc for Duncan’s post hoc test.
Abbreviations: IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate; SEM, standard error of the mean.

Table 2 Oxidative stress marker levels and antioxidant enzyme activities in each group

Parameters	Control group	PbAc group	IOLE group	IOLE + PbAc group
LPO (nmol/mg protein)	1.23±0.17	1.91±0.11a	1.16±0.12	1.29±0.15b
NO (µmol/mg protein)	1.68±0.12	2.23±0.16a	1.48±0.14a	1.81±0.13b
GSH (mmol/mg protein)	0.32±0.02	0.26±0.01a	0.39±0.02	0.29±0.02b
SOD (U/mg protein)	923.5±83.6	687.3±54.9a	1,068.4±98.3a	836.7±62.4b
CAT (U/mg protein)	0.41±0.02	0.29±0.02a	0.44±0.01	0.38±0.02b
GPx (µmol/mg protein)	0.68±0.03	0.51±0.03a	0.72±0.04	0.61±0.03b
GR (µmol/mg protein)	8.5±0.03	4.8±0.02a	8.9±0.05	6.8±0.03a,b

Notes: Values are given as mean ± SEM (n=7).

a P<0.05, significant change with respect to control;

b P<0.05, significant change with respect to PbAc for Duncan’s post hoc test.

Abbreviations: CAT, catalase; GPx, GSH peroxidase; GR, GSH reductase; GSH, glutathione; IOLE, Indigofera oblongifolia leaf extract; LPO, lipid peroxidation; NO, nitric oxide; PbAc, lead acetate; SEM, standard error of the mean; SOD, superoxide dismutase.

Figure 4 Ameliorative effects of IOLE pre-administration on mRNA expression of SOD2, CAT, and GPx genes in the kidney of rats exposed to PbAc for 5 days.

Notes: Values are given as mean ± SEM (n=7). ^aP<0.05, significant change with respect to control; ^bP<0.05, significant change with respect to PbAc for Duncan’s post hoc test.
Abbreviations: CAT, catalase; GPx, GSH peroxidase; GSH, glutathione; IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate; SEM, standard error of the mean; SOD, superoxide dismutase.

Figure 5 Light micrographs of the kidney.

Notes: (A) The normal histological feature of renal tubules and glomerulus in the control group. (B) Severe inflammation, extensive degeneration, cytoplasmic vacuolation, and congested glomeruli in the PbAc-treated group. (C) The normal histological structure of renal tubules and glomerulus in the IOLE group, similar to the control group. (D) The normal histological structure of renal tubules and glomerulus in the group IV, where the IOLE was pre-administered to PbAc. Some vaculated areas can be still seen, however (H&E, 400×).
Abbreviations: H&E, hematoxylin and eosin; IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate.

Figure 6 Ameliorative effects of IOLE pre-administration on mRNA expression of Bcl-2 and Bax genes in the kidney of rats exposed to PbAc for 5 days.

Notes: Values are given as mean ± SEM (n=7). ^aP<0.05, significant change with respect to control; ^bP<0.05, significant change with respect to PbAc for Duncan’s post hoc test.
Abbreviations: Bax, Bcl-2-like protein 4; Bcl-2, B-cell lymphoma 2; IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate; SEM, standard error of the mean.

Figure 7 Changes in caspase-3 expression in rat renal cells after the administration of IOLE and PbAc for 5 days.

Notes: (A) The normal histological feature of renal tubules and glomerulus in the control group with week immunostaining of caspase-3. (B) Alteration in the histological feature of renal tubules and glomerulus in the PbAc group with strong immunostaining of caspase-3. (C) The immunostaining of caspase-3 in the IOLE group, similar to that in the control group. (D) The immunostaining of caspase-3 in the group IV, where IOLE was pre-administered with PbAc, similar to that in the control group (400×).
Abbreviations: IOLE, Indigofera oblongifolia leaf extract; PbAc, lead acetate.