Therapeutic Efficacy of Helianthemum lippii Extract and Silver Nanoparticles Synthesized from the Extract against Cadmium-Induced Renal Nephrotoxicity in Wistar Rats

Abstract

This study explored the therapeutic efficacy of Helianthemum lippii and silver nanoparticles (Ag NPs) synthesized using a H. lippii extract to alleviate cadmium-induced nephrotoxicity in Wistar rats. Sub-acute toxicity assessments of H. lippii (100 mg/kg, 1000 mg/kg, and 4000 mg/kg) and Ag NPs (2 mg/kg and 10 mg/kg) did not find any significant difference, compared with untreated control rats (n = 3 animals/group). Then, the adult Wistar rats were divided into one control (untreated/unexposed) and six experimental groups (n = 5/group): Ag NPs alone, H. lippii alone, exposure to 50 mg/kg CdCl₂ in drinking water for 35 days, exposure to CdCl₂ for 35 days followed by treatment with 0.1 mg/kg/day Ag NPs (intraperitoneal injection) and/or 100 mg/kg/day H. lippii by gavage for 15 days. In the CdCl₂-exposed group, body weight decreased; urea, creatinine, and uric acid concentrations increased (p < 0.05 vs. control), indicative of nephrotoxicity, antioxidant defenses (SOD, GSH, and CAT) were reduced, and malondialdehyde concentration increased. Moreover, the kidney's architecture in CdCl₂-exposed rats was altered: fibrosis, inflammatory cell infiltration, glomerular destruction, and tubular dilatation. Treatment with H. lippii and/or Ag NPs after CdCl₂ exposure improved some of the renal function and architecture alterations induced by CdCl₂, and also increased body weight. This study underscores the potential therapeutic applications of H. lippii and Ag NPs to decrease oxidative stress and promote xenobiotic detoxification, in line with the growing emphasis on environmentally conscious practices in scientific research and healthcare.

Keywords: Helianthemum lippii; Wistar rats; biochemical parameters; cadmium toxicity; gavage; histopathology; intraperitoneal injection; nephrotoxicity; oxidative stress; silver nanoparticles.

Figure 1

HPLC chromatogram of the H. lippii aqueous extract: 1: gallic acid; 2 chlorogenic acid; 3: caffeic acid; 4: p-Coumaric acid; 5: naringin; 6: quercetin.

Figure 2

Characteristics of synthesized Ag NPs: (a) crystalline structure (XRD data), (b) morphology (SEM image), and (c) size distribution analysis.

Figure 3

Comparison of baseline body weight (a), body weight gain (b), and relative kidney weight (c) in the seven rat groups. Group 1 (Control): no exposure/treatment. Group 2 (Cd): addition of CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day, by gavage, for 35 days). Group 4 (Cd + Hl): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, intraperitoneal injection) for 35 days. Group 6 (Cd + Ag): CdCl₂ exposure (like in Group 2) followed by Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd+ Hl +Ag): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) and Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. * p < 0.05, *** p < 0.001 vs. Group 1; a p < 0.05, b p < 0.01, c p < 0.001: vs. Group 2.

Figure 4

Comparison of different serum markers in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): exposure to CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day) by gavage for 35 days. Group 4 (Cd + Hl): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, intraperitoneal injection) for 35 days. Group 6 (Cd + Ag): CdCl₂ exposure (like in Group 2) followed by Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) and Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. (a) serum triglycerides, (b) serum cholesterol, (c) serum glucose, (d) serum albumin. * p < 0,05, ** p < 0.01, *** p < 0.001 vs. Group 1; a p < 0.05, c p < 0.001: vs. Group 2.

Figure 4

Comparison of different serum markers in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): exposure to CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day) by gavage for 35 days. Group 4 (Cd + Hl): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, intraperitoneal injection) for 35 days. Group 6 (Cd + Ag): CdCl₂ exposure (like in Group 2) followed by Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) and Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. (a) serum triglycerides, (b) serum cholesterol, (c) serum glucose, (d) serum albumin. * p < 0,05, ** p < 0.01, *** p < 0.001 vs. Group 1; a p < 0.05, c p < 0.001: vs. Group 2.

Figure 5

Renal function marker levels in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): addition of CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day), by gavage for 35 days. Group 4 (Cd + Hl): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, intraperitoneal injection) for 35 days. Group 6 (Cd + Ag)/: CdCl₂ exposure (like in Group 2) followed by Ag NPs (0.1 mg/kg, body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg, body weight/day by gavage) and Ag NPs (0.1 mg/kg, body weight/day by intraperitoneal injection) for 15 days. (a) serum urea, (b) serum creatinine, (c) serum uric acid, (d) serum blood ration urea/creatinine. * p < 0.05, *** p < 0.001 vs. Group 1; a p < 0.05, b p < 0.01, c p < 0.001: vs. Group 2.

Figure 5

Renal function marker levels in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): addition of CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day), by gavage for 35 days. Group 4 (Cd + Hl): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, intraperitoneal injection) for 35 days. Group 6 (Cd + Ag)/: CdCl₂ exposure (like in Group 2) followed by Ag NPs (0.1 mg/kg, body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group 2) followed by H. lippii extract (100 mg/kg, body weight/day by gavage) and Ag NPs (0.1 mg/kg, body weight/day by intraperitoneal injection) for 15 days. (a) serum urea, (b) serum creatinine, (c) serum uric acid, (d) serum blood ration urea/creatinine. * p < 0.05, *** p < 0.001 vs. Group 1; a p < 0.05, b p < 0.01, c p < 0.001: vs. Group 2.

Figure 6

Oxidative stress markers in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): addition of CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day, by gavage for 35 days). Group 4 (Cd + Hl): CdCl₂ exposure (like in Group II) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, by intraperitoneal injection) for 35 days. Group 6 (Cd + Ag): CdCl₂ exposure (like in Group II) followed by Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group II) followed by H. lippii extract (100 mg/kg body weight/day by gavage) and Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. (a) MDA, (b) SOD, (c) GSH, (d) Catalase. * p <0.05, *** p < 0.001 vs. Group 1; a p < 0.05, b p < 0.01, c p < 0.001: vs. Group 2.

Figure 6

Oxidative stress markers in the seven rat groups. Group 1 (Control): no treatment/exposure. Group 2 (Cd): addition of CdCl₂ (50 mg/kg body weight/day) in drinking water for 35 days. Group 3 (Hl): H. lippii extract (100 mg/kg body weight/day, by gavage for 35 days). Group 4 (Cd + Hl): CdCl₂ exposure (like in Group II) followed by H. lippii extract (100 mg/kg body weight/day by gavage) for 15 days. Group 5 (Ag): Ag NPs (100 μg/kg body weight/day, by intraperitoneal injection) for 35 days. Group 6 (Cd + Ag): CdCl₂ exposure (like in Group II) followed by Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. Group 7 (Cd + Hl + Ag): CdCl₂ exposure (like in Group II) followed by H. lippii extract (100 mg/kg body weight/day by gavage) and Ag NPs (0.1 mg/kg body weight/day by intraperitoneal injection) for 15 days. (a) MDA, (b) SOD, (c) GSH, (d) Catalase. * p <0.05, *** p < 0.001 vs. Group 1; a p < 0.05, b p < 0.01, c p < 0.001: vs. Group 2.

Figure 7

Micrographs of representative kidney tissue sections from rats in the different experimental groups, showing the effect of CdCl₂ exposure and the mitigating effect of H. lippii and Ag NPs. (A) Group 1 (control); (B) Group 2 (CdCl₂ exposure); (C) Group 3 (H. lippii extract alone); (D) Group 4 (H. lippii after CdCl₂ exposure); (E) Group 5 (Ag NPs); (F) Group 6 (Ag NPs after CdCl₂ exposure); (G) Group 7 (H. lippii and Ag NPs after CdCl₂ exposure). N, normal cell; IF, inflammatory cells; DG, destroyed glomeruli. TD, Tubular dilatation; 40×.