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. 2023 Jun 5;8(24):21696-21708.
doi: 10.1021/acsomega.3c01045. eCollection 2023 Jun 20.

Mercuric Chloride Induced Nephrotoxicity: Ameliorative Effect of Carica papaya Leaves Confirmed by Histopathology, Immunohistochemistry, and Gene Expression Studies

Yuvaraj Maria Francis  1 , Balaji Karunakaran  1 , Fauzia Ashfaq  2 , Malak Yahia Qattan  3 , Irfan Ahmad  4 , Ali G Alkhathami  4 , Mohammad Idreesh Khan  5 , Mohan Varadhan  6 , Lakshmanan Govindan  1 , Sankaran Ponnusamy Kasirajan  7
Affiliations

Mercuric Chloride Induced Nephrotoxicity: Ameliorative Effect of Carica papaya Leaves Confirmed by Histopathology, Immunohistochemistry, and Gene Expression Studies

Yuvaraj Maria Francis et al. ACS Omega. .

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Abstract

The present study analyzes the efficacy of the ethanolic extract of C. papaya leaves (ECP) against HgCl2-induced nephrotoxicity. The effects on the biochemical and percentage of body and organ weight against HgCl2-induced nephrotoxicity in female Wistar rats were studied. Wistar rats were divided into five groups with six animals in each group: control, HgCl2 (2.5 mg/kg b.w.), N-acetylcysteine (NAC 180 mg/kg) + HgCl2, ECP (300 mg/kg b.w.) + HgCl2, and ECP (600 mg/kg) + HgCl2 groups. After 28 days of study, animals were sacrificed on the 29th day to harvest the blood and kidneys for further analysis. The effect ECP was analyzed by immunohistochemistry (NGAL) and real-time PCR (KIM-1 and NGAL mRNA) in HgCl2-induced nephrotoxicity. The results revealed that the HgCl2 group showed prominent damage in the proximal tubules and glomerulus of nephrons and enormous expression of NGAL in immunohistochemistry and KIM-1 and NGAL in real-time PCR compared to the control group. The simultaneous pretreatment with NAC (180 mg/kg) and ECP (600 and 300 mg/kg) reduced renal damage and expression of NGAL in immunohistochemistry and KIM-1 and NGAL gene in real-time PCR. This study attests to the nephroprotective effect of ECP against HgCl2-induced toxicity.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Mechanism of HgCl2-induced nephrotoxicity and its prevention.
Figure 2
Figure 2
(a) DPPH free radical scavenging assay of ECP. (b) Antioxidant activity of ascorbic acid (std).
Figure 3
Figure 3
Effect of ECP at doses of 300 and 600 mg/kg compared with NAC 180 mg/kg in HgCl2-induced toxicity (2.5 mg/kg) on CK and GGT. The values are expressed as mean ± SE (n = 6 each). The F and P values are by one-way ANOVA with the Student–Newman–Keuls multiple comparison test. aSignificantly different from the control group. bSignificantly different from the HgCl2 group. cSignificantly different from the NAC group.
Figure 4
Figure 4
Effect of ECP 300 and 600 mg/kg compared with NAC 180 mg/kg in HgCl2 2.5 mg/kg toxicity on body and kidney weight, respectively, expressed in percentage. aSignificantly different from the control group. bSignificantly different from the HgCl2 group. cSignificantly different from the NAC group.
Figure 5
Figure 5
Effect of ECP at 300 and 600 mg/kg compared with NAC 180 mg/kg in HgCl2 2.5 mg/kg on OH– and GST content in kidney. Values are mean ± SE (n = 6 each).
Figure 6
Figure 6
Effect of ECP 300 and 600 mg/kg compared with NAC 180 mg/kg in HgCl2 toxicity (HgCl2, 2.5 mg/kg) on CAT, GSH, and GR; nmol of oxidized glutathione reduced/min/mg protein. Values are mean ± SE (n = 6 each). The F and P values are by one-way ANOVA with the Student–Newman–Keuls multiple comparison test. aSignificantly different from the control group. bSignificantly different from the HgCl2 group. cSignificantly different from the NAC group.
Figure 7
Figure 7
Histopathology of the kidney using PAS stain (n = 6) (100× magnification). (A) The control kidney shows well-defined glomeruli with a Bowman’s capsule and renal tubules. (B) The HgCl2 2.5 mg/kg body weight (b.w.) treated kidney shows shrunken glomeruli with a dilated Bowman’s capsule and degenerated renal tubules. (C) The NAC 180 mg/kg b.w. + HgCl2 treated kidney shows minimal renal glomeruli and tubule degeneration. (D) The ECP 300 mg/kg b.w. + HgCl2 treated kidney shows shrunken glomeruli and cell infiltration. (E) The ECP 600 mg/kg b.w. + HgCl2 treated kidney shows near-normal glomeruli with a Bowman’s capsule and renal tubules.
Figure 8
Figure 8
Histopathology of the kidney using Masson’s trichrome stain (n = 6) (10× magnification). (A) The control kidney shows well-defined glomeruli with a Bowman’s capsule and renal tubules. (B) The HgCl2 2.5 mg/kg b.w. treated kidney shows shrunken glomeruli with a dilated Bowman’s capsule and degenerated renal tubules. (C) The NAC 180 mg/kg b.w. + HgCl2 treated kidney shows minimal renal glomeruli and tubule degeneration. (D) The ECP-300 mg/kg b.w. + HgCl2 treated kidney shows shrunken glomeruli and cell infiltration. (E) The ECP 600 mg/kg b.w. + HgCl2 treated kidney shows near-normal glomeruli with a Bowman’s capsule and renal tubules.
Figure 9
Figure 9
NGAL expression in kidney, magnification 40×, arrow indicating immunopositive region. (A) Control, saline (2 mL/kg b.w.). (B) HgCl2 (2.5 mg/kg b.w.). (C) NAC (180 mg/kg b.w.). (D) ECP (300 mg/kg b.w.). (E) ECP (600 mg/kg b.w).
Figure 10
Figure 10
Graphical representation of KIM1 and NGAL expression in the kidney. aSignificantly different from the control group. bSignificantly different from the mercuric chloride group.
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