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The aim of this study was to explore the protective effects of bioactive compounds from the fruit of the mulberry tree ( Morus alba  L.) against cisplatin-induced apoptosis in LLC-PK1 pig kidney epithelial cells.  Morus alba  fruit is a well-known edible fruit commonly used in traditional folk medicine. Chemical investigation of  M. alba  fruit resulted in the isolation and identification of six phytosterols ( 1 – 6 ). Their structures were determined as 7-ketositosterol ( 1 ), stigmast-4-en-3β-ol-6-one ( 2 ), (3β,6α)-stigmast-4-ene-3,6-diol ( 3 ), stigmast-4-ene-3β,6β-diol ( 4 ), 7β-hydroxysitosterol 3-O-β- d -glucoside ( 5 ), and 7α-hydroxysitosterol 3-O-β- d -glucoside ( 6 ) by analyzing their physical and spectroscopic data as well as liquid chromatography/mass spectrometry data. All compounds displayed protective effects against cisplatin-induced LLC-PK1 cell damage, improving cisplatin-induced cytotoxicity to more than 80% of the control value. Compound  1  displayed the best effect at a relatively low concentration by inhibiting the percentage of apoptotic cells following cisplatin treatment. Its molecular mechanisms were identified using Western blot assays. Treatment of LLC-PK1 cells with compound  1  decreased the upregulated phosphorylation of p38 and c-Jun N-terminal kinase (JNK) following cisplatin treatment. In addition, compound  1  significantly suppressed cleaved caspase-3 in cisplatin-induced LLC-PK1 cells. Taken together, these findings indicated that cisplatin-induced apoptosis was significantly inhibited by compound  1  in LLC-PK1 cells, thereby supporting the potential of 7-ketositosterol ( 1 ) as an adjuvant candidate for treating cisplatin-induced nephrotoxicity.

Identification of Renoprotective Phytosterols from Mulberry (Morus alba) Fruit against Cisplatin-Induced Cytotoxicity in LLC-PK1 Kidney Cells