Effect of Andrographis paniculata polysaccharide on human retinoblastoma Y79 cell proliferation and apoptosis
Authors:   Department of Ophthalmology, Fuling Central Hospital, Chongqing 408000, China; Clinical Medicine, Dali University, Dali 671000, Yunnan Province, China, Xu Bing , Zhang Wei ,   Department of Ophthalmology, Fuling Central Hospital, Chongqing 408000, China, Li Yue ,   Department of Pathology, Fuling Central Hospital, Chongqing 408000, China, Wang Mao-Ren ,   Clinical Medicine, Dali University, Dali 671000, Yunnan Province, China, Liu Jing-Chen ,   Clinical Medicine, Dali University, Dali 671000, Yunnan Province, China, Dong Kai-Ye ,   Department of Ophthalmology, the First Affiliated Hospital, Dali University, Dali 671000, Yunnan Province, China, Fabian Ido Didi ,   Goldschleger Eye Institute, Sheba Medical Center, Tel Hashomer, Tel-Aviv University, Tel-Aviv 52621, Israel, Qiu Dong ,   Clinical Medicine, Dali University, Dali 671000, Yunnan Province, China, Li Cai-Rui ,   Department of Ophthalmology, the First Affiliated Hospital, Dali University, Dali 671000, Yunnan Province, China, Xiang Yi-Min ,   Department of Ophthalmology, Fuling Central Hospital, Chongqing 408000, China
Published: March 2021

AIM: To explore the effect of the Andrographis paniculata (A. paniculata) polysaccharide on the proliferation and apoptosis of human retinoblastoma (RB) Y79 cells and its mechanism.
METHODS: The refined A. paniculata polysaccharide was obtained using techniques such as water extraction, ethanol precipitation, and decompression concentration. The inhibition effect of the A. paniculata polysaccharide on the proliferation of Y79 cells was detected by cell proliferation assay. Flow cytometry was used for the detection of cell apoptosis rate and cycle change. Real-time qunatitative polymerase chain reaction (RT qPCR)and Western blotting were used to detect the expression of cell apoptosis signal pathway-related factors (caspase-3, caspase-8, and caspase-9) and cell cycle signal pathway-related factors (CDK1 and cyclinB1) at the transcriptional and translational levels.
RESULTS: Infrared and ultraviolet spectrum scanning showed that the extracted drug was a polysaccharide with high purity. After being treated with different concentrations of A. paniculata polysaccharide for different periods of time, the Y79 cells showed different degrees of proliferation inhibition. Flow cytometric observations showed that the cell apoptosis rate and the proportion of cells blocked in the G2/M phase were significantly increased after A. paniculata polysaccharide treatment. Further analysis revealed that the mRNA and protein expression of caspase-3, caspase-8, and caspase-9 in the A. paniculata polysaccharide treatment groups increased significantly compared with that in the control groups, while the expression of CDK1 and cyclinB1 decreased significantly.
CONCLUSION: The A. paniculata polysaccharide could inhibit the proliferation and induce apoptosis of Y79 cells. Its possible mechanism is via the upregulation of caspase-3, caspase-8, and caspase-9 expression in the cell apoptotic signaling pathway and the downregulation of CDK1 and cyclinB1 expression in the cell cycle signaling pathway.