Malt extract regulating the NLRP3/Caspase-1/IL-1β pathway to inhibit the proliferation and prolactin secretion of anterior pituitary cells in hyperprolactinemia rats

doi:10.11958/20221738

Abstract

Abstract:

Objective To investigate effects of malt extract (ME) on the proliferation and secretion of prolactin (PRL) in anterior pituitary cells of hyperprolactinemia (HPRL) rats by regulating the Nod-like receptor protein 3(NLRP3)/ cysteine aspartic specific protease-1 (Caspase-1)/ interleukin-1β (IL-1β) pathway. Methods The anterior pituitary cells of normal and HPRL rats were separated and named as the NC group and the model group. The expression levels of growth hormone and PRL in cells were identified by immunohistochemistry. Cells in the model group were treated with 0 mg/L ME, 25 mg/L ME, 50 mg/L ME, 100 mg/L ME, 5 mmol/L Adenosine 5'-triphosphate (ATP), 100 mg/L ME and 5 mmol/L ATP for 48 h, and named as the Blank group, the ME low dose group (ME-L group), the ME medium dose group (ME-M group), the ME high dose group (ME-H group), the ATP (NLRP3 activator) group and ME-H+ATP group. The morphology of cells in each group was observed by light microscope. Cell proliferation was detected by CCK-8 method. Cell apoptosis was detected by flow cytometry. The level of PRL in the supernatant of rat anterior pituitary cells was detected by ELISA. Western blot assay was applied to detect expression levels of proliferating cell nuclear antigen (PCNA), dopamine receptor D2 (DRD2), dopamine transporter (DAT), NLRP3, Caspase-1 and IL-1β in rat anterior pituitary cells. Results Rat anterior pituitary cells were isolated successfully. Compared with the NC group, the volume of anterior pituitary cells was smaller and the shape was irregular, the OD₄₅₀ value, prolactin level, DAT, PCNA, NLRP3, Caspase-1, IL-1β protein expression increased, and the apoptosis rate and DRD2 protein expression decreased in the blank group (P<0.05). Compared with the blank group, the morphology of anterior pituitary cells was improved in the ME-L group, the ME-M group and the ME-H group. The OD₄₅₀ value, prolactin level, DAT, PCNA, NLRP3, Caspase-1 and IL-1β protein expression levels decreased, and the apoptosis rate and DRD2 protein expression increased in a dose-dependent manner. The change trend of the corresponding indexes was contrary to the above in the ATP group (P<0.05). ATP attenuated the inhibitory effect of high-dose ME on the proliferation of anterior pituitary cells and PRL secretion in HPRL rats. Conclusion ME may inhibit the proliferation and PRL secretion of HPRL rat anterior pituitary cells by down-regulating the NLRP3/Caspase-1/IL-1β pathway.

Key words: hyperprolactinemia, NLR family, pyrin domain-containing 3 protein, Caspase 1, interleukin-1beta, receptors, dopamine D2, malt extract

CLC Number:

R584.1

Figures/Tables 8

References 19

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组别	OD₄₅₀值	细胞凋亡率（%）	PRL（mg/L）
NC组	0.39±0.03	12.64±0.31	193.36±4.12
Blank组	1.18±0.11^a	4.05±0.10^a	368.82±7.11^a
ME-L组	0.95±0.08^b	6.21±0.11^b	321.26±5.79^b
ME-M组	0.74±0.05^bc	8.36±0.12^bc	280.65±5.33^bc
ME-H组	0.45±0.04^bcd	11.12±0.13^bcd	221.27±4.34^bcd
ATP组	1.44±0.12^b	1.59±0.08^b	473.54±8.32^b
ME-H+ATP组	0.81±0.07^e	7.34±0.16^e	305.56±6.71^e
F	139.400^＊＊	3 416.000^＊＊	1 405.000^＊＊

组别	OD₄₅₀值	细胞凋亡率（%）	PRL（mg/L）
NC组	0.39±0.03	12.64±0.31	193.36±4.12
Blank组	1.18±0.11^a	4.05±0.10^a	368.82±7.11^a
ME-L组	0.95±0.08^b	6.21±0.11^b	321.26±5.79^b
ME-M组	0.74±0.05^bc	8.36±0.12^bc	280.65±5.33^bc
ME-H组	0.45±0.04^bcd	11.12±0.13^bcd	221.27±4.34^bcd
ATP组	1.44±0.12^b	1.59±0.08^b	473.54±8.32^b
ME-H+ATP组	0.81±0.07^e	7.34±0.16^e	305.56±6.71^e
F	139.400^＊＊	3 416.000^＊＊	1 405.000^＊＊

组别	DRD2	DAT
NC组	0.83±0.07	0.54±0.04
Blank组	0.27±0.02^a	1.26±0.11^a
ME-L组	0.39±0.03^b	1.07±0.09^b
ME-M组	0.51±0.04^bc	0.83±0.07^bc
ME-H组	0.75±0.06^bcd	0.67±0.05^bcd
ATP组	0.11±0.01^b	1.62±0.13^b
ME-H+ATP组	0.43±0.03^e	0.92±0.06^e
F	218.600^＊＊	114.300^＊＊

组别	DRD2	DAT
NC组	0.83±0.07	0.54±0.04
Blank组	0.27±0.02^a	1.26±0.11^a
ME-L组	0.39±0.03^b	1.07±0.09^b
ME-M组	0.51±0.04^bc	0.83±0.07^bc
ME-H组	0.75±0.06^bcd	0.67±0.05^bcd
ATP组	0.11±0.01^b	1.62±0.13^b
ME-H+ATP组	0.43±0.03^e	0.92±0.06^e
F	218.600^＊＊	114.300^＊＊

组别	PCNA	NLRP3	Caspase-1	IL-1β
NC组	0.27±0.02	0.58±0.04	0.38±0.03	0.15±0.01
Blank组	0.88±0.07^a	1.57±0.12^a	1.42±0.11^a	1.23±0.11^a
ME-L组	0.73±0.06^b	1.32±0.10^b	1.24±0.12^b	1.06±0.11^b
ME-M组	0.55±0.05^bc	1.11±0.09^bc	1.01±0.08^bc	0.83±0.09^bc
ME-H组	0.36±0.02^bcd	0.73±0.06^bcd	0.54±0.04^bcd	0.32±0.03^bcd
ATP组	1.28±0.11^b	1.82±0.13^b	1.68±0.12^b	1.58±0.11^b
ME-H+ATP组	0.63±0.05^e	1.20±0.11^e	1.12±0.13^e	1.20±0.13^e
F	183.500^＊＊	120.500^＊＊	134.900^＊＊	178.700^＊＊