Protective effect of necrostatin1 on the damage of pancreas islet cells induced by TNFα.
10.11817/j.issn.1672-7347.2020.190534
- Author:
Bin YE
1
;
Pengfei RONG
2
;
Liang LIU
3
;
Wei WANG
2
;
Shengwang ZHANG
4
Author Information
1. Department of Radiology, Third Xiangya Hospital, Central South University, Changsha 410013. yebin9992003@163.com.
2. Department of Radiology, Third Xiangya Hospital, Central South University, Changsha 410013.
3. Department of Radiology, Yiyang Central Hospital, Yiyang Hunan 413000, China.
4. Department of Radiology, Third Xiangya Hospital, Central South University, Changsha 410013. zswang99@163.com.
- Publication Type:Journal Article
- Keywords:
TNF-α;
diabetes mellitus;
islet;
necrostatin-1
- MeSH:
Animals;
Imidazoles;
Indoles;
Insulin;
Islets of Langerhans;
Swine;
Tumor Necrosis Factor-alpha;
genetics
- From:
Journal of Central South University(Medical Sciences)
2020;45(7):752-758
- CountryChina
- Language:English
-
Abstract:
OBJECTIVES:To investigate whether necrostatin-1 (Nec-1) can protect islet cells from the damage induced by TNF-α.
METHODS:After isolation and purification, the neonatal porcine islet cell clusters (NICCs) were divided into 3 groups (islets 10 000 IEQ/group): a Nec-1 group (Nec-1+TNF-α was added to the culture medium), a TNF-α group (TNF-α was added to the culture medium), and a control group (pure medium). The number of cells was observed after 48 h of co-culture. The cell death was evaluated by AO/EB staining. Insulin secretion and DNA of islets were detected by chemiluminescence and nucleic acid quantitative analysis. RT-PCR assay was used to examine the mRNA expressions of insulin gene, glueogan gene and somatostatin gene. Flow cytometry analysis was used to detect the viability of B cells.
RESULTS:The number of islets in Nec-1 group, TNF-α group and the control group were (8 425±2 187), (4 325±778), and (7 122±1 558) IEQ, respectively. Compared to the other two groups, the number of dead cells in TNF-α group was greatly increased. The insulin/DNA values in the Nec-1 group, TNF-α group and blank control group were (13.21±3.15), (2.47±0.45), and (7.44±0.97) mIU/mg, respectively. Compared to the TNF-α group and the control group, the mRNA relative expression levels of insulin gene (6.73±1.07), glucagon gene (10.13±1.98), somatostatin gene (8.57±1.11) were significantly increased in the Nec-1 group (all <0.05), the rate of live cells (97.32±1.87)% and live B cells (90.86±3.68)% were increased significantly in the Nec-1 group (all <0.05).
CONCLUSIONS:TNF-α can induce neonatal porcine islet cells damage, which is attenuated in the presence of Nec-1. Nec-1 can increase the content of endocrine cells in NICCs.
