Effect of chloroquine on type Ⅱ alveolar cell injury caused by lipopolysaccharide

doi:10.16138/j.1673-6087.2021.03.012

Abstract

Abstract:

Objective To explore the effect of chloroquine (CQ) on lipopolysaccharide (LPS)-induced damage to alveolar type Ⅱ(AT2) cell. Methods Different concentrations of LPS and CQ were used to treat AT2 cells separately, and cell counting kit-8(CCK-8) was used to detect cell viability to screen the concentration of LPS and CQ. After that, the AT2 cells were divided into 4 groups based on different treatment, including control (without treatment), LPS, CQ and LPS+CQ groups. The expression of cleaved caspase 3, Bcl-2, Bax and the phosphorylation level of Akt were detected by Western blotting after each group of cells were processed. After that, LY294002, the signaling pathway inhibitor of phosphatidylinositol-3-kinase(PI3K)/Akt, was added in the cells which were treated by LPS and CQ successively(LPS+CQ+LY294002 group), and the protein expression of cleaved caspase 3, Bcl-2 and Bax was detected by Western blotting in this group. Results The LPS in the concentration of 1, 10 and 50 mg/L was kept for 24 h. The activity of AT2 decreased significantly (P<0.05) when the LPS concentration was increased to 50 mg/L, so LPS in this concentration was used to stimulate AT2. AT2 cell viability did not change significantly (P>0.05) when they were treated with 0.5 μmol/L and 5 μmol/L CQ for 4 h. However, AT2 cell viability began to decrease and showed a concentration-dependent manner when the CQ concentration increased to 10 μmol/L. According to the cell response to CQ in different concentration, 5 μmol/L CQ was chosen to treat the cells. After stimulation with 50 mg/L LPS, the expression of cleaved caspase 3 in the cell increased, and the Bcl-2/Bax ratio and the phosphorylation of Akt decreased. After adding 5 μmol/L CQ, the cleaved caspase 3 dropped, and Bcl-2/Bax ratio and Akt protein phosphorylation rise significantly. After the PI3K inhibitor LY294002 was added, the effect of CQ was eliminated, which showed the Bcl-2/Bax ratio decreased, and the expression of cleaved caspase 3 increased. Conclusions CQ(5 μmol/L) could inhibit cell mitochondrial apoptosis by activating the PI3K/Akt signaling pathway, and alleviate the damage of AT2 cells induced by LPS.

Key words: Chloroquine, Alveolar type Ⅱ cells, Apoptosis, Phosphatidylinositol-3-kinase/Akt

CLC Number:

R631

ZHU Yingying, ZHANG Jiaojiao, SUN Junnan, WANG Hairong. Effect of chloroquine on type Ⅱ alveolar cell injury caused by lipopolysaccharide[J]. Journal of Internal Medicine Concepts & Practice, 2021, 16(03): 197-201.

Figures/Tables 5

References 18

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组别	裂解的胱天蛋白酶3	Bcl-2/Bax	p-Akt/Akt
正常对照组	0.97±0.05	1.01±0.11	0.86±0.05
LPS处理组	1.72±0.18¹⁾	0.63±0.06^1）	0.70±0.03^1）
CQ处理组	0.98±0.11	0.85±0.05	0.93±0.11
LPS+CQ组	0.99±0.04²⁾	1.10±0.10^2）	1.08±0.08^2）

组别	裂解的胱天蛋白酶3	Bcl-2/Bax
正常对照组	0.45±0.05	0.97±0.07
LPS处理组	1.13±0.10^1）	0.73±0.04^1）
LPS+CQ组	0.71±0.04^2）	1.30±0.05^2）
LPS+CQ+LY294002组	1.09±0.04^3）	0.98±0.06^3）