Department of Pulmonary and Critical Care Medicine, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430014, P.R. China.
Department of Science and Education, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430014, P.R. China.
Endocr Metab Immune Disord Drug Targets. 2024;24(6):717-724. doi: 10.2174/1871530323666230824165645.
The lung is one of the target organs of diabetes. This study aimed to probe the protective mechanism of Jiangtang Tongmai Prescription (JTTMP) against diabetic lung injury.
JTTMP-containing serum was collected, and a high glucose and high-fat diabetic cell model was established. The cells were treated with a drug-containing serum or a CAV1-associated vector. Transfection efficiency was measured by qRT-PCR and western blot, the cell proliferative capacity was tested by CCK-8 assay, and the expression of autophagosome marker LC3B was measured by immunophluorescence assay. Expression levels of the autophagy markers LC3B, p62, and Beclin-1, and the expression levels of the fibrosis markers α-SMA, FN-1, and TGF-β1 were determined by western blot, and the levels of inflammatory factors TNF-α and IL-1β in the supernatants were assessed by ELISA.
In high glucose and high fat-induced MRC-5 cells, JTTMP-containing serum impeded the abnormal cell proliferation and the expression levels of autophagy markers, fibrosis markers, as well as inflammatory factors. CAV1 expression was decreased in MRC-5 cells treated with JTTMP-containing serum. In MRC-5 cells upon transfection with the CAV1 overexpression vector and treatment with JTTMP-containing serum, increased cell proliferation, increased LC3B, p62, Beclin-1, α-SMA, FN-1, and TGF-β1, TNF-α, and IL-1β levels were found compared with cells treated with JTTMP-containing serum alone.
This study suggests that JTTMP suppresses CAV1 expression to attenuate diabetic lung injury by reducing abnormal proliferation and autophagy, and reducing levels of fibrosis and inflammation.
肺是糖尿病的靶器官之一。本研究旨在探讨降糖通脉方(JTTMP)对糖尿病肺损伤的保护机制。
收集含 JTTMP 的血清,并建立高糖高脂糖尿病细胞模型。用含药血清或 CAV1 相关载体处理细胞。通过 qRT-PCR 和 Western blot 测定转染效率,通过 CCK-8 测定细胞增殖能力,通过免疫荧光法测定自噬体标记物 LC3B 的表达。通过 Western blot 测定自噬标记物 LC3B、p62 和 Beclin-1 的表达水平,以及纤维化标记物α-SMA、FN-1 和 TGF-β1 的表达水平,并通过 ELISA 测定上清液中炎症因子 TNF-α和 IL-1β的水平。
在高糖高脂诱导的 MRC-5 细胞中,JTTMP 含血清抑制了异常细胞增殖和自噬标记物、纤维化标记物以及炎症因子的表达。JTTMP 含血清处理的 MRC-5 细胞中 CAV1 的表达减少。在转染 CAV1 过表达载体并用 JTTMP 含血清处理的 MRC-5 细胞中,与单独用 JTTMP 含血清处理的细胞相比,细胞增殖增加,LC3B、p62、Beclin-1、α-SMA、FN-1 和 TGF-β1 水平增加,TNF-α 和 IL-1β水平增加。
本研究表明,JTTMP 通过降低异常增殖和自噬、降低纤维化和炎症水平,抑制 CAV1 表达,从而减轻糖尿病肺损伤。