Lactate dehydrogenase ldh release assay kit

A total of 60 two-month-old male SPF SD rats, weighing (180±10) g, were
purchased from Sxbex Biotechnology Company(Henan Province,China,LicenseNo.:
SCXK (豫) 2020–0005);The total flavonoid extract of D.moldevica (TFDM) is
made by the Xinjiang Uygur Autonomous Region Institute of Medicine (batch
number: 20180412, 58.4%); SDS-PAGE gel preparation kit (P0012A, Beyotime
Biotechnology Company, China), lactate dehydrogenase (LDH) release assay kit
(A020-2-2, Nanjing Jiancheng Institute of Bioengineering, China), Nitric
Oxide (NO) assay kit(A013-2-1, Nanjing Jiancheng Institute of
Bioengineering, China), Malondialdehyde (MDA) assay kit (A003-4-1, Nanjing
Jiancheng Institute of Bioengineering, China), IL-1β ELISA kit (E-EL-R0012c,
Elabscience Biotechnology, China), IL-6 ELISA kit (E-EL-R0015c, Elabscience
Biotechnology, China), TNF-α ELISA kit (E-EL-R2856c, Elabscience
Biotechnology, China), BCA protein quantification kit (23225, Thermo Fisher
Pierce, US), primary antibodies AKT(#4060, CST Company, US), p-AKT(#9272,
CST Company, US), SRC(ab40660, Abcam, US), β-actin (ab8226, Abcam, US),
GAPDH antibody(TA-08, Zhongshan Jinqiao Company, China), horseradish
enzyme-labeled goat anti-mouse IgG (ZB-2305, Zhongshan Jinqiao Company,
China), horseradish enzyme-labeled goat anti-rabbit IgG (ZB-2301, Zhongshan
Jinqiao Company, China).

Adult male Sprague-Dawley rats (clean grade, n = 117; weight, 200–250 g) were bred at Xuzhou Medical College and randomly divided into four groups: (1) sham group (n = 36), (2) I/R group (n = 36), (3) luteolin plus I/R (Lut + I/R) group (n = 36), and (4) luteolin plus the Akt inhibitor LY294002 plus I/R (Lut + LY + I/R) group (n = 9). Luteolin (purity > 98%) was purchased from Xi'an Rongsheng Biotechnology Co., Ltd. (Shanxi, China), and dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) [13 (link), 14 (link)]. LY294002 and antibodies against Akt, phospho-Akt (Thr-308, Ser-473), caspase-3, and β-actin were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Antibodies against SERCA2a, Bcl-2, and Bax were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The Ultramicro-ATPase Assay Kit and a lactate dehydrogenase (LDH) release assay kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).