Kit and detection system for evaluating aging degree of mammal and application of 8-oxoG
An 8-oxog, mammalian technology, applied in the field of evaluating the aging degree of mammals, to achieve accurate and reliable results, high sensitivity, and good specificity
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[0076] Preferably, the kit further comprises 8-oxoG with at least one element replaced by its isotope as an internal standard. According to one embodiment, the internal standard is [ 15 N 2 13 C 1 ]8-oxoG, but not limited to.
[0077]According to one embodiment, the kit is used to detect 8-oxoG in a sample through an HPLC-MS / MS system. According to this embodiment, the kit is preferably used for testing a urine sample. More preferably, the kit also includes methanol with a volume content of 5-15% and ammonium acetate with a volume content of 85-95% and a pH value of 3.5-4.0 and a concentration of 5-20mM as mobile phases of HPLC. Most preferably, the mobile phase is methanol with a volume content of 10% and ammonium acetate with a volume content of 90% and a pH value of 3.75 and a concentration of 10 mM.
[0078] Furthermore, another aspect of the present invention provides a detection system for evaluating the aging degree of mammals, said system comprising the kit of th...
Embodiment 1
[0115] Example 1: Detection of DNA and RNA oxidative damage in whole organs (tissue cells), plasma and urine of SAM mice
[0116] SAM (senescence accelerated mouse, SAM) mouse is an inbred aging model mouse obtained by Professor Takeda of Kyoto University in Japan for more than 20 years, including rapid aging mouse SAMP (senescence accelerated prone mouse, SAMP) and normal aging Two strains of mouse SAMR1 (senescence accelerated resistant mouse, SAMR). SAMP8 is a subline of the SAMP line, mainly characterized by learning and memory impairment, and its lifespan is about 12.1 months. It is currently recognized as an ideal model of natural aging dementia; SAMR shows normal aging, and is generally used as a normal model of SAMP. control.
[0117] Both SAMP8 and SAMR1 mice used in the present invention were provided by the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine. Six 1-, 4-, 8-, and 12-month-old mice were selected, and SAMR1 mice were used ...
Embodiment 2
[0127] Example 2: Detection of 8-oxodG and 8-oxoG in natural aging SD rat tissue, blood and urine
[0128] The kit, detection system and method of the present invention are used to detect natural aging SD rats without any treatment. SD rats were purchased from Beijing Weitong Lihua. Raised in an SPF environment, at 1, 3, 9, 12, 16 and 30 months of age, 6 animals were randomly selected and raised in a single cage to collect urine; after anesthesia, blood was collected from the inferior vena cava; heart, lung, liver, and kidney were collected , brain, and testis were frozen in liquid nitrogen. All animals were operated in accordance with the experimental animal operating procedures in Rockefeller University Animal Care and Use Committee with Protocol Numbers 00111 and 03017.
[0129] 8-oxodG and 8-oxoG in rat organ tissue, plasma and urine were detected, and it was found that the content of 8-oxodG and 8-oxoG increased with age, and the change trend of urine (such as Figure ...
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