Effects of Invigorating the Kidney and Resolving Phlegm Chinese Herbs on Nerve-Endocrine-Immunity Network of Chronic Bronchitis Animal Model
|School||Hebei Medical University|
|Course||Chinese and Western medicine combined with the basis|
|Keywords||Chronic bronchitis Phlegm Males Androgen receptor Reproductive Endocrinology Interleukin Neuropeptide Immunohistochemistry Reverse transcription - polymerase chain reaction Histopathology|
Objective: The main purpose of this topic: an observation of the androgen receptor (AR) in male rats, mice trachea and lung expression looking for in medicine \and provide the basis for the prevention and treatment of chronic bronchitis tonifying the kidney. 2 observation of male rats caused by inhalation of cigarette smoke, and chronic bronchitis in mice model nerve - endocrine - immune network changes to investigate the pathogenesis of chronic bronchitis. The 3 observation phlegm medicine smoked chronic bronchitis animal models of nerve - endocrine - immune network, to explore the mechanism of Chinese medicine prevention and control of chronic bronchitis phlegm. METHODS: The experiment: the androgen receptor (AR) mRNA expression in normal male mouse trachea and lung tissue of 10 normal male mice, ketamine (3mg/kg taken after intraperitoneal injection) anesthesia trachea, lung and testicular Organization (all instruments after inactivated RNA enzyme treatment), quick home cryopreserved in liquid nitrogen. Different guanidine thiocyanate - phenol - chloroform, one-step extraction of total RNA and reverse transcription - polymerase chain reaction (RT-PCR) and TaqMan real-time fluorescent quantitative detection of mouse trachea, lung and testicular tissue androgen receptor ( AR) mRNA expression, ARmRNA expression levels measured directly by the ABI PRISM 7700 Sequence Detection System. The AR primer sequences: 5'-TTACAGCAGAGGCAGGAGACT-3 '5'-TGGGGCAGCTGAGTCATCCT-3' fluorescent probe sequence: 5'-FAM CCACCCTGAGAGCAGCTGCCTDABYCLE-3 'experiment: the androgen receptor (AR) in normal male rat tracheal and positioning in the lung tissue of healthy male Wistar rats were 10 ketamine (3mg/kg intraperitoneal injection) after anesthesia, cut open the chest cavity, heart, the saline perfusion induced lung completely white, taken trachea, lung and testicular set 4% paraformaldehyde for 24h, drawn into a 25% sucrose overnight soak Chinese Abstract cryostat thickness 8 11m, LABSA staining by immunohistochemistry, rat testis tissue as a positive control, with PB S instead of primary antibody as a negative control, light microscope AR expression, cytoplasmic or nuclear brown particles positive expression. HPIA Diao high resolution color pathological graphic analysis system average gray-positive cells was measured and the average luminosity, the determination of the relative content of AR in the airways and lung tissue. Experiment: cigarette smoke sucked the person law system jump chronic bronchitis animal touch type and packet administration Fangjie Kun Ming 60 healthy male mice, weighing 18 \cigarette smoke inhalation, the high-dose group of phlegm, phlegm low-dose group and Guilong Kechuanning group, in addition to the normal control group, the other four groups set smoke box clever 5cm X 50cmX 45cm), 4 / times, twice with successive 30d, then changed to 1 with continuous 30d, totaling 60d. In addition to chronic bronchitis model group, and the remaining three groups were smoked first start of the prescribed dose gavage, the normal control group and chronic bronchitis model group were given normal saline for 60d. All were killed in the 61 largest. 72 healthy male Wistar rats, weighing 180, were randomly divided into six groups, the normal control group / chronic bronchitis model group, the high-dose group of phlegm, phlegm low-dose group, bronchitis, cough Tan Chuan pill group and natural recovery group. In addition to the normal control group, the remaining five group home smoked door XI 50cm X 110cm) cigarette smoke inhalation, 64/2 times spit consecutive 38d, then changed to 1 M consecutive 38d, a total of 76d. The Chronic group when the big smoke after the end put to death. Phlegm, the low-dose group and bronchitis cough Tan Chuan pill group smoked after gavage drugs prescribed dose, normal control group and natural recovery group were given normal saline for 3 consecutive cd gavage The first cited days all killed. Experiment 4: phlegm medicine smoked chronic bronchitis in animal models histopathological taken trachea, lung tissue, fixed in 4% formaldehyde solution 24h, drawn, dehydrated in ethanol, embedded in paraffin, 4 Diao 11m slice HE staining, light microscope each group of mice and rats tracheal and lung tissue histopathological changes. Experiment 5: phlegm medicine smoked chronic bronchitis animals lemon type hypothalamic - pituitary - gonadal axis in mice Abstract blood clothing ball, rats in mg children g of ketamine door, intraperitoneal injection) anesthesia after abdominal aortic blood, centrifuged, and the serum was separated, repackaging 0 ℃ Save. By radioimmunoassay serum testosterone door * luteinizing hormone 1H into promoting follicle stimulating hormone scare SH people Chinese Abstract prolactin 0RL) content. Taken testis, epididymis weighed to calculate the viscera index. And testicular placed fixed in 4% formaldehyde solution. Derived ethanol gradient dehydration, paraffin-embedded, 4 Diao sections, HE staining. Light microscope histopathological changes in each group of mice and rat testis. Experiment 6: phlegm medicine smoked mice with chronic bronchitis model trachea, lung and testicular tissue androgen receptor uR) mRN ^ mice eyeballs rapidly after blood taken trachea, lung and testicular liquid nitrogen in cryopreservation isothiocyanate fertilizer - phenol - chloroform step extraction of total RNA, and reverse transcription - polymerase chain reaction (RTPCR) determination of the amount of mice in each group trachea, lung and testicular tissue AR mRNA expression. Experiment 7: the phlegm Herbs smoked rat chronic bronchitis model trachea, lung tissue of male hormone receptor uR) expression in rats ketamine (3mg / k, intraperitoneal injection of the airways and lungs) taken after anesthesia, 4% paraformaldehyde, dehydrated in ethanol, low melting point stone basking embedding the ,3-4urn slice, using the the immunohistochemistry LAB8A staining, HE stained, xylene, and mounted. PB S instead of primary antibody as a negative control, testicular specimens as a positive control, cytoplasm or nuclear brown particles positive fine?