动物造模,慢性前列腺炎啮齿动物实验模型 z-bio 2021-12-16 1506

　　Referral: Chronic aseptic prostatitis (CAP) or chronic pelvic pain syndrome (CPPS) is the most common type of prostatitis in clinical practice. It accounts for more than 90 years and is a common disease in urology. Refers to the inflammatory state of the prostate. The incidence is about 3-16. The National Institutes of Health (NIH) divides prostatitis into four categories: acute and chronic bacterial prostatitis (type I and II), chronic prostatitis (type III) and asymptomatic prostate Inflammation (Type IV). Among them, chronic non-bacterial prostatitis (CNP) or chronic prostatitis-chronic pelvic pain syndrome (CP/CPPS) are the most common, accounting for about 90? CP/CPPS is divided into IIIA and IIIB, depending on whether there are white blood cells in the prostate sample infiltration. The etiology, pathogenesis and pathophysiology of CNP are currently diverse. A number of studies have shown that the incidence may be related to unknown microbial infections, autoimmune diseases, oxidative stress, endocrine disorders, neuropathy and social psychology. Histologically, prostatitis is characterized by multinucleated and monocyte infiltration (neutrophils, lymphocytes, macrophages and plasma cells) in the interstitial connective tissue surrounding glands or ducts. The typical clinical symptoms are frequent urination, urgency, dysuria, abnormal urination, urination or urination after defecation, and severe general pain (lumbosacral pain, lower abdominal pain, perineum, testicles, inner thighs, etc.) that can affect normal sexual function. Chronic non-bacterial prostatitis has a significant impact on the quality of life of patients. The prostates of humans and rodents are completely anatomically different. A good animal model is essential for a better understanding of CAP. Today, many human diseases other than cancer have good animal models as sources for obtaining research data and testing treatment strategies. The evaluation criteria of CP animal models are different due to different modeling methods, so the detection methods and evaluation criteria are also different.

　　Animal model: The first step in establishing a good animal model is to successfully study the etiology of prostatitis. There is no recognized standard method for prostatitis immunity in animal models. Many researchers at home and abroad use rodents as experimental animals, such as Wistar rats, SD rats, lewis rats, C57BL/6 mice, NOD mice and so on. Different animal species have different modeling methods. Experimental autoimmune prostatitis (EAP) model: EAP rat model: Currently, there is no standard method for creating an EAP animal model. Many researchers at home and abroad use rats as experimental animals. This model, provided by Pacheco-Pruil et al., used Wistar rat gonads to purify protein and injected complete Freund’s adjuvant (CFA) into the subcutaneous tissue of the same rat to induce autoimmunity. As a result, 3 out of 8 rats (showed a pro-inflammatory response 21 days after 38 MAG inoculation. Repeated examination of the same rats showed 9 out of 20 rats (45 prostate symptoms). This appeared after 30 days. This classic modeling technique is still in use today. However, the success of this method has many limitations. At present, in domestic and foreign literature, the severity of inflammation is the purified protein. According to reports, it is not only related to the concentration of the solution, but also It is related to the time, location and dose of injection. Typical injection time is 0, 15 and 30 days. Rats have undergone a total of 5 subcutaneous injections in the plantar, inguinal area, and cervix. The concentration of the purified protein solution is too high. It will cause the death of rats, and the concentration is too low to cause autoimmunity. Some people use different concentrations of protein purification to evaluate the dose-effect relationship and found that the optimal inoculation dose is 40-60mg/ml. Donadio et al., Prostatic Acid Phosphatase ( PAP) We also found that it may be the main antigen that induces autoimmune prostatitis. When rats were immunized with recombinant rat or human PAP for 4 weeks, humoral immunity was observed in mice. Rat models and cells. Not observed To traumatic T cell response (CTL) or tissue changes, although Copenhagen rats can cause CTL response and tissue-specific prostate inflammation after intravenous injection of recombinant human PAP vaccine. However, no PAP-specific antibodies have been observed, indicating that PAP Whether to induce cellular or humoral immune response may be related to the type of rat. In addition, in Wistar rats, the spleen balls from the syngeneic EAP rat model can also be used to induce prostate inflammation. In the EAP model, rat prostatitis The main antigenic substance of autoimmunity is prostate steroid binding protein (psbp), which can cause cellular and humoral immunity. This study found that 7 days after single-drug injection, single spheres began to infiltrate, and then increased rapidly to reach the maximum on day 28. The infiltrating single spheres are mainly concentrated in the interstitial space of the prostate, accompanied by hemorrhage and partial tissue fibrosis, and inflammation occurs when obese cells appear. The infiltrating cells are mainly CD4+ T cells and several types of macrophages in CD8+ T cells.

　　EAP mouse model: Keetch et al. proposed the first EAP mouse model. Using CFA, the extract from the ventral prostate leaves of the same mouse was inoculated into C57BL/6 mice at a maximum dose of 0.75 mg, supplemented with pertussis toxin. It was found that 100 C57BL/6 mice developed extensive prostatitis. It is the degree of inflammation, concentrated on the back of the prostate, and the degree of inflammation is related to the dose. Although the results of different types of SJL and A/Jj mice are not satisfactory, we found that SJL and A/J mice cause some inflammation. Thirty days after immunization, monocyte infiltration and inflammation were found. This part is concentrated near the interstitium and blood vessels. Studies have shown that the success rate of C57BL/6 mice to establish an EAP model is higher than that of other types of mice. Zhou et al. established a CP/CPP model using T2 peptides extracted from the prostate (extracted from TRPM8 in the prostate). Use T2 + CFA + AL (OH) 3. Infusion. The experimental group was injected subcutaneously (1 ml) on days 1, 14 and 28. The previous successful CP/CPP model mainly enhanced T cell accumulation and prostate antigen antibody recognition in the prostate. In this model, the enhancement of T cells and the infiltration of macrophages into the prostate stroma and glandular epithelium were observed in experimental rats, proving that the CP/CPPS model was successfully established. In addition, high levels of pro-inflammatory cytokines (IL-1β and TNF-A) and non-specific inflammatory markers (CRP) were found in the semen of CP/CPPS patients. This model is an effective model of CP/CPPS because the levels of TNF-α and IL-β in the T2 + CFA + Al (OH) 3 group significantly increase and the CRP content increases. The inflammation in the T2 + CFA + Al (OH) 3 adjuvant immunization group was more severe than that of the other groups. The mixture of T2 + CFA + Al (OH) 3 adjuvants induced experimental autoimmune prostatitis. (EAP)), the incidence rate is 100. Ivero et al. established an EAP model using OD mice. They also mixed MAG homogenate with CFA and injected subcutaneously into multiple sites. The results showed that NOD mice had obvious prostate inflammation 10 days and 20 days after vaccination. The degree of inflammatory infiltration is very large. Moderate to severe inflammatory lesions are found in the outside, dorsal, seminal vesicles and clotting glands of the prostate. We not only discovered the humoral immune response, but also the cell-mediated immune response in the inflammatory response. Further experiments in the same group showed that two injections of 1.0 mg Wistar rat MAG or 30 mg Wistar rat PSBP can induce 80-100 prostatitis in young male OD mice. Rats will become inflamed. Mice vaccinated with MAG had more edema than mice vaccinated with psbp. In short, autoimmune modeling methods are not suitable for all rodents. Choosing Wistar rats to establish an EAP model is mainly due to the spontaneous, age-dependent and histopathological changes of prostatitis, which are very similar to human chronic prostatitis in many aspects. If the injected prostaglandin purified solution concentration is 40-60mg/ml, the success rate of modeling can reach 38? Choose OD mice to establish EAP models, mainly because they are more sensitive than C57BL/6 mice and Wistar rats, and have 100 model success rate. Confirm that nodding the mouse is suitable for building an EAP model. In general, the advantage of the NOD mouse model over the EAP rat model is that the mouse species is easy to obtain, easy to cultivate, suitable for experiments that require a large amount of sample data, and some can be used. The method of immunology. Study prostatitis. Due to its immune properties, many researchers tend to choose the mouse EAP model to study the mechanism and treatment of prostatitis. The main disadvantage is that the autoantigens of the tissues that cause autoimmune prostatitis are not yet known, and NOD mice spontaneously develop diabetes and prevent prostatitis.

　　Age-related prostatitis model: rat prostatitis model: Age-related prostatitis is a commonly used model for studying autoimmune prostatitis. Previous studies have shown that rats of different races will spontaneously develop non-bacterial prostatitis at a certain age. Spontaneous prostatitis shows moderate mononuclear cell infiltration, mainly CD4 + T cells, and inflammation mainly occurs around the stroma, acinar and lateral lobes of the prostate. Studies have shown that these rats have defects in the resistance mechanism of their own prostate antigens, which are further weakened with age and changes in the physical environment, and spontaneously with age, which can lead to autoimmune prostatitis. Different types of rats have different chances of developing spontaneous autoimmune prostatitis. Vykhovanets et al. found that male Lewis rats developed spontaneous inflammation of the outer lobes of the prostate in about 12 weeks, while SD rats developed abdominal and lateral prostates in 22-24 weeks. The incidence of inflammation is low (approximately 16 elderly Wistar rats have a spontaneous CNP incidence of 27, and Copenhagen rats have a spontaneous prostatitis incidence of 88 at the 20th week.

　　Mouse prostatitis model: The possibility of spontaneous prostatitis in rats is very small, but in the late 1990s, researchers discovered that aged OD mice only spontaneously occur in the pancreas, thyroid, parathyroid and adrenal glands. On the contrary, we found that autoimmunity develops, and so does autoimmunity. prostate. It can produce spontaneous autoimmunity. Jackson et al. in male OD mice within 20 weeks. We found that spontaneous autoimmune prostatitis may develop and remain stable. NOD mice suffer from type I insulin-dependent diabetes. Prostate leukocyte antigen is 40 weeks old, 70 OD mice develop prostatitis, which has cellular immunity and humoral immunity. The incidence of 20-30 week-old male mice is 2-4 times that of 8-week-old mice. Both obtained 8-week-old nodding mice without cellular and humoral immune responses to the prostate. Studies have shown that high-titer anti-prostate antigen IgG antibodies are based on IgG2b. It has been shown that it may appear in the serum of mice with elderly male tumors. The main prostate antigen is identified as psbp. Autoimmune prostatitis in elderly male NOD mice. The development of the disease may lead to the development of spontaneous autoimmune prostatitis, and the body's resistance to prostate antigen gradually decreases with age. It has been found that spontaneous autoimmune prostatitis can passively develop abstinence in rats. Previous studies have shown that after certain types of rats reach a certain age, spontaneous inflammation of the non-bacterial prostate appears. The establishment of this model is related to the type and age of rats and mice. Lewis, Wistar’s age development, Copenhagen rats and NOD male mice are autoimmune, which provides a good model for the study of prostatitis. It is a stable and long-term maintenance model, and its pathological form is similar to clinical symptoms. It is a good pathology. Specific human CP treatment is suitable for drug research. But because the modeling time is more than three months, the time is longer, the cost is high, and the reproducibility is not ideal, so it is used less. For the study of human chronic prostatitis, it is best to use the Copenhagen rat as a model animal. For the 20-week model, the potential success rate is 88%. Hormone and castration-induced prostatitis model: rat prostatitis model: As we all know, the prostate is an organ that depends on androgens. Androgens play an important role in the growth, function and regulation of prostate diseases. This model uses estrogen and castration methods to cause abnormal hormone levels, disrupt the animal's androgen balance, and produce a non-bacterial inflammatory response to the prostate. Some researchers used 17β-estradiol in combination with ovariectomized rats, and the results showed that three different concentrations of estradiol can successfully induce prostatitis inflammation. In addition, it was found that administering 100 μg of 17β-estradiol to Wistar rats for 2 to 5 days successfully developed prostatitis. After 10 weeks of treatment with testosterone undecanoate capsules (2.0 mg/day), newborn rats aged 10-12 weeks developed severe prostatitis. Observation showed severe edema, lymphocyte and monocyte infiltration and extensive fibrosis in the two prostates of the affected prostate lobules. This shows that the method is effective 100

　　Mouse prostatitis model: Pakarainen et al., 67 LH receptor-deficient LuR-KO mice were treated with testosterone. The outstanding feature is that there are a large number of interstitial and interstitial tissue lymphocytes in the prostate. I found that I showed prostatitis . On the 21st day, an 8-week testosterone replacement therapy began to restore the gonads and accessory functions of male lur-ko mice. Overexpression of aromatase (AROM+) can lead to increased levels of endogenous estrogen in mice. AROM+ mice are normal at the beginning of the prostate. As the level of estrogen increases, mast cells and macrophages gradually appear in the prostate, and inflammation is mainly manifested by infiltration of granulocytes and T lymphocytes. The CNP model induced by estrogen is simple, rat breeds are easily available, model reagents are easily available, and the model cost is low. The pathological changes are similar to spontaneous CNP in aged rats. This rat model is very suitable for the study of human CP therapy with good pathological specificity, because age-dependent spontaneous rat CNP is pathologically similar to human CP. ...In terms of disadvantages, combined estrogen castration requires higher aseptic technique and complex surgery.

　　Chemical prostatitis model: Chemical substances are injected directly into the animal’s prostate to produce sterile CP. Currently used chemicals include glycerin, tiglium, formaldehyde croton oil and FCA. Among them, carrageenan preparations are the most commonly used, with less damage to prostate tissue, and modeling more similar to chronic inflammation. Adhakrishnan and allu used SD rats to inject 3 aginan into the prostate to establish a CNP model and test the effect of thermal and mechanical stimulation on lowering the perineal pain threshold

　　degree. They found that the inflammatory cells in model rats are mainly monocytes and lymphocytes, and chronic inflammations such as fibrous connective tissue hyperplasia, mesenchymal congestion, and edema. Chemical modeling is a model that simulates the pathophysiology of prostatitis. By directly injecting chemicals into the prostate, it causes inflammation and inevitably causes great damage to the prostate tissue. There is a certain difference between acute prostatitis and chronic pancreatitis.

　　Model indicators: Due to different model methods, the evaluation criteria and detection methods of CP animal models are different. However, most indicators are based on histology, morphology, prostate index, urodynamics, inflammatory factors, and pain.

　　Pathological index Type I index (core index): The most direct index of CP is the obvious inflammation of prostate tissue. Pathological observation can accurately determine the severity of prostatitis, including the infiltration of inflammatory cells, the appearance of inflammatory cells, tissue edema, gland necrosis, shedding and loss of glandular epithelium, and observe whether it exists with an optical microscope. The pathological classification is as follows: "0" means that most of the prostate is in a static state, the glands are atrophy, and the glandular epithelium is wrinkled. There is no secretion in the gland cavity, and the gland cavity is significantly reduced. "I" means that most of the glands in the prostate are enlarged. There is secretion in the gland cavity, but there is very little secretion, and there are a few inflammatory cells around it. "II" means that the prostate is obviously enlarged, the glandular epithelium is thin and flat, and the gland cavity is full of red secretions. "III" means that the prostate is significantly enlarged, and the glands are enlarged by 1 to 3 times. The glandular epithelium is flat and the gland cavity is filled with red secretions. The glandular cavity is obviously enlarged and scattered around the inflammatory cells. Two people randomly scored each prostate pathological section according to the above-mentioned scoring criteria.

　　Biochemical indicators (directly related indicators): The direct indicators of CP are the significant increase in white blood cells in the prostate fluid or its homogenate, and the decrease in the distribution of lecithin bodies. Prostatitis immune factors often detect IgG, IgA and IgM in prostate homogenate. Successful model establishment significantly increases IgA levels. According to neuroendocrine theory, prostatitis can cause pain and excessive secretion of prostatic fluid. The levels of NE, PG, histamine, 5-HT and NGF in the prostate homogenate can increase the levels of IL-6, IL-8, IL-10, TNF-α and IL-1β. According to immunological theory, when the model replicates normally, serum T or prostate tissue homogenate will reduce T, E2, serum prostate-specific antigen (PSA) and C-reactive protein (CRP) levels. let me.

　　Apparent indicators, urine output indicators, pain indicators: Obvious indicators include animal prostate indicators, urine output, fluid intake, and open activities. After the successful modelling, the wet prostate weight and prostate index increased, urine output decreased, drinking water decreased, open activities decreased, and hair gloss decreased. Prostatitis can cause changes in the urinary system. Urinary dynamic parameters such as bladder pressure, urination interval, and maximum urine pressure of animals with prostatitis can directly reflect the condition of prostatitis.

　　Conclusion: The cause of chronic prostatitis is complicated, the pathophysiological process is not clear, and clinical treatment is very difficult. Establishing an effective animal model is particularly important for better understanding the etiology of autoimmune prostatitis and finding the correct diagnosis and treatment methods. In practical applications, you can choose a suitable test method according to the purpose of the experiment and the display conditions. The above method still has many shortcomings. Therefore, it is necessary to continue to find a model that is more suitable for the etiology, pathogenesis and clinical symptoms of prostatitis.