| BackgroundBenign prostatic hyperplasia (BPH) is a common urological disease among aging males. The incidence of BPH among the old men in their 50s,60s and 80s has reached 50%,70% and 92%, respectively, when aging comes, as epidemiologic data shows. Even worse, nearly half of them need clinical intervention due to lower urinary tract symptoms (LUTS), acute urinary retention (AUR), recurrent urinary tract infection or renal dysfunction caused by bladder outlet obstruction (BOO). Currently, the etiology of BPH is the core of BPH management and prevention, as well as one of the hotspot for urology and geriatric medicine, though the specific mechanism has not been fully elucidated. To date, aging and functional testes are both important basis and prerequisites for development of BPH. Recently, metabolic mechanism, hormone disturbance and chronic inflammation have been considered to be associated with the development of BPH. Nowadays drugs intervention, which through the regulation of estrogen and androgen, amelioration of LUTS and microcirculation, echancement of immunity, has become an important approach for the treatment of BPH. Traditional Chinese medicine (TCM), with its traits of safety, less adverse effect, definite therapeutic effect and suitable for long-term use, has played a certain role in the treatment of BPH, which deserves a good prospect.BPH is categorized to "Longbi ", "Jingzhuo" and "Linzheng" in TCM. It is sophisticated in the dialectical analysis of TCM, which considers as damp-heat accumulation, lung heat rise, spleen-qi fails to rise, kidney deficiency and liver-qi stagnation, etc. Xialiqi capsule, with the principle of prescription known as tonifying qi, invigorating the blood and clearing damp, has the efficacy of spleen and kidney nourishment, which is suitable for patients with BPH presented as "spleen and kidney deficiency and phlegm and blood stasis zheng".In modern biological medicine research, the use of animal disease models for pharmaceutical intervention experiment has become an important method, whicn help to conveniently and effectively understand the laws of human disease, as well as the corresponding solutions for management and prevention to be made. Therefore, this study is aiming to establish an androgen-induced BPH model of rat. Meanwhile, by using Xialiqi capsule, we observe its inhibition role on the prostate of experimental BPH model, and try to preliminarily explore the mechanism through the aspect of balance of reproductive hormones, cell proliferation and apoptosis-related proteins, growth factors, oxidative stress injuries and cytokines, in order to provide experimental evidence for its clinical application.Objective1. SD rats were subcutaneously injected with testosterone propionate after castration to establish a BPH model; Xialiqi capsule is used to explore the protective effects on prostatic histomorphology of the BPH model.2. To investigate the effects of Xialiqi capsule on the expression of dihydrotestosterone (DHT) in the serum and prostate tissue of BPH rat model.3. To explore the effects of Xialiqi capsule on the expression of proliferating cell nuclear antigen (PCNA) and caspase-3 in the prostate tissue of BPH rat model.4. To examine the effects of Xialiqi capsule on the expression of basic fibroblast growth factor (bFGF) in the prostate tissue of BPH rat model.5. To detect the effects of Xialiqi capsule on the activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) in the serum and prostate tissue of BPH rat model.6. To probe into the effects of Xialiqi capsule on the expression of interleukin 8 (IL-8) and tumor necrosis factor a (TNF-a) in the serum and prostate tissue of BPH rat model.7. From the above, we try to illustrate the efficacy and mechanism of Xialiqi capsule on BPH rat model and lay a foundation for the clinical treatment of BPH.Methods50 male SD rats were randomly divided into 5 groups,10 in each:Group A: control group (sham group); Group B:the model group; Group C:Xialiqi capsule high-dose group (1.20 g/kg·d); Group D:Xialiqi capsule low-dose group (0.61 g/kg·d); Group E:positive control group (finasteride group,0.8 mg/kg·d).Then the rats were narcotized after weighed, and bilateral testes were castrated under aseptic conditons (rats from Group A were treated with sham operation). Testosterone propionate was subcutaneously injected with a dose of 5 mg/kgd from day 8 after operation for a month (rats from Group A were treated with normal saline of equal volume). All rats were fed rountinely during modeling. After confirmation of the model by biopsy of the prostate a month later, treatments stated above were administered to the rats. All experimental rats were anaesthetised to expose the peritoneal cavity and the appearance and adhesions of prostate were observed after the total treatment was finished. Then the prostate was excised carefully and the prostate capsule was removed. A portion of the ventral prostate was fixed with formalin solution and stained with haematoxylin and eosin and followed the examination of pathology changes. Immunohistochemistry and immunofluorescence were used to detect the expression of PCNA, caspase-3 and bFGF from the prostate tissues. ELISA was used to investigate the expressions of SOD, MDA, DHT, IL-8 and TNF-a in prostate tissues.All experimental data were analyzed by SPSS 20.0 software, and presented as mean±standard deviation. After normality test and homogeneity test of variances, One-Way ANOVA was used when multiple group samples were compared. When heterogeneity of variance between those groups was shown, rank test was used instead. LSD-t test was used to compare pairwise samples. P<0.05 was considered statistically significant.Results1. The prostate weight and prostate index in each groupCompared to control group (Group A), prostate weight in model group (Group B)] increased obviously [(1326.15±60.20) mg vs. (471.38±17.96) mg, P<0.01], while decreased significantly in Group C (914.33±36.08, mg), D (1099.76±46.28, mg) and E (817.25±53.99, mg) (P<0.01), compared to Group B. Prostate weight decreased more obviously in Group C than Group D (P<0.01), while Group E decreased the most prominently (P<0.01).Compared to control group (Group A,1.06±0.06), prostate index in model group (Group B,2.89±0.18) increased obviously (P<0.01), while decreased distinctly in Group C (2.02±0.08), D (2.39±0.11) and E (1.83±0.10) (P<0.01), compared to Group B. Prostate index reduced more conspicuously in Group C than Group D (P<0.01), while Group E decreased the most distinctly (P<0.01).2. The general situation and histological changes of the prostate in each groupGeferal situation:Group A:the prostates were pink and soft without any nodule, and it was easy to separate with no adhesion. The prostates were bigger in size, which showed dark, red and nodules, even with some fester without adhesion to surrounding tissues in Group B. The general situations of the prostates in other groups (Group C, D, E) were between group A and Group B. Group C was better than Group D, while Group E improved moderately.Results of HE staining:Group A:showed structural integrity, pink secretions, without any interstitial edema, a little bit of fibrous tissue and inflammatory cell infiltration; Group B:pathological observation showed narrow glandular cavity, partly atrophy and occlusion, interstitial fibrous tissue hyperplasia, and a little bit of chronic inflammatory cell infiltrates. Group C, D and E:structure of prostates were almost intact, there was no damage in the basement membrane, hyperplasia of granular epithelium was not obvious, a moderate amount of fibrosis and inflammatory cell infiltration were observed in interstitial and glandular cavity. Group C was better than Group D, while Group E improved moderately.3. Content of DHT in the serum and prostate in each groupResults of ELISA assay showed:The levels of DHT in the serum and prostste were significantly increased in Group B [(18.04±0.62) ng/ml and (186.14±6.26) pg/mg] when compared with Group A (7.51±0.67 ng/ml; 79.32±6.06, pg/mg) (P<0.01). The levels of DHT in the serum and prostate tissue were significantly decreased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group E [(10.11±0.78) ng/ml and (100.63±5.78) pg/mg] decreased most markedly. The levels of DHT in the prostate of Group C were decreased more significantly than that of Group D (P<0.01). While there were no significant difference of DHT in the serum between Group C and Group E (P>0.05), the levels of DHT in the prostate tissue were a bit more in Group C than in Group E (P<0.05).4. Expression of PCNA and caspase-3 in the prostate among groupsResults of immunohistochemistry showed:The levels of PCNA in the prostate were significantly increased in Group B when compared with Group A (P<0.01). The levels of PCNA in the prostste were significantly decreased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group E decreased most obviously.Results of immunoflourescence showed:The levels of caspase-3 in the prostate were significantly decreased in Group B when compared with Group A (P<0.01). The levels of caspase-3 in the prostate were significantly increased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group E increased more obviously.5. Expression of bFGF in the prostate in different groupsResults of immunoflourescence showed:The level of bFGF in the prostate was significantly increased in Group B when compared with Group A (P<0.01). The level of bFGF in the prostate was significantly decreased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group E decreased more obviously.6. Activity of SOD and content of MDA in the serum and prostate in each groupResults of ELISA assay showed:The activity of SOD in the serum and prostate were significantly decreased in Group B when compared with Group A [(116.05±6.06) U/L and (0.87±0.03) U/g vs. (248.80±6.49) U/L and (2.03±0.11) U/g, P<0.01]. The activity of SOD in the serum and prostate tissue was significantly increased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group C [(218.52±7.83) U/L and (1.67±0.05) U/g] increased most markedly, followed by Group D and Group E. There were significant differences between groups (Group C, D, E, P<0.01 or P<0.05).The levels of MDA in the serum and prostate were significantly increased in Group B [(22.62±0.78) nmol/L and (231.08±9.40) pmol/g] when compared with Group A [(6.82±0.60) nmol/L and (99.73±5.97) pmol/g](P<0.01). The level of MDA in the serum and prostate tissue was significantly decreased in Group C, D and E when compared with Group B (P<0.01). Among the treatment groups, Group C [(11.40±0.51) nmol/L and (126.81±7.84) pmol/g] decreased most markedly, followed by Group D and Group E. There was no difference between Group D and E in the content of MDA in the serum. Besides that, there were significant differences between groups (Group C, D, E, P<0.01 or P<0.05).7. Content of IL-8 and TNF-a in the prostate tissue in each groupResults of ELISA assay showed:The levels of IL-8 in the serum and prostate tissues were significantly increased in Group B [(17.12±0.73) ng/ml and (174.04±5.24) ng/mg] when compared with Group A [(5.71±0.65) ng/ml and (65.81±3.52) ng/mg](P<0.01). The levels of IL-8 in the serum and prostate tissues were significantly decreased in Group C, D and E when compared with Group B (P<0.05). Among the treatment groups, Group C [(7.42±0.71) ng/ml and (89.86±6.65) ng/mg] decreased most markedly followed by Group D and Group E. There were significant differences between groups (Group C, D, E, P<0.01).The levels of TNF-a in the serum and prostate tissues were significantly increased in Group B when compared with Group A [(1.80±0.06) ng/ml and (18.26 ±0.81) pg/mg vs. (0.55±0.04) ng/ml and (7.74+0.64) pg/mg, P<0.01]]. The levels of TNF-a in the serum and prostate tissues were significantly decreased in Group C, D and E when compared with Group B (P<0.05). Among the treatment groups, Group C [(0.84±0.05) ng/ml and (9.91±0.63) pg/mg] decreased most prominently, followed by Group D and Group E. There were significant differences between groups (Group C, D, E, P<0.01).Conclusions1. BPH model of SD rats were successfully established by subcutaneously injected with testosterone propionate (5 mg/kg·d) for 4 weeks after castration.2. Xialiqi capsule can significantly reduce the prostate weight and prostate index of BPH rat model.3. Xialiqi capsule markedly inhibits the levels of DHT both in the serum and prostate.4. Xialiqi capsule improves the prostate tissue by inhibiting the expression of PCNA and upregulation of caspase-3.5. Xialiqi capsule can alleviate the proliferation of the prostate by significantly inhibit the expression of bFGF.6. Xialiqi capsule enhances the capacity of anti-oxidation by upregulating the activity of SOD and downregulating the content of MDA.7. Xialiqi capsule mitigates the inflammation of prostate by reducing the expression of IL-8 and TNF-a in the serum and prostate.In conclusion, Xialiqi capsule may treat the BPH model through the following mechanisms:directly decrease the prostate weight and prostate index; downregulating the expression og DHT; regulating the expression levels of PCNA/caspase-3 in the prostate to inhibit the proliferation; inhibiting the expression of bFGF; regulating the balance of MDA/SOD; suppressing the expression of cytokines (IL-8 and TNF-a), which may be a new option for the treatment of BPH. |
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