麝香配伍乳香对大鼠前列腺上皮细胞紧密连接结构相关蛋白表达的影响

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目的:探讨麝香-乳香配伍处理对正常及慢性前列腺炎模型大鼠前列腺上皮细胞紧密连接结构相关蛋白表达的影响。方法:80只雄性SD大鼠随机分为8组:正常空白对照组、正常麝香-乳香组、正常麝香组、正常乳香组、模型空白组、模型麝香-乳香组、模型麝香组、模型乳香组。制备慢性前列腺炎大鼠模型,造模60 d时根据组别分别给予麝香0.021 g/(kg·d)、乳香1.05 g/(kg·d)剂量灌胃,麝香-乳香组给予联合剂量,空白组给予生理盐水灌胃。各组别连续灌胃3 d,处死大鼠,取前列腺组织,固定后以免疫组化法检测大鼠前列腺上皮屏障紧密连接功能相关蛋白紧密连接蛋白1、紧密连接蛋白3、闭锁蛋白、胞质附着蛋白1(ZO-1)的表达情况。结果:在病理状态下,仅紧密连接蛋白1的表达增高有统计学意义。麝香、乳香处理后,在生理/病理状态下对4种蛋白的调节作用不同:生理状态下,单用麝香(824.6±393.3)、乳香(982.0±334.0)或配伍处理(1 088.1±640.2)均可大幅度增高紧密连接蛋白1的表达(P<0.05,P<0.01);对于紧密连接蛋白3,单用乳香表达上调(1 009.5±243.6,P<0.05),单用麝香(597.5±80.7)差异无统计学意义,配伍麝香(678.4±255.1)可降低乳香上调表达作用;单用麝香(693.0±424.8)、乳香(732.1±302.0)或配伍处理(560.2±202.3)对闭锁蛋白表达影响差异无统计学意义;单用麝香(290.0±166.8)、乳香(419.7±108.1)对ZO-1表达影响差异无统计学意义,配伍处理后表达明显下调(197.7±98.2,P<0.05)。慢性前列腺炎病理状态下,大鼠前列腺组织紧密连接蛋白1(823.0±100.1)、闭锁蛋白(1 160.0±32.2)表达显著增高(P<0.01,P<0.05);单用麝香(764.9±179.0)、乳香(468.4±220.4)或配伍(335.1±204.0)使用可下调紧密连接蛋白1表达(P<0.05);单用麝香(700.1±223.7)或乳香(744.6±94.5)可上调紧密连接蛋白3表达(P<0.05);单用麝香(749.6±321.7)、乳香(615.0±221.0)或配伍处理(505.8±523.7)均可下调闭锁蛋白表达;单用乳香可上调ZO-1表达(443.2±144.9),与麝香配伍处理后表达下调(213.5±24.9),与单用乳香组比较差异有统计学意义(P<0.05)。结论:麝香、乳香对生理/病理状态下前列腺上皮屏障结构紧密连接相关蛋白的调节作用具有选择性和双向性,主要通过ZO-1蛋白的下调实现促通透性作用,而通过调节紧密连接蛋白1、紧密连接蛋白3、闭锁蛋白表达以维持结构稳定性。 Objective: To investigate the effects of musk-frankincense compatibility treatment on the expression of tight junction-related proteins in prostatic epithelial cells in normal and chronic prostatitis rats. Methods: Eighty male Sprague-Dawley rats were randomly divided into 8 groups: normal control group, normal musk-frankincense group, normal musk group, normal frankincense group, model blank group, model musk-frankincense group, model musk group, model frankincense group . The rat model of chronic prostatitis was established. The rats in the model group were administered with a dose of 0.021 g / (kg · d) of musk and 1.05 g / (kg · d) of frankincense for 60 days respectively. The combined dose of the musk-frankincense group was blank Group given normal saline gavage. The rats in each group were given gavage for 3 days continuously, and the rats were sacrificed. Prostate tissues were harvested, and immunohistochemistry was used to detect the expression of tight junction protein 1, claudin 3, occludin, cytoplasm The expression of adhesion protein 1 (ZO-1). Results: In the pathological state, the expression of Claudin 1 increased only statistically significant. Musk and frankincense had different physiological and pathological effects on four proteins: musk (824.6 ± 393.3), frankincense (982.0 ± 334.0) or compatibility (1 088.1 ± 640.2) (P <0.05, P <0.01). For Claudin 3, the expression of Claudin was up-regulated (1009.5 ± 243.6, P <0.05) and the single musk (597.5 ± 80.7) The difference was not statistically significant. Musk (678.4 ± 255.1) could reduce the up-regulated expression of frankincense; the effect of single musk (693.0 ± 424.8), frankincense (732.1 ± 302.0) or compatibility treatment (560.2 ± 202.3) Statistical analysis showed that there was no significant difference in the expression of ZO-1 between musk (290.0 ± 166.8) and frankincense (419.7 ± 108.1) alone, and the expression of ZO-1 was significantly down-regulated after the combination treatment (197.7 ± 98.2, P <0.05). Under the pathological conditions of chronic prostatitis, the expressions of tight junction protein 1 (823.0 ± 100.1) and laminin (1 160.0 ± 32.2) in rat prostatic tissue were significantly increased (P <0.01, P <0.05) (468.4 ± 220.4) or compatibility (335.1 ± 204.0) with down-regulation of tight junction protein 1 expression (P <0.05); musk alone (700.1 ± 223.7) or frankincense (744.6 ± 94.5) (P <0.05). The expression of laminin could be down-regulated by single musk (749.6 ± 321.7), frankincense (615.0 ± 221.0) or compatibility (505.8 ± 523.7) (213.5 ± 24.9), which was significantly lower than that of musk (P <0.05). CONCLUSION: Musk and frankincense have selective and bidirectional regulation of the proteins involved in the regulation of tight junctions in the epithelial barrier of the prostate under physiologic / pathological conditions, and mainly through the down-regulation of ZO-1 protein to promote permeability. By regulating the expression of tight junction protein 1, tight junction protein 3, blocking protein expression to maintain structural stability.
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