[關(guān)鍵詞]
[摘要]
目的 探討桂枝茯苓膠囊對(duì)大鼠良性前列腺增生(BPH)的作用機(jī)制。方法 72只雄性SPF級(jí)SD大鼠隨機(jī)分為6組:對(duì)照組、模型組、非那雄胺(陽(yáng)性藥,2 mg·kg-1)組和桂枝茯苓膠囊低、中、高劑量(0.87、1.74、3.47 g·kg-1)組,每組12只;除對(duì)照組外,其余5組sc雌、雄激素(0.05 mg·kg-1苯甲酸雌二醇和4 mg·kg-1丙酸睪酮),每天1次,連續(xù)28 d,建立BPH模型。同時(shí)ig給予藥物,每天給藥1次,連續(xù)給藥28 d。HE染色觀察前列腺組織病理變化;ELISA法檢測(cè)血清中雙氫睪酮(DHT)、雌二醇(E2)和前列腺中DHT、E2、轉(zhuǎn)化生長(zhǎng)因子β1(TGF-β1)、表皮生長(zhǎng)因子(EGF)的水平;Western blotting檢測(cè)前列腺中增殖細(xì)胞核抗原(PCNA)、雄激素受體(AR)、雌激素受體α(ERα)、胰島素樣生長(zhǎng)因子(IGF-1)、成纖維細(xì)胞生長(zhǎng)因子7(FGF-7)、缺氧誘導(dǎo)因子1α(HIF-1α)和血管內(nèi)皮生長(zhǎng)因子(VEGF)蛋白表達(dá)水平;實(shí)時(shí)熒光定量PCR(qRT-PCR)法檢測(cè)前列腺中PCNA、FGF-7、IGF-1和VEGF mRNA表達(dá)水平。結(jié)果 與對(duì)照組比較,模型組前列腺指數(shù)顯著增加(P<0.01),前列腺上皮厚度顯著增加(P<0.01),前列腺顯著增生,間質(zhì)充血、水腫;血清中DHT和前列腺中DHT、E2、EGF水平顯著升高(P<0.05、0.01);前列腺中TGF-β1水平顯著降低(P<0.05),PCNA、AR、ERα、FGF-7、IGF-1、HIF-1α和VEGF蛋白表達(dá)水平及PCNA、FGF-7、IGF-1和VEGF mRNA表達(dá)水平顯著升高(P<0.05、0.01)。與模型組比較,桂枝茯苓膠囊高劑量顯著降低前列腺指數(shù)(P<0.05),各劑量均顯著減小前列腺上皮厚度(P<0.01),減輕間質(zhì)充血、水腫;桂枝茯苓膠囊低劑量顯著升高前列腺中TGF-β1水平,顯著降低EGF水平(P<0.05)以及PCNA、FGF-7、VEGF mRNA的表達(dá)(P<0.01),顯著下調(diào)前列腺中AR蛋白表達(dá)水平(P<0.05);中劑量能顯著升高前列腺中TGF-β1水平,顯著降低EGF水平(P<0.05、0.01)以及PCNA、VEGF mRNA的表達(dá)水平(P<0.01),顯著下調(diào)IGF-1蛋白表達(dá)水平(P<0.05);高劑量顯著升高前列腺中TGF-β1水平,顯著降低血清中DHT水平、前列腺中DHT、E2、EGF水平(P<0.05、0.01)以及PCNA、FGF-7、IGF-1和VEGF mRNA的表達(dá)水平(P<0.01),顯著下調(diào)PCNA、AR、ERα、FGF-7、IGF-1、HIF-1α和VEGF蛋白表達(dá)(P<0.05、0.01)。結(jié)論 桂枝茯苓膠囊對(duì)BPH有明顯的治療作用,其作用機(jī)制可能包括降低DHT、E2水平,抑制E2與ERα結(jié)合及AR信號(hào)通路;并且升高TGF-β1水平、降低EGF水平,抑制細(xì)胞增殖、誘導(dǎo)細(xì)胞凋亡,下調(diào)FGF-7、IGF-1、HIF-1α和VEGF生長(zhǎng)因子的表達(dá),抑制細(xì)胞增殖與血管生成。
[Key word]
[Abstract]
Objective To explore the mechanism of Guizhi Fuling capsule on benign prostatic hyperplasia (BPH) in rats.Methods 72 male SPF SD rats were randomly divided into six groups:control group, model group, finasteride (positive drug, 2 mg·kg-1) and Guizhi Fuling Capsules low-dose, medium-dose and high-dose (0.87, 1.74, 3.47 g·kg-1) groups, with 12 rats in each group. Except the control group, the other five groups were given male and female hormones (0.05 mg·kg-1 estradiol benzoate and 4 mg·kg-1 testosterone propionate), once a day for 28 d, and the BPH model was established. At the same time, the therapeutic drug was given by ig, with a volume of 10 mL·kg-1, once a day, for 28 d. The pathological changes of prostate tissue were observed by HE staining. The contents of DHT and E2 in serum, DHT, E2, TGF-β1 and EGF in prostate tissue were detected by ELISA. The protein expressions of PCNA, AR, ERα, IGF-1, FGF-7, HIF-1α and VEGF in prostate were detected by Western blotting. The mRNA expression of PCNA, FGF-7, IGF-1 and VEGF in prostate was detected by qRT-PCR. Results Compared with control group, the prostate index and epithelial thickness of prostate in the model group were significantly increased (P < 0.01), the prostatic hyperplasia, interstitial congestion and edema were significantly increased, and the contents of DHT in serum and DHT, E2 and EGF in prostate were significantly increased (P < 0.05 and 0.01). The content of TGF-β1 in prostate was significantly decreased (P < 0.05), and the protein expression levels of PCNA, AR, ERα, FGF-7, IGF-1, HIF-1α and VEGF and the mRNA expression levels of PCNA, FGF-7, IGF-1 and VEGF in prostate were significantly increased (P < 0.05 and 0.01). Compared with the model group, the high-dose group of Guizhi Fuling Capsule significantly reduced the prostate index (P < 0.05), while the low-dose, medium, and highdose groups significantly reduced the thickness of the prostate epithelium (P < 0.01), reducing prostate hyperplasia, interstitial congestion, and edema. The low-dose group of Guizhi Fuling Capsule significantly increased TGF-β1 content in the prostate gland, significantly reducing EGF content (P < 0.05) and the expression of PCNA, FGF-7, and VEGF mRNA (P < 0.01), significantly downregulating the expression level of AR protein in the prostate (P < 0.05). The medium dose group of Guizhi Fuling Capsule can significantly increase TGF-β1 content in the prostate gland, significantly reducing EGF content (P < 0.05 and 0.01) and the expression levels of PCNA and VEGF mRNA (P < 0.01), significantly downregulating the expression level of IGF-1 protein (P < 0.05). The high-dose group of Guizhi Fuling Capsule significantly increased TGF-β1 content in the prostate gland, significantly reducing the levels of DHT in serum, DHT, E2, EGF in prostate (P < 0.05 and 0.01), as well as the expression levels of PCNA, FGF-7, IGF-1, and VEGF mRNA (P < 0.01), significantly downregulating PCNA, AR, ERα、FGF-7, IGF-1, HIF-1α and VEGF protein expression (P < 0.05 and 0.01). Conclusion Guizhi Fuling capsule has obvious therapeutic effect on benign prostatic hyperplasia, which may reduce DHT and E2 content, inhibit E2 and ERα binding and AR signaling pathway. Moreover, it increased the content of TGF-β1 and decreased the content of EGF, inhibited cell proliferation and induced cell apoptosis, down-regulated the expression of FGF-7, IGF-1, HIF-1α and VEGF growth factors, and inhibited cell proliferation and was related to angiogenesis.
[中圖分類(lèi)號(hào)]
R285.5
[基金項(xiàng)目]
2021年國(guó)家中醫(yī)藥管理局岐黃學(xué)者項(xiàng)目(國(guó)中醫(yī)藥人教函[2022]6號(hào))