目的 觀察五子衍宗丸對(duì)高脂飲食誘導(dǎo)肥胖大鼠生精功能的影響，并從睪丸內(nèi)質(zhì)網(wǎng)應(yīng)激角度探討可能機(jī)制。方法 30 只雄性 SD 大鼠隨機(jī)分為對(duì)照組、模型組和五子衍宗丸低、中、高劑量（0.5、1.0、2.0 g·kg^-1）組。對(duì)照組給予普通飲食，其余組給予高脂飼料喂養(yǎng)，16 周后各組 ig 給予相應(yīng)劑量藥物，連續(xù) 4 周；取附睪進(jìn)行精子功能分析；血清 ELISA 分析性激素水平變化；蘇木素-伊紅染色觀察睪丸組織病理改變；透射電鏡觀察睪丸支持細(xì)胞線粒體和內(nèi)質(zhì)網(wǎng)結(jié)構(gòu)改變；免疫熒光染色檢測(cè)睪丸線粒體融合蛋白 2（Mfn2）及內(nèi)質(zhì)網(wǎng)應(yīng)激標(biāo)識(shí)蛋白葡萄糖調(diào)節(jié)蛋白 78（GRP78）的定位及表達(dá)；Western blotting 檢測(cè)睪丸組織 Mfn2 和蛋白激酶 R 樣內(nèi)質(zhì)網(wǎng)激酶（PERK）通路 相 關(guān) 蛋 白 相 對(duì) 表 達(dá) 。結(jié)果 與對(duì)照組比較， 模型組大鼠體質(zhì)量明顯增加（P＜0.01），血清雌二醇水平明顯升高（P＜0.01），睪酮水平明顯降低（P＜0.01）；睪丸生精細(xì)胞排列稀疏，細(xì)胞變性；精子活力明顯降低（P＜0.05），精子畸形率明顯升高（P＜0.01）；睪丸支持細(xì)胞內(nèi)質(zhì)網(wǎng)和線粒體腫脹，呈空泡狀，部分網(wǎng)膜斷裂，Mfn2 表達(dá)明顯降低，GRP78 熒光表達(dá)明顯增加；PERK、 活 化 轉(zhuǎn) 錄 因 子 -4 （ATF4） 和 CHOP 蛋白量明顯升高（P＜0.01）。與模型組比較，五子衍宗丸高劑量組體質(zhì)量減輕明顯（P＜0.05）；中、高劑量組血清雌二醇水平明顯降低（P＜0.05、0.01），睪酮水平明顯升高（P＜0.05、0.01）；高劑量組精子活力明顯升高（P＜0.05），精子畸形率明顯降低（P＜0.05）；睪丸病變減輕，排列較致密，支持細(xì)胞內(nèi)質(zhì)網(wǎng)和線粒體腫脹改善；Mfn2 熒光表達(dá)逐漸增強(qiáng)，蛋白相對(duì)表達(dá)明顯增加 （P＜0.05、0.01），GRP78 熒光表達(dá)逐漸減弱；低劑量組 ATF4 蛋白量明顯降低 （P＜0.01），中劑量組PERK、ATF4 蛋白量明顯降低（P＜0.05、0.01），高劑量組 PERK、ATF4 和 CHOP 蛋白量均明顯降低（P＜0.01）。結(jié)論 五子衍宗丸可改善高脂飲食誘導(dǎo)肥胖引起的大鼠生精障礙，其機(jī)制可能與調(diào)控睪丸支持細(xì)胞Mfn2表達(dá)，減輕內(nèi)質(zhì)網(wǎng)應(yīng)激有關(guān)。

Objective To observe the effect of Wuzi Yanzong Pill (WYP) on spermatogenic function in obese rats induced by high fat diet, and to explore the possible mechanism from ERS in testis. Methods Totally 30 male SD rats were divided into control group, model group, low, middle and high dose (0.5, 1.0, and 2.0 g·kg^-1) groups of WYP randomly. The control group was fed with normal diet, the rest were fed with high-fat diet. After 16 weeks, the corresponding doses of drugs in each group were gavaged for four weeks continuously. the rats were dissected, and sperms in comepididymis were collected for the function analysis. Serum ELISA was used to analyze the changes of sex hormone levels. The pathological changes of testis were observed by hematoxylin-eosin staining. Transmission electron microscopy (TEM) was used to observe the structural changes of the mitochondria and endoplasmic reticulum in sertoli cells. The localization and expression of Mfn2 and GRP78 in testis were detected by immunofluorescence staining. The expression of Mfn2 and PERK pathway related proteins in testis were detected by Western blotting. Results Compared with control group, the model group rats showed a significant increase in body weight (P<0.01), a marked increase in serum estradiol level (P<0.01), and a marked decrease in serum testosterone level (P<0.01); the arrangement of spermatogenic cells in the testes was sparse, and the cells were degenerated; the sperm motility was significantly lower (P<0.05), and the sperm malformation rate was markedly increased (P<0.01); the rough endoplasmic reticulum and mitochondria in the Leydig cells were swollen and hollow, some of the network was broken, the expression of Mfn2 was markedly reduced, and the GRP78 fluorescence expression was markedly increased; the protein levels of PERK, activated transcription factor-4 (ATF4), and CHOP were markedly increased (P<0.01). Compared with the model group, the high-dose group of Wuzi Yanzong Pill showed a significant reduction in body weight (P<0.05); the serum estradiol level was markedly lower in the middle and high-dose groups (P<0.05, 0.01), and the serum testosterone level was markedly higher (P<0.05, 0.01); the sperm motility was significantly improved in the high-dose group (P<0.05), and the sperm malformation rate was markedly reduced (P<0.05); the pathological changes in the testes were alleviated, the arrangement was denser, and the rough endoplasmic reticulum and mitochondria in the Leydig cells were improved; the fluorescence expression of Mfn2 gradually increased, and the relative protein expression was markedly increased (P<0.05, 0.01), while the GRP78 fluorescence expression gradually decreased; the ATF4 protein level was markedly reduced in the low-dose group (P<0.01), the PERK and ATF4 protein levels were markedly reduced in the middle-dose group (P<0.05, 0.01), and the PERK, ATF4, and CHOP protein levels were all markedly. Conclusion Wuzi Yanzong Pill can ameliorate spermatogenesis disorder induced by high fat diet in rats, which may be related to the regulation the expression of Mfn2 and alleviate the endoplasmic reticulum stress in sertoli cells.

R285.5

天津市衛(wèi)生健康科技項(xiàng)目（TJWJ2022MS048）