[Mechanism of Buyang Huanwu Decoction glycosides against atherosclerotic inflammation through NF-κB signaling pathway].

This study aims to explore the effect of Buyang Huanwu Decoction glycosides on the inflammatory response of apolipoprotein E~(-/-)(ApoE~(-/-)) mice and RAW264.7 cells through nuclear factor kappa-B(NF-κB) signaling pathway. In the in vivo experiment, ApoE~(-/-) mice were fed with high-fat diets for 12 weeks to induce the animal model of atherosclerosis, and 75 µg·mL~(-1) oxidized low-density lipoprotein(Ox-LDL) incubated RAW264.7 cells for 24 h to establish the atherosclerosis cell model. Automatic biochemical analyzer, hematoxylin-eosin(HE) staining, enzyme-linked immunosorbent assay(ELISA), Western blot, and droplet digital polymerase chain reaction(PCR) were used to determine the blood lipid levels, aortic intimal thickness, inflammatory factor content, NF-κB pathway-related proteins, and mRNA expression levels, and evaluate arterial atherosclerotic lesions and anti-atherosclerotic mechanisms of the drug. The model of atherosclerosis was successfully established in ApoE~(-/-) mice after 12 weeks of feeding with high-fat diets. In the model group, the plasma levels of total cholesterol(TC), triglyceride(TG), and low-density lipoprotein cholesterol(LDL-C) were increased(P<0.01), the intima of the blood vessels was thickened, the levels of inflammatory factors tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were increased, and the protein and mRNA expressions of NF-κB and inhibitor of NF-κB(IκBα) were significantly increased as compared with the control group. Compared with the model group, the high-dose Buyang Huanwu Decoction glycoside group decreased the plasma levels of TC, TG, and LDL-C, reduced the plaque area and thickness and the content of inflammatory factor TNF-α, and inhibited the protein and mRNA expressions of NF-κB and IκBα, with the effect same as Buyang Huanwu Decoction. In the in vivo experiment, 75 µg·mL~(-1) Ox-LDL stimulated RAW264.7 cells for 24 h to successfully establish a foam cell model. As compared with the control group, the nuclear amount of NF-κB and the protein and mRNA expressions of IκBα in the model group increased. Compared with the model group, the middle-dose and high-dose Buyang Huanwu Decoction glycoside groups decreased the nuclear amount of NF-κB and the protein and mRNA expressions of IκBα. The above results show that the glycosides are the main effective substances of Buyang Huanwu Decoction against atherosclerosis, which inhibit the NF-κB pathway and reduce the inflammatory response, thus playing the role against atherosclerotic inflammation same as Buyang Huanwu Decoction.

[Effect of Polygonati Rhizoma in improving pyroptosis injury of diabetic macroangiopathy via NLRP3/caspase-1/GSDMD pathway].

This study aims to explore the influence of Polygonati Rhizoma on the pyroptosis in the rat model of diabetic macroangiopathy via the NOD-like receptor thermal protein domain associated protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/gasdermin D(GSDMD) pathway. The rat model of diabetes was established by intraperitoneal injection of streptozotocin(STZ) combined with a high-fat, high-sugar diet. The blood glucose meter, fully automated biochemical analyzer, hematoxylin-eosin(HE) staining, enzyme-linked immunosorbent assay, immunofluorescence, immunohistochemistry, and Western blot were employed to measure blood glucose levels, lipid levels, vascular thickness, inflammatory cytokine levels, and expression levels of pyroptosis-related proteins. The mechanism of pharmacological interventions against the injury in the context of diabetes was thus explored. The results demonstrated the successful establishment of the model of diabetes. Compared with the control group, the model group showed elevated levels of fasting blood glucose, total cholesterol(TC), triglycerides(TG) and low-density lipoprotein cholesterol(LDL-c), lowered level of high-density lipoprotein cholesterol(HDL-c), thickened vascular intima, and elevated serum and aorta levels of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß) and interleukin-18(IL-18). Moreover, the model group showed increased NLRP3 inflammasomes and up-regulated levels of caspase-1 and GSDMD in aortic vascular cells. Polygonati Rhizoma intervention reduced blood glucose and lipid levels, inhibited vascular thickening, lowered the levels of TNF-α, IL-1ß, IL-18 in the serum and aorta, attenuated NLRP3 inflammasome expression, and down-regulated the expression levels of caspase-1 and GSDMD, compared with the model group. In summary, Polygonati Rhizoma can slow down the progression of diabetic macroangiopathy by inhibiting pyroptosis and alleviating local vascular inflammation.

[Efficacy of Zhibai Dihuang Decoction on energy metabolism of spermatogenic cells in Ureaplasma urealyticum-infected rats: A study based on AMPK and PPARα signaling pathways].

OBJECTIVE: To study the effect of the serum containing Zhibai Dihuang Decoction (ZDD) on the energy metabolism of spermatogenic cells infected with Ureaplasma urealyticum (UU) in rats and its action mechanism. METHODS: Healthy male SD rats were randomly divided into six groups, normal control, UU-infection (UUI) model control, doxycycline, and low-, medium- and high-dose ZDD-containing serum. After successful establishment of the UUI model in vivo in the latter five groups, the rats in the normal control group were treated with simple serum and those in the latter five with respective agents. Then primary spermatogenic cells were harvested from the rats for examination of the apoptosis of spermatogenic cells, contents of lactate dehydrogenase (LDH) and adenosine triphosphate (ATP), glucose disposal rate (GDR) and expressions of AMPK and PARα proteins in the spermatogenic cells, and other related parameters. RESULTS: The apoptosis rate of the spermatogenic cells was dramatically increased in the UUI model controls compared with that in the normal controls (ï¼»49.24 ± 0.86ï¼½% vs ï¼»10.09 ± 0.52ï¼½%, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»11.21 ± 1.02ï¼½%, ï¼»30.64 ± 0.99ï¼½%, ï¼»35.54 ± 1.17ï¼½% and ï¼»42.95 ± 1.31ï¼½%) in comparison with that in the UUI model control group (P < 0.01).The content of LDH in the spermatogenic cells was also remarkably increased in the UUI model controls compared with that in the normal controls (ï¼»201.12 ± 2.88ï¼½ vs ï¼»60.72 ± 1.83ï¼½) mU/ml, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»90.66 ± 1.61ï¼½, ï¼»94.74 ± 1.20ï¼½, ï¼»101.24 ± 2.03ï¼½ and ï¼»111.04 ± 3.35ï¼½ mU/ml) in comparison with that in the UUI model control group (P < 0.01). The GDR in the spermatogenic cells was markedly reduced in the UUI model controls compared with that in the normal controls (ï¼»49.42 ± 1.70ï¼½% vs ï¼»99.86 ± 1.26ï¼½%, P < 0.01), but significantly elevated in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»86.90 ± 2.03ï¼½%, ï¼»84.14 ± 1.21ï¼½%, ï¼»80.30 ± 1.37ï¼½% and ï¼»75.18 ± 1.76ï¼½% in comparison with that in the UUI model control group (P < 0.01). The content of ATP was also dramatically decreased in the UUI model controls compared with that in the normal controls (ï¼»19.76 ± 1.46ï¼½ vs ï¼»58.94 ± 1.95ï¼½ µmol/L, P < 0.01), but significantly increased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»48.34 ± 1.34ï¼½, ï¼»42.82 ± 1.30ï¼½, ï¼»38.70 ± 2.03ï¼½ and ï¼»34.78 ± 0.82ï¼½ µmol/L) in comparison with that in the UUI model control group (P < 0.01). The mitochondrial membrane potential was remarkably elevated in the UUI model controls compared with that in the normal controls (ï¼»8.53 ± 0.71ï¼½% vs ï¼»2.43 ± 0.25ï¼½%, P < 0.01), but markedly reduced in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»3.92 ± 0.36ï¼½%, ï¼»4.43 ± 0.27ï¼½%, ï¼»4.65 ± 0.22ï¼½% and ï¼»4.88 ± 0.10ï¼½% in comparison with that in the UUI model control group (P < 0.01). The phosphorylation levels of AMPK and PPARα proteins were significantly up-regulated in the UUI model controls compared with that in the normal controls (P < 0.01), but down-regulated in a dose-dependent manner in the ZDD groups. CONCLUSIONS: Zhibai Dihuang Decoction can significantly improve the damage to the mitochondrial structure and inhibit UU infection-induced apoptosis of spermatogenic cells and secretion of LDH by increasing the ATP content and GDR and regulating the phosphorylation of AMPK and PARα signaling pathway.

[Xiongcan Yishen Prescription upregulates the expressions of eNOS and cGMP in the penile tissue of ED rats with liver depression and kidney deficiency].

OBJECTIVE: To investigate the effect of Xiongcan Yishen Prescription (XYP) on the expressions of eNOS and cGMP in the penile tissue of ED rats with liver depression and kidney deficiency (LDKD). METHODS: The model of ED-LDKD was established in 30 eight-week-old SPF-class male SD rats by injecting hydrocortisone intramuscularly and binding the limbs for 14 days, and another 10 rats were taken as blank controls. Then, the model rats were randomized into six groups of equal number and treated intragastrically with distilled water (model control), tadalafil tablets at 0.52 mg/kg/d (tadalafil control), Shugan Yiyang Capsules 0.3125 g/kg/d (SYC control), and XYP at 10.4 g/kg/d (low-dose XYP), 20.8 g/kg/d (medium-dose XYP) and 41.6 g/kg/d (high-dose XYP), bid, for 28 successive days, respectively. Before and after modeling and after 28-day treatment, the animals were subjected to tail suspension and mating tests. The next day after medication, the penile tissues of the rats were harvested for determining the expression levels of eNOS and cGMP proteins by immunohistochemical analysis of the mean optical density. RESULTS: Compared with the model controls, the rats of the high-, medium- and low-dose XYP and SYC control groups all showed significant decreases in the tail suspension time (ï¼»3.17 ± 0.11ï¼½ vs ï¼»2.58 ± 0.25ï¼½, ï¼»2.52 ± 0.31ï¼½, ï¼»2.51 ± 0.3ï¼½ and ï¼»2.57 ± 0.29ï¼½ min, P < 0.05) and mount latency (ML) (ï¼»9.23 ± 0.11ï¼½ vs ï¼»1.21 ± 0.12ï¼½, ï¼»2.17 ± 0.16ï¼½, ï¼»2.26 ± 0.13ï¼½, ï¼»1.23 ± 0.15ï¼½ and ï¼»2.48 ± 0.18ï¼½ min, P < 0.05) but increases in mount frequency (MF) (ï¼»0.48 ± 0.18ï¼½ vs ï¼»3.29 ± 0.11ï¼½, ï¼»3.18 ± 0.11ï¼½, ï¼»3.05 ± 0.05ï¼½, ï¼»3.23 ± 0.12ï¼½ and ï¼»3.2 ± 0.28ï¼½ times, P < 0.05) and intromission frequency (IF) (ï¼»0.8 ± 0.84ï¼½ vs ï¼»11.8 ± 0.84ï¼½, ï¼»11.2 ± 1.48ï¼½, ï¼»9.4 ± 1.14ï¼½, ï¼»11.4 ± 1.14ï¼½ and ï¼»10 ± 1.22ï¼½ times, P < 0.05). The eNOS and cGMP proteins were mainly expressed in the nucleus and cytoplasm of the arterial and venous endothelial cells and sinusoidal endothelial cells of the cavernous, as brownish yellow particles in a scattered and focal pattern. Both the expressions of eNOS and cGMP in the penile tissue were remarkably upregulated in the high-, medium- and low-dose XYP and SYC control groups as compared with those in the model control (P < 0.05) but exhibited no statistically significant difference between the tadalafil and model control groups (P > 0.05). CONCLUSIONS: Xiongcan Yishen Prescription can relieve the depression symptoms, increase the mount frequency, activate the NO/cGMP pathway, and upregulate the expressions of eNOS and cGMP in the penile tissue of ED rats with liver depression and kidney deficiency.

[Xiongcan Yishen Prescription up-regulates the expressions of cholesterol transport proteins, steroidogenic enzymes and SF-1 in the Leydig cells of rats with late-onset hypogonadism].

OBJECTIVE: To investigate the effects of Xiongcan Yishen Prescription (XYP) on the expressions of cholesterol transport proteins, steroidogenic enzymes and steroidogenic factor-1 (SF-1) in the Leydig cells of the rats with late-onset hypogonadism (LOH). METHODS: Twenty-five 18-month-old male SD rats were randomly divided into five groups of equal number, LOH model control, testosterone propionate (TP) and low-, medium- and high-dose XYP, and another 5 two-month-old male SD rats included as normal controls. After modeling, the animals in the TP group were treated by intramuscular injection of TP at 5.21 mg/kg qd alt, those in the low-, medium- and high-dose XYP groups intragastrically with XYP at 10.4, 20.8 and 41.6 g/kg qd alt respectively, and those in the LOH model and normal control groups with saline, all for 28 successive days. Then, all the rats were sacrificed for determination of the expressions of the cholesterol transport proteins StAR and TSPO, steroidogenic enzymes CYP11A1, HSD3B7 and HSD17B4, and SF-1 in the Leydig cells by Western blot. RESULTS: The expressions of StAR, TSPO, CYP11A1, HSD3B7, HSD17B4 and SF-1 in the Leydig cells were significantly decreased in the LOH model controls compared with those in the normal controls (P< 0.05), but remarkably increased in the low-, medium- and high-dose XYP groups in comparison with those in the LOH model control group (P< 0.05). CONCLUSIONS: Xiongcan Yishen Prescription can up-regulate the expressions of the cholesterol transport proteins StAR and TSPO, steroidogenic enzymes CYP11A1, HSD3B7 and HSD17B4, and SF-1 in the rat Leydig cells, which might be one of the possible mechanisms of the prescription in the treatment of LOH.

[Impact of Ureaplasma urealyticum infection on the MRPS22 protein expression in rat spermatogenic cells and intervening effect of Zhibai Dihuang Decoction].

OBJECTIVE: To investigate the impact of Ureaplasma urealyticum (UU) infection (UUI) on the expression of the mitochondrial ribosomal protein S22 (MRPS22) in rat spermatogenic cells and the intervening effect of Zhibai Dihuang Decoction (ZBDH). METHODS: Forty-five SD male rats were randomly divided into four groups of equal number: normal control, UUI model control, ZBDH and azithromycin, and the UUI model was made by bladder injection of the standard UU strain in the latter three groups. After modeling, the rats in the ZBDH and azithromycin groups were treated intragastrically with ZBDH at 1 g/kg/d and azithromycin at 0.105 g/kg/d respectively, while those in the normal and UUI model control groups with normal saline at 1 ml/kg/d. At 21 days after intervention, all the animals were sacrificed and their testes harvested for observation of the apoptosis and mitochondrial ultrastructure of the spermatogenic cells, measurement of the mitochondrial membrane potential (MMP) by flow cytometry, and determination of the mRNA and protein expressions of MRPS22 by RT-PCR and Western blot, respectively. RESULTS: The apoptotic index of the rat sperma-togenic cells was significantly higher in the UUI model control than in the ZBDH and azithromycin groups (ï¼»11.23 ± 1.65ï¼½ % vs ï¼»6.62 ± 0.49ï¼½ % and ï¼»7.82 ± 0.81ï¼½ %, P < 0.01), but lower in the ZBDH than in the azithromycin group (P < 0.05). The mitochondrial ultrastructure of the spermatogenic cells was markedly improved in the ZBDH and azithromycin groups as compared with that in the model control. The MMP level was remarkably lower in the model control than in the normal control (ï¼»8.77 ± 1.73ï¼½ % vs ï¼»22.33 ± 1.66ï¼½ %, P < 0.01), but higher in the ZBDH (ï¼»18.26 ± 1.32ï¼½ %) than in the model control (P < 0.01) and the azithromycin group (ï¼»15.91 ± 1.69ï¼½ %) (P < 0.01). The mRNA and protein expressions of MRPS22 were significantly lower in the model control (8.02 ± 3.21 and 22.65 ± 5.31) than in the normal control (15.43 ± 2.54 and 33.31 ± 7.09), ZBDH (11.26 ± 3.82 and 33.35 ± 3.96), and azithromycin group (8.79 ± 2.03 and 28.11 ± 4.13) (all P < 0.01), but both higher in the ZBDH than in the azithromycin group (P < 0.01). There was a positive correlation between the MRPS22 protein expression and MMP (r = 0.639, P < 0.01). CONCLUSIONS: UU infection induces the apoptosis of rat spermatogenic cells by inhibiting the mRNA and protein expressions of MRPS22 and reducing the mitochondrial membrane potential, while ZBDH can decrease the apoptosis of spermatogenic cells by improving the mRNA and protein expressions of MRPS22 and enhancing the mitochondrial membrane potential.

[Zhibai Dihuang Decoction improves sperm mitochondrial permeability transition in rats with ureaplasma urealyticum infection].

OBJECTIVE: To investigate the effects of Zhibai Dihuang Decoction (ZDD) on sperm mitochondrial permeability transition (MPT) in the rat model of ureaplasma urealyticum (UU) infection (UUI). METHODS: Ninety male SD rats were randomly divide into five groups: normal control, UUI model control, ZDD, doxycycline, and ZDD + doxycycline. The UUI model was established in the latter four groups of rats by UU injection into the bladder. On the second day after modeling, the animals of the normal control and UUI model control groups were treated intragastrically with 0.9% sodium chloride solution and those in the other groups with corresponding drugs, all for 21 consecutive days. At 24 hours after drug withdrawal, epididymal samples were obtained for detection of the protein and mRNA expressions of VDAC2 and ANT4 in the sperm mitochondria by RT-PCR and Western blot respectively and determination of the contents of adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) and energy charge (EC) in the sperm mitochondria by high-performance liquid chromatography. RESULTS: The protein expressions of VDAC2 and ANT4 in the rat sperm mitochondria were 0.626 ± 0.074 and 0.527 ± 0.096 in the normal control group, 0.039 ± 0.011 and 0.044 ± 0.011 in the UUI model control group, 0.101 ± 0.037 and 0.127 ± 0.040 in the ZDD group, 0.236 ± 0.070 and 0.253 ± 0.054 in the doxycycline group, and 0.475 ± 0.064 and 0.367 ± 0.086 in the ZDD + doxycycline group, significantly lower in the UUI model control than in the normal control group (P<0.05 and P<0.01), but remarkably higher in the doxycycline and ZDD + doxycycline groups than in the UUI model control (P<0.01) and the ZDD group (P<0.05 and P<0.01), and the expression of VDAC2 was markedly higher in the ZDD + doxycycline than in the doxycycline group (P<0.01). The mRNA expressions of VDAC2 and ANT4 were 0.008 ± 0.001 035 and 0.026 50 ± 0.003 401 in the normal control group, 0.000 79 ± 0.000 226 and 0.001 64 ± 0.000 205 in the UUI model controls, 0.002 06 ± 0.000 861 and 0.005 04 ± 0.002 537 in the ZDD group, 0.003 34 ± 0.000 229 and 0.008 57 ± 0. 000 690 in the doxycycline group, and 0.004 85 ± 0.000 495 and 0.013 13 ± 0.000 826 in the ZDD + doxycycline group, significantly lower in the UUI model control than in the normal control group (P<0.05 and P<0.01), but remarkably higher in the ZDD, doxycycline and ZDD + doxycycline groups than in the UUI model controls (P<0.01) as well as in the doxycycline and ZDD + doxycycline groups than in the ZDD group (P<0.01) and in the ZDD + doxycycline than in the doxycycline group (P<0.01). The levels of ATP, ADP, AMP and EC in the sperm mitochondria were (203.41 ± 13.16) mg/L, (129.87 ± 14.68) mg/L, (149.05 ± 5.65) mg/L and 0.56 ± 0.01 in the normal control group, (96.22 ± 12.55) mg/L, (99.87 ± 3.28) mg/L, (212.53 ± 19. 43) mg/L and 0.36 ± 0.03 in the UUI model control group, (101.99 ± 5.97) mg/L, (104.99 ± 16.40) mg/L, (183.97 ± 12.43) mg/L and 0.40 ± 0.01 in the ZDD group, (159.44 ± 33.16) mg/L, (118.51 ± 12.99) mg/L, (160.64 ± 14.19) mg/L and 0.50 ± 0.06 in the doxycycline group, and (194.07 ± 9.36) mg/L, (121.62 ± 9.41) mg/L, (150.21 ± 12.87) mg/L and 0.55 ± 0.01 in the ZDD + doxycycline group. The levels of ATP and EC were significantly lower and that of AMP higher in the UUI model control than in the normal control group (P<0.01), while the former two were remarkably higher and the latter one lower in the doxycycline and ZDD + doxycycline groups than in the UUI model controls (P<0.05 and P<0.01). Compared with the ZDD + doxycycline group, the ZDD group showed significantly decreased ATP and EC but increased AMP, while the doxycycline group exhibited decreases in both ATP and EC (P<0.05 and P<0.01). CONCLUSIONS: ZDD can upregulate the decreased protein and mRNA expressions of VDAC2 and ANT4 in the sperm mitochondria and improve sperm mitochondrial permeability transition and mitochondrial energy metabolism in rats with UU infection, which may be one of its action mechanisms in the treatment of UU infection-induced male infertility.

[Effect of Shirexiao waist hot-compress on the expressions of Th17/Treg-specific factors in the mouse model of experimental autoimmune prostatitis with damp heat syndrome].

OBJECTIVE: To investigate the effect of waist hot-compress with the Shirexiao (SRX) pad on the expressions of Th17/Treg-specific factors in the prostatic tissue of the mouse model of experimental autoimmune prostatitis (EAP) with damp heat syndrome, and explore its possible action mechanisms. METHODS: Twenty healthy male mice were included as normal controls and another 100 chosen for establishing the model of EAP with damp heat syndrome by subcutaneous injection of purified prostate protein solution from the Wistar rat and Freund's complete adjuvant using the TCM method. The model mice were randomly divided into five groups: model control, matrix, and low-, medium- and high-dose SRX. After chemical removal of the hair at lumbar vertebrae 1－3, the animals of the low-, medium- and high-dose SRX groups were treated with the SRX pad heated to 45℃ and externally applied to the non-hair area, qd, bid, and tid, respectively, 10 minutes each time, those of the matrix group with the vaseline pad, and those of the normal and model control groups with the saline pad. After 4 weeks of continuous treatment, all the mice were sacrificed for determination of the protein and mRNA expressions of RORγt and Foxp3 in the prostate tissue by Western blot and quantitative real-time PCR. RESULTS: The symptoms, signs and pathological changes of the EAP model mice were similar to the manifestations of chronic prostatitis. After intervention, the protein and mRNA expressions of Foxp3 were significantly down-regulated while those of RORγt markedly up-regulated in the EAP model group as compared with the normal control (P <0.05). In comparison with the model controls, the protein and mRNA expressions of RORγt were remarkably decreased in the medium- and high-dose SRX groups (P <0.05), that of the Foxp3 protein was markedly increased in the high-dose group (P <0.05), while that of Foxp3 mRNA exhibited no statistically significant difference in the low-, medium- or high-dose groups (P >0.05). CONCLUSIONS: The Shirexiao waist hot-compress therapy plays a positive role in the treatment of autoimmune prostatitis with damp heat syndrome by reducing the expression of RORγt, inhibiting the differentiation of Th17 and thus checking the differentiation imbalance of Th17/Treg.

[Effects of Zhibai Dihuang Decoction on mitochondrial cytochrome oxidase in the spermatogenic cells of rats with ureaplasma urealyticum infection].

OBJECTIVE: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (ï¼»1.03 ± 0.09ï¼½ vs ï¼»0.07 ± 0.03ï¼½ %, P<0.01), grade b sperm (ï¼»2.07 ± 0.52ï¼½ vs ï¼»0.35 ± 0.13ï¼½ %, P<0.01), straight line velocity (VSL) (ï¼»10.95 ± 0.98ï¼½ vs ï¼»6.78 ± 1.05ï¼½ µm/s, P<0.01), curvilinear velocity (VCL) (ï¼»42.03 ± 1.35ï¼½ vs ï¼»38.10 ± 7.65ï¼½ µm/s, P>0.05), average path velocity (VAP) (ï¼»16.22 ± 1.52ï¼½ vs ï¼»10.05 ± 1.80ï¼½ µm/s, P<0.01), and the mRNA expressions of COX Ⅰ (ï¼»2.25 ± 0.24ï¼½ vs ï¼»0.93 ± 0.10ï¼½ %, P<0.01) and Ⅱ (ï¼»6.72 ± 0.37ï¼½ vs ï¼»2.95 ± 0.78ï¼½ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ï¼»1.11 ± 0.30ï¼½ and ï¼»0.60 ± 0.19ï¼½%; grade b sperm: ï¼»2.40 ± 0.59ï¼½ and ï¼»1.32 ± 0.27ï¼½ %; VSL: ï¼»12.11 ± 1.62ï¼½ and ï¼»11.47 ± 1.21ï¼½ µm/s; VCL: ï¼»54.30 ± 2.35ï¼½ and ï¼»45.75 ± 1.64ï¼½ µm/s; VAP ï¼»18.40 ± 1.27ï¼½ and ï¼»16.69 ± 1.02ï¼½ µm/s; expression of COXⅠ mRNA: ï¼»1.86 ± 0.30ï¼½ and ï¼»1.74 ± 0.17ï¼½ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS: UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.

[Traditional Chinese Medicine promotes the development of andrology].

Andrology is an ancient branch of science which has gained a new development in the recent years and therefore has both traditional and modern characteristics. On the one hand, andrology keeps benefiting from the achievements of modern medicine and, on the other hand, it relies on the support of the theories of Traditional Chinese Medicine (TCM). An integration of Chinese and Western medical principles may further facilitate the development of andrology. TCM may contribute to the development of andrology by giving full play to its advantage as a psychosomatic medicine, enriching treatment strategies for male diseases with comprehensive TCM therapies, integrating the advantages of Western medicine to improve clinical efficacy, and normalizing the use of patent TCM drugs.

[Antioxidating and energy metabolism improving effects of Qiangjing Decoction on oligospermia and asthenospermia: An experimental study].

OBJECTIVE: To explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS). METHODS: We randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR. RESULTS: The concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05). CONCLUSION: Ornidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

[Zhibai Dihuang Decoction improves the activity of sperm mitochondrial respiratory chain complex in rats with Ureaplasma urealyticum infection].

OBJECTIVE: To investigate the effect of Zhibai Dihuang Decoction (ZDD) on the sperm mitochondrial respiratory chain complex (MRCC) in rats with Ureaplasma urealyticum (UU) infection. METHODS: Ninety male SD rats were randomly divided into five groups, sham operation, UU infection model control, ZDD (crude drug at 8.56 g per kg of the body weight per day), doxycycline (DC, at 20 mg per kg of the body weight per day), and ZDD+DC. The model of UU infection was established by injecting UU into the bladder of all the rats except those of the sham operation group. After modeling, the rats were treated intragastrically with respective drugs for 21 days and then executed and their epididymides harvested for examination of sperm quality and determination of the activities of sperm MRCCs I, II, III and IV by spectrophotometry. RESULTS: At 10 days after modeling, the UU-positive rates in the model control, sham operation, ZDD, DC and ZDD+DC groups were 92.9%, 0%, 33.3%, 26.7% and 20.0%, respectively, significantly higher in the model control than in the other groups (P<0.05). The epididymal sperm concentrations in the five groups were (0.97±0.23), (3.02±0.52), (1.21±0.35), (1.02±0.31) and (1.52±0.28) ×106 ml, the sperm motilities were (58.62±15.36), (80.45±7.21), (75.52±8.78), (68.43±10.25) and (78.25±7.67)%, and rates of grade a+b sperm were (6.15±1.02), (10.32±1.14), (10.12±1.08), (9.01+1.27) and (10.74±1.03)%, respectively, all remarkably lower in the model control than in the sham operation group (P<0.01), but markedly higher in the ZDD and ZDD+DC groups than in the model controls (P<0.05). The activities of MRCC I in the model control, sham operation, ZDD, DC and ZDD+DC groups were (31.54±16.25), (136.86±6.34), (100.68±14.41), (81.68±6.78) and (124.06±5.54) µmol/(min·mg), those of MRCC II were (9.50±3.86), (20.34±0.37), (10.88±1.04), (12.93±1.07) and (16.23±0.60) µmol/(min·mg), those of MRCC III were (5.58±1.79), (19.60±0.61), (11.34±1.35), (13.87±1.23) and (15.96±0.69) µmol/(min·mg), and those of MRCC IV were (9.54±1.34), (28.98±3.33), (17.02±2.04), (18.41±2.67) and (21.66±2.93) µmol/(min·mg), respectively, all significantly lower in the model control than in the sham operation group (P<0.01), with the activities of MRCCs I, III and IV remarkably higher in the ZDD, DC and ZDD+DC groups (P<0.01) and that of MRCC II higher in the DC and ZDD+DC groups than in the model control (P<0.05). CONCLUSIONS: ZDD can improve the epididymal sperm quality and the activity of the sperm MRCC in UU-infected rats, which may be one of the mechanisms of ZDD acting on male infertility caused by UU infection.

[Effect of Zhibai Dihuang Decoction on Expressions of TRPV1 and TRPV5 in Spermatogenic Cells of UU-infected Rats].

OBJECTIVE: To observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD. METHODS: Totally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot. RESULTS: The UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01). CONCLUSION: ZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

[Hepatocyte growth factor and male reproduction].

The hepatocyte growth factor ( HGF) is a multifunctional growth factor, which produces multiple biological effects by binding to the c-Met acceptor. This article reviews the biological properties of HGF, particularly those correlated with male reproduction, including its abilities to promote testis embryonic development, spermatogenesis, and testosterone synthesis of Leydig cells. HGF may provide a new insight into the treatment of male hypogonadism and infertility.

[Effect of Zhibai Dihuang Pill on uPA of Uu-infection infertility patients].

OBJECTIVE: To study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients. METHODS: Recruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed. RESULTS: The sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05). CONCLUSIONS: Infection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

[Effects of zhibal dihuang decoction on the pathological changes and the ultrastructure of the testicular tissue in the ureaplasma urealyticum-infected rats].

OBJECTIVE: To study the effects of Zhibai Dihuang Decocion (ZDD) on the pathological changes and the ultrastructure of the testicular tissue in the ureaplasma urealyticum (UU)-infected rats. METHODS: The UU infected animal models were established by the bladder inoculation. The 45 UU infected SD rats were randomly divided into four groups, i.e., the ZDD treatment group (at the daily dose of 2 g/100 g), the Minocycline group (at the daily dose of 10 mg/100 g), the model group, 15 in each group. Besides, another 15 rats were recruited as the sham-operation group. The medication was started 10 days after vaccination. Equal volume of normal saline was given to rats in the model group and the sham-operation group by gastrogavage for 22 successive days. Rats were sacrificed on the 2nd day of medication discontinuation. The testicle mass index was detected. The ultra-structure and the pathological changes of the testicular tissue were observed by optical microscope and transmission electron microscope. RESULTS: There was no significant difference in the rat testicular mass index (P>0.05). UU infection can lead to the pathological changes such as atrophy of seminiferous tubules, germ cell loss, and reduction of sperm cells in lumen, and to the ultrastructural changes such as spermatogenic cell nuclear membrane shrinkage, nuclear breakdown, and obvious edema of mitochondria. The pathological changes and the ultrastructures were improved in the medication groups. Rm and Rs the were not overlapping, and the difference was statistically significant (P<0.05). Rm, Rzh, and Rx were not overlapping, and the difference was statistically significant (P<0.05). Rzh and Rx were overlapping in 95% Cl with no statistical difference (P>0.05). CONCLUSIONS: UU infection can cause the pathological changes and the ultrastructural changes of the testicular tissue at the organic level and the cellular level. ZDD played therapeutic effects through ameliorating its pathological changes and the ultrastructural changes of spermatogenic cells.

[Musk and carterii birdw enhance the effect of polygonum extract on chronic non-bacterial prostatitis: an animal experimental study].

OBJECTIVE: To observe the effects of musk and carterii birdw on the pathology and the expressions of inflammatory cytokines in chronic non-bacterial prostatitis (CNBP) rats treated with polygonum extract. METHODS: Five male Wistar rats were used for the preparation of SC purified prostate protein solution, and another 48 randomly divided into four groups: polygonum extract, polygonum extract + musk and carterii birdw, CNBP model control and normal control. CNBP models were established by injecting SC purified prostate protein solution and Freund's complete adjuvant. At 60 days after modeling, the CNBP model control and normal control rats were treated with normal saline, and the other two groups with polygonum extract and polygonum extract + musk and carterii birdw, respectively (polygonum 1.575 g per kg per d, musk 0.021 g per kg per d, and carterii birdw 1.05 g per kg per d). After 14 days of continuous intragastric medication, all the rats were sacrificed for pathological examination, determination of the levels of TNF-alpha, IL-1beta, IL-6 and IL-8 in the prostate tissue homogenate by ELISA, and detection of the mRNA and protein expressions of inflammatory cytokines MCP-1 (CCL2) and CCR2 by RT-PCR and Western blot. RESULTS: The polygonum extract + musk and carterii birdw group showed apparent improvement in the structure of the prostate tissue but no inflammatory infiltration, as was quite obvious in the polygonum extract group. Polygonum extract + musk and carterii birdw significantly decreased the inflammatory cytokines TNF-alpha ( [11.04 +/- 4.07] pg/ml), IL-1beta ([16.94 +/- 4.26] pg/ml), IL-6 ([110.08 +/- 28.42] pg/ml) and IL-8 ([26.28 +/- 7.36] pg/ml) in the prostate tissue, as compared with polygonum extract alone ([63.21 +/- 21.37] pg/ml, [41.32 +/- 14.62] pg/ml, [177.64 +/- 42.65] pg/ml and [96.37 +/- 37.61] pg/ml) (P < 0.05, P < 0.01). The former also exhibited significantly lower expressions of MCP-1 mRNA (0.32 +/- 0.17), MCP-1 protein (0.28 +/- 0.15), CCR2 mRNA (0.28 +/- 0.11) and CCR2 protein (0.11 +/- 0.04) than either the model control group (1.15 +/- 0.39, 0.93 +/- 0.34, 0.83 +/- 0.26 and 0.93 +/- 0.34) (P < 0.01), or the polygonum extract group (0.65 +/- 0.27, 0.56 +/- 0.22, 0.78 +/- 0.24 and 0.25 +/- 0.09) (P < 0.05, P < 0.01). CONCLUSION: Musk and carterii birdw can enhance the effect of polygonum extract on chronic prostatitis, reduce inflammatory response and improve tissue repair of the prostate in rats.

[Effect of tianoan zhuangyang powder on the stAR protein expression of Leydig cells in model rats with partial androgen deficiency of aging male].

OBJECTIVE: To observe the effect of Tiancan Zhuangyang Powder (TCZYP) on the steroidogenic acute regulatory protein (stAR protein) expression of Leydig cells in model rats with partial androgen deficiency of aging male (PADAM). METHODS: PADAM rat model was prepared by cyclophosphamide induced reproductive system damage. Rats were randomly divided into four groups, i. e. the normal control group, the model group, the testosterone propionate group, and the TCZYP group. The general condition, body weight, tail suspension experiment and exhaustion swimming test, and the testicular index, etc. were observed to assess the state of rats. The serum total testosterone (TT), and free testosterone (FT) levels were detected by radioimmunoassay before and after treatment. The stAR protein expression level of Leydig cells were determined using immunohistochemical assay. Statistic analyses were performed. RESULTS: By modeling with cyclophosphamide, serum TT and FT levels decreased (P<0.01), behavior changes were basically similar to PADAM (by tail suspension test), indicating a successful modeling. After treatment serum rTT and FT levels in the testosterone propionate group and the TCZYP group significantly increased when compared with before treatment and with the model group after treatment (P<0.01). There was no statistical difference between the TCZYP group and the testosterone propionate group (P>0.05). The immobility time in the tail suspension test were significantly shortened in the testosterone propionate group and the TCZYP group (P<0.05). The exhausted swimming time was more significantly prolongated than that of the model group (P<0.05). The stAR protein gray value significantly decreased (P<0. 01). CONCLUSION: TCZYP could prevent serum TT and FT levels from decrease, improve stAR protein activities, and attenuate the depression state and muscular tension in PADAM rats. Its action was equivalent to that of testosterone propionate.

[Effects of zhibal dihuang decotion on UU-infected rat's spermatogenic cell apoptosis and expressions of caspase-3 and caspase-9].

OBJECTIVE: To observe the effects of Zhibai Dihuang Decotion (ZDD) on the ureaplasma urealyticum (UU)-infected rats' spermatogenic cell apoptosis and expressions of Caspase-3 and Caspase-9. METHODS: 45 out of 60 male SD rats were randomly selected and made into the UU infected animal model. The rest 15 were taken as the sham-operation group. The UU infected model animals were then randomly divided into the model group, the minocycline group, and the ZDD group. From the 10th day after inoculation, normal saline was given to rats of the model group and the sham-operation group by gastrogavage, while corresponding medicines were given to rats in the minocycline group and the ZDD group. All rats were killed after 21 successive days of gastrogavage. The apoptosis rate of reproductive cells, Caspase-3 and Caspase-9 expression levels and ultrastructure changes of spermatogenic cells of each group were detected and compared. RESULTS: There was statistical difference in the positive rate of the UU cultivation results, the apoptosis rate of reproductive cells, Caspase-3 and Caspase-9 expression levels in the sham-operation group, the minocycline group, and the ZDD group when compared with the model group (P<0.05). There was statistical difference in the aforesaid indices in the minocycline group and the ZDD group when compared with the sham-operation group (P<0.05). Still there was no statistical difference in the aforesaid indices between the minocycline group and the ZDD group (P>0.05). CONCLUSIONS: UU infection can lead to the increasing of spermatogenic cell's apoptosis in rats. ZDD could actively inhibit the growth and production of UU with anti-UU. One of the mechanisms of ZDD in treating UU infection and improving the sperm quality is through regulating the expressions of the apoptosis effect factors Caspase-3 and Caspase-9.

Exploring the Mechanism of Zhibai Dihuang Decoction in the Treatment of Ureaplasma Urealyticum-Induced Orchitis Based on Integrated Pharmacology.

Background: Ureaplasma urealyticum (UU) infection is the most common cause of male infertility. Zhibai Dihuang Decoction (ZBDHD) can improve the rate of forwarding motility sperm, sperm deformity rate, seminal plasma zinc and refined berry sugar levels. Methods: The potential targets of ZBDHD are obtained from The Encyclopedia of Traditional Chinese Medicine (ETCM). Orchitis-related targets were collected from the Genecards and OMIM databases. The Cytoscape and the Database for Annotation, Visualization and Integrated Discovery (DAVID) were utilized to construct and analyzed the networks. Finally, a rat model of orchitis caused by UU infection was used to detect related indicators of mitochondrial energy metabolism using TUNEL apoptosis detection technology, loss cytometry, Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) and Western Blot. Results: A total of 795 ZBDHD targets and 242 orchitis-related targets were obtained. The "ZBDHD- orchitis PPI network" was constructed and analyzed. ZBDHD can regulate signaling pathways and biological processes related to mitochondrial energy metabolism. The results of experimental studies have shown that ZBDHD maintains the integrity of sperm mitochondrial respiratory chain function by enhancing mitochondrial Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities, promotes the synthesis of mitochondrial ATP, and improves sperm energy supply, thereby improving the motility, vitality and survival rate of sperm, and effectively improving the quality of semen in UU-infected rats (p < 0.05). Conclusion:This study discovered the multi-pathway mechanism of ZBDHD intervention in UU-induced orchitis through integrated pharmacological strategies, which provides a reference for further research on the mechanism of ZBDHD intervention in orchitis in the direction of mitochondrial energy metabolism.