Mechanism of Zhibai Dihuang decotion in treating postmenopausal osteoporosis / 药学学报

In this study, the ovarian surgery (ovariectomy, OVX) was used to establish the osteoporosis mice model of primary menstruation, in order to evaluate the protective effects and mechanisms of Zhibai Dihuang decotion on postmenopausal osteoporosis (PMOP). The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Jinan University (number: 20210315-03), in compliance with the Institutional Animal Care Guidelines. C57BL/6 mice were divided into five groups, including Sham group, OVX group, low (32 g·kg-1·day-1) and high dose (64 g·kg-1·day-1) of Zhibai Dihuang decotion groups, positive drug group (alendronate, 9.9 mg·kg-1·q3d). After modeling, mice were given medication intervention for 8 weeks, and then femoral and tibial tissues were taken to detect indicators such as bone microstructure, bone resorption, and oxidative stress. The experimental results showed that after Zhibai Dihuang decotion administration, the bone microstructure damage caused by OVX surgery was alleviated, and the relevant parameters bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular number (Tb. N) and connectivity density (Conn. D) both significantly increased. At the same time, the number of TRAP positive osteoclasts decreased significantly, and the levels of proteins and genes related to osteoclast differentiation decreased, indicating that Zhibai Dihuang decoction could inhibit the increased activity of osteoclast caused by OVX. Afterwards, network pharmacology was used to construct the active compound action target network of Zhibai Dihuang decotion, and it was found that the target genes of its active ingredients were closely related to the oxidative stress pathway. Finally, the detection results of oxidative stress levels in bone tissues showed that after treatment with Zhibai Dihuang decotion, the levels of oxidative stress products 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA) in bone tissues of mice significantly decreased, while the levels of antioxidant stress substance L-glutathione (GSH) increased. These above results indicated that Zhibai Dihuang decotion can regulate the level of oxidative stress in the body and inhibit osteoclast activity, which played a therapeutic role in PMOP, as well as provided theoretical basis for the prevention and treatment of PMOP with traditional Chinese medicine.

[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.

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.

[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.

Zhibai Dihuang Decoction improves sperm mitochondrial permeability transition in rats with ureaplasma urealyticum infection / 中华男科学杂志

<p><b>Objective</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>

[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.

Effects of Zhibai Dihuang Decoction on mitochondrial cytochrome oxidase in the spermatogenic cells of rats with ureaplasma urealyticum infection / 中华男科学杂志

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 (［1.03 ± 0.09］ vs ［0.07 ± 0.03］ %, P0.05), average path velocity (VAP) (［16.22 ± 1.52］ vs ［10.05 ± 1.80］ μm/s, P0.05), 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.

[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.

Clinical Study onZhibai Dihuang Decoction Combined with Endocrine Therapy in Treating Kidney-yin Deficiency Type of Advanced Prostate Cancer / 中国中医药信息杂志

Objective To observe the clinical efficacy ofZhibai Dihuang Decoction combined with endocrine therapy for kidney-yin deficiency type of advanced prostate cancer and its influence on patient symptoms of hot flashes.Methods Totally 72 cases of advanced prostate cancer patients were randomly divided into the combined group (36 cases) and the control group (36 cases). Patients in the control group were treated with endocrine therapy, while patients in the combined group were additionally treated byZhibai Dihuang Decoction. After 12 weeks of treatment, serum prostate specific antigen (PSA), the international prostate symptom score (I-PSS), the clinical symptoms score, the number of cases of hot flashes and safety index before and after treatment were observed. Results In the course of treatment, the control group and the combined group each had 1 case lost to follow up. PSA and I-PSS in the two groups significantly decreased after treatment (P<0.01). The total efficiency rate of TCM syndrome was 94.3% (33/35) in the combined group, while it was 57.1% (20/35) in the control group, with significant difference (P<0.01). After treatment, the incidence of hot flushes was reduced after treatment in the combined group, but the control group increased, with significant difference (P<0.01). No adverse reaction was found in the two groups.ConclusionZhibai Dihuang Decoction combined with endocrine therapy is more effective than endocrine therapy alone, which can reduce the symptoms of hot flashes.

Zhibai Dihuang Decoction improves the activity of sperm mitochondrial respiratory chain complex in rats with Ureaplasma urealyticum infection / 中华男科学杂志

<p><b>Objective</b>To investigate the effect of Zhibai Dihuang Decoction (ZDD) on the sperm mitochondrial respiratory chain complex (MRCC) in rats with Ureaplasma urealyticum (UU) infection.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>