ABSTRACT
Objective:To explore the effect of different extracts of Thlaspi Herba on the gut microbiota of hyperuricemia mice， and to reveal the substance basis and mechanism of its hypouricemic activity. Method:Eighty-eight male Kunming mice were divided into 11 groups， including blank group， model group， allopurinol group， high and low dose groups of petroleum ether extract， high and low dose groups of ethyl acetate extract， high and low dose groups of <italic>n</italic>-butanol extract， high and low dose groups of total flavonoids extract. Mice in the blank group were given 0.5% sodium carboxymethylcellulose by gavage， and the other groups were given oteracil potassium （500 mg·kg<sup>-1</sup>） by gavage to duplicate the hyperuricemia model. After modeling for several hours， the blank group and the model group were given distilled water by gavage， while mice in the allopurinol group were given allopurinol suspension （50 mg·kg<sup>-1</sup>）， and mice in each treatment group were given high and low doses of corresponding extract （5， 2.5 g·kg<sup>-1</sup>）. The serum uric acid （SUA） level and xanthine oxidase （XOD） activity were measured after 14 days. Fresh feces were collected for 16S rDNA sequencing. Result:Compared with the blank group， SUA level and XOD activity of model group were significantly increased （<italic>P</italic><0.05）. Compared with the model group， SUA level and XOD activity of the allopurinol group were significantly decreased （<italic>P</italic><0.01）. After intervention， SUA level were significantly decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， except for high dose and low dose groups of petroleum ether extract and low dose group of total flavonoids extract， XOD activity was significantly inhibited in low dose group of petroleum ether extract， high dose group of total flavonoids extract， and high and low dose groups of <italic>n</italic>-butanol extract （<italic>P</italic><0.05， <italic>P</italic><0.01）. The high dose group of total flavonoids extract was the most significant. The results of flora sequencing showed that <italic>α</italic> diversity and abundance of the model group changed significantly， and Bacteroidetes， Firmicutes and Lactobacillaceae were significantly correlated with XOD activity. After intervention， the operational taxonomic unit （OTU）， ACE， Chao1 and Shannon indexes of the high and low dose groups of total flavonoids extract were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Relative abundance of Bacteroidetes in low dose group of ethyl acetate extract， high dose group of total flavonoids extract， and high and low dose groups of <italic>n</italic>-butanol extract was significantly decreased （<italic>P</italic><0.01）， and the relative abundance of Firmicutes was significantly increased （<italic>P</italic><0.01）. The relative abundance of Lactobacillaceae in low dose group of <italic>n</italic>-butanol extract and high dose group of total flavonoids extract was significantly increased （<italic>P</italic><0.01）. Conclusion:The effective part of Thlaspi Herba for reducing uric acid is mainly flavonoids， the improvement of SUA level and XOD activity by affecting gut microbiota such as Lactobacillaceae， Bacteroidetes and Firmicutes， may be one of its mechanisms.
ABSTRACT
Objective:To investigate the mechanism of Three Handing-Three Points on pain function in sciatic nerve injury rats by observing the changes of chemokine (C-X3-C motif) ligand 1, CX3CL1)/chemokine (C-X3-C motif) receptor 1 (CX3CR1) protein and mRNA expression in spinal dorsal horn. Methods:A total of 74 male Sprague-Dawley rats were randomly divided into normal group (n= 12), sham group (n = 24), model group (n = 25), and Three Handing-Three Points group (Tuina group,n = 13). The model group and Tuina group prepared the sciatic nerve injury model. The sham group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with Tuina manipulation emulator. The photothermal pain threshold was measured seven days after modeling and after 20 days of intervention; cumulative pain score was measured seven days after modeling, and after ten days and 20 days of intervention. The spinal dorsal horn tissues were extracted to detect the protein and mRNA expression of CX3CL1/CX3CR1 with Western blotting and RT-PCR seven days after modeling and after 20 days of intervention. The microglia morphology in spinal dorsal horn was observed with immunofluorescence after 20 days of intervention. Results:Seven days after modeling, compared with the normal group, the photothermal pain tolerance threshold increased in the model group and the sham group (P < 0.05); compared with the sham group, the cumulative pain score increased in the model group and Tuina group (P < 0.05). After ten days of intervention, the cumulative pain score was lower in Tuina group than in the model group (P < 0.05). After 20 days of intervention, both the photothermal pain tolerance threshold and cumulative pain score were lower in Tuina group than in the model group (P < 0.05). There was no significant difference in the expression of CX3CL1/CX3CR1 protein and mRNA on the seven days after modeling and after 20 days of intervention (P > 0.05). The microglia in the model group were partially activated or completely activated, while those in Tuina group were unactivated or partially activated after 20 days of intervention. Conclusion:Three Handing-Three Points can improve the pain function of sciatic nerve injured rats, which may associate with regulating microglia through the pathway other than CX3CL1/CX3CR1.
ABSTRACT
Objective:To investigate the effect of Tuina of Three Handing-Three Points on the motor function of hind limbs, the proliferation of Schwann cell, recovery of myelin sheath and the expression of transforming growth factor (TGF)-β1/Smad2 pathway protein in injured sciatic nerve of rats. Methods:A total of 66 male Sprague-Dawley rats were randomly divided into sham operation group (n = 22), model group (n = 22) and observation group (n = 22). The sciatic nerve injury model was made by clamping method. On the eighth day after modeling, the observation group received stimulation on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34). The sciatic functional index (SFI) was measured before intervention and 21 days after intervention. The Oblique Plate Test was performed before intervention, and seven days, 14 days and 21 days after intervention. The expression of S100, TGF-β1 and Smad2 were observed by immunofluorescence. The expression of TGF-β1, Smad2 and p-Smad2 was detected by Western blotting. Results:Before intervention, SFI was lower in the model group and observation group than in the sham operation group (P < 0.05); 21 days after intervention, SFI and the angle of Oblique Plate Test were higher in the observation group than in the model group (P < 0.05). Immunofluorescence showed that, 21 days after intervention, the expression of S100 was lower in the model group than in the sham operation group (P < 0.01), and was higher in the observation group than in the model group (P < 0.05), no difference was found between the observation group and the sham operation group (P > 0.05). Western blotting showed that, before intervention and seven days after intervention, the expression of TGF-β1, Smad2 and p-Smad2 were higher in the model group than in the sham operation group; 21 days after intervention, no difference was found in the expression among groups (P > 0.05) Conclusion:Three Handing-Three Points could promote the proliferation of Schwann cell and the recovery of myelin sheath, to improve the motor function of hind limbs in rats with sciatic nerve injury, which may not be related to TGF-β1/Smad2 pathway.
ABSTRACT
Objective:To explore the effect of Tuina of Three Handing-Three Points on the recovery of motor function, the expression of neuregulin (NRG) 1 and human epidermal growth factor receptor (ErbB) 2 in the injured point of sciatic nerve and L4-6 spinal cord, and the morphological change of myelin sheath at the injured point of sciatic nerve of rats. Methods:A total of 76 male Sprague-Dawley rats were randomly divided into normal group, sham operation group, model group and Tuina group with 19 rats in each group. The right side sciatic nerve was clamped to make model in the model group and Tuina group. The sham operation group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with dialing, plucking and kneading using Tuina technique simulator. All of them were tested with Oblique Plate Test before modeling, seven days and 28 days after modeling. Western blotting was used to detect the protein expression of NRG1 and ErbB2 in the injured point of sciatic nerve and L4-6 spinal cord. The change of myelin sheath at the sciatic nerve injury point was observed and analyzed by transmission electron microscope. Results:Seven days and 28 days after modeling, the scores of Oblique Plate Test were lower in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the scores was higher in Tuina group than in the model group (P < 0.05). At the sciatic nerve injury point, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); seven days and 28 days after modeling, there was no significant difference in NRG1 among groups (P > 0.05); 28 days after modeling, the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05). In L4-6 spinal cord, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and sham operation group (P < 0.05); seven days after modeling, the expression of NRG1 was higher in the model group and Tuina group than in the sham operation group (P < 0.05), and the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the expression of NRG1 was higher in Tuina group than in the model group (P < 0.05), and there was no significant difference in ErbB2 among groups (P > 0.05). The electron microscope showed that, 28 days after modeling, the myelin sheath collapsed seriously in the model group; while the ultrastructure of the nerve injury point improved, and the myelin sheath of the nerve fiber was relatively intact in Tuina group; the g-ratio value was lower in the model group than in the sham operation group (P < 0.05), the g-ratio value was higher in Tuina group than in the model group (P < 0.05), and no difference was found in g-ratio value between Tuina group and sham operation group (P > 0.05). Conclusion:Three Handing-Three Points could improve the motor function of hind limbs in rats with sciatic nerve injury, which may be related to the adjustment of NRG1 and ErbB2 in the sciatic nerve and spinal cord, to maintain normal myelin sheath structure.