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Objective:To explore the effect of Gubi Decoction on serum related inflammatory factors and PI3K/Akt/mTOR signaling pathway in osteoarthritis model rats. Methods:Seventy SPF rats were randomly divided into the blank group, sham operation group, Glucosamine sulfate group, and the low, medium and high dose Gubi Decoction groups. Except the blank group and sham operation group, knee osteoarthritis animal models were prepared by the modified Hulth method in each group. On the 28th day after successful model preparation, the high, medium and low dose Gubi Decoction groups were given Gubi Decoction 24, 12 and 6 g/kg by gavage respectively; glucosamine sulfate group was given glucosamine sulfate tablet suspension 3 g/L by gavage, once a day for 28 days. The levels of TNF-α and IL-1β in serum were detected by ELISA. The gene expressions of PI3K, Akt and mTOR in cartilage tissue were detected by Real-PCR. The protein expressions of PI3K, Akt, p-PI3K, p-Akt and mTOR were detected by Western blot. Results:Compared with the model group, the knee joint diameter[(11.17 ± 1.81) mm, (11.60 ± 1.38) mm, (10.80 ± 1.17) mm vs. (12.57 ± 0.98) mm] of the rats in the glucosamine sulfate group and the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The content of TNF-α [(111.43 ± 21.98) ng/L, (53.42 ± 13.25) ng/L vs. (157.89 ± 23.60) ng/L], IL-1β [(67.50 ± 18.44) ng/L, (48.22 ± 9.63) ng/L vs. (96.11 ± 14.85) ng/L] in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of PI3K (1.87 ± 0.17, 1.24 ± 0.49 vs. 2.19 ± 0.47), Akt (1.50 ± 0.51, 1.10 ± 0.32 vs. 2.68 ± 0.63), and mTOR (1.32 ± 0.54, 1.10 ± 0.33 vs. 2.94 ± 0.55) mRNA in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The expression of PI3K, Akt, p-PI3K, p-Akt in the low, medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of mTOR in the medium dose Gubi Decoction group significantly decreased ( P<0.05). Conclusion:Gubi Decoction can significantly reduce the level of inflammatory factors in the serum of osteoarthritis model rats, and its anti-inflammatory and analgesic effects may be related to PI3K/Akt/mTOR signaling pathway.
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Objective: To optimize the flash extraction process of the active ingredients from Lonicera japonica by Box-Behnken design and response surface methodology.Methods: Flash extraction was used.With the concentration of ethanol,solid-liquid ratio and extraction time as the main influencing factors and the overall normalization value of the transfer rates of chorogenic acid and luteoloside as the evaluation index, Box-behnken design was performed to screen out the optimal extraction conditions.The mathematics relationship between the overall normalization value and the independent variables was established by multiple linear regression and binomial fitting, and response surface methodology was used to predict the optimal process conditions.Results: The optimal extraction conditions were determined as follows: the ethathol concentration was 66.92% ,the solid-liquid ratio was 18.82 ,the extraction duration was 1.20 min and extracted only once.Under the above conditions, the extraction rate of chloragenic acid and total luteoloside was 92.87% and 87.55% , respectively.Conclusion: The optimized technology is simple,economic and practical,which can provide new ideas for the rapid extraction and quality control of active ingredients from Lonicera japonica .
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Aim To establish depression rat model with kidney-yang deficiency and evaluate the related parameters.Methods A total of 24 adult SD rats were divided into control group,mod-el group and self-healing group by body weight.Model group and self-healing group were injected hydrocortisone for 21 days, meanwhile the control group was injected sodium chloride.After modeling,the related indexs detection were taken out:open-field test,sucrose preference test and forced swim test.Then the orbital blood was taken to detect 5-HT,DA and NE by LC-MS/MS.And liver,kidney,spleen,thymus,adrenal gland,testis and epididymis were taken to calculate organ index.Results Compared with the control group,model rats displayed depres-sion and kidney yang deficiency syndrome such as dry hairs, chills,back arched and so on. Correspondingly, the body weight increased slowly.Immobility in FST was significantly in-creased. Sucrose preference, horizontal scores and vertical scores was significantly decreased.Organ index decreased in liv-er,kidney,spleen,adrenal,testis and epididymis.The level of 5-HT and NE was significantly high.Compared with the model group,5-HT,NE and DA were significantly reduced,5-HT and DA were even lower than those of the control group.Conclusion Depression rat model with kidney-yang deficiency could be in-duced by continuous injection of hydrocortisone.
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Clinically, kidney?yang deficiency is one of the main TCM syndromes in patients with major depressive disorder. Warming yang and reinforcing kidney as a therapeutic measure for the treatment of depressive disorder has a?chieved good clinical curative effect. However, there are some problems on the study of kidney?yang deficiency depression. One of the reasons is that the establishment methods of depression animal model with kidney?yang deficiency are not uni?fied. In order to provide reference and further improve the replication method of depression animal models with kidney?yang deficiency, we systematically collected and analyzed the experiment results published in the past 15 years about the depres?sion animal model with kidney?yang deficiency induced by glucocorticoid hormone drugs and find out their similarities and differences.