ABSTRACT
Objective:To study the effect of the kidney-tonifying recipes on mouse syndrome and adrenal function induced by low-dose hydrocortisone reduction and discontinuation. Method:The 48 male ICR mice were randomly divided into normal control group, hydrocortisone group, Liuwei Dihuangtang group and Guifu Dihuangtang group, with 12 rats in each group. Each group was given 1.32 mg·kg-1·d-1 hydrocortisone for 28 days, and half a dose for 7 days, and the administration was discontinued 14 days before modeling. In the reduction and discontinuation phase, 12.5 g·kg-1·d-1 Liuwei Dihuangtang and 13.5 g·kg-1·d-1 Guifu Dihuangtang were simultaneously administered to the corresponding group for 21 d. On the 28th and 49th day of the experiment, the characteristic information of mice was detected by the experimental methodology of syndrome differentiation and treatment in mice (holding power, body surface infrared temperature). The mice were sacrificed on the 50th day of the experiment, the spleen and thymus were separated, and the spleen and thymus index were calculated, serum corticosterone content is measured by enzyme-linked immunosorbent assay (ELISA), the total RNA of the adrenal gland was extracted, and the expressions of Star, cholesterol-side-chain cleavage enzyme (Cyp11a1), Cyp21a1, Cyp11b1, low-density lipoprotein receptor (Ldlr), scavenger receptor class B type I (Scarb1/SRB1), Hmgcr, acyl-CoA-cholesterol acyltransferase-1 (Acat1), hormone sensitive lipase (Lipe/HSL), insulin-inducible gene 1 (Insig1) and sterol regulatory element binding transcription factor 2 (Srebf2) were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Western blot was used to detect the expressions of LDLR, SRB1, CYP11A1 and cytochrome P450 family 21 subfamily A member 2 (CYP21A2) in adrenal gland. Result:Compared with the normal control group, the body weight and the body mean infrared temperature of the model group were significantly decreased after 28 days of continuous administration (P<0.01). After the administration of the drug, the mice in the model group showed a significant decrease in the grip strength (P<0.05), a significant increase in the thymus index (P<0.01), a significant decrease in the serum corticosterone content (P<0.05). Expressions of adrenal Cyp21a1 and Hmgcr genes decreased (P<0.05, P<0.01), Lipe and Acat1 gene expressions increased significantly (P<0.01), adrenal CYP11A1, SRBI protein expressions decreased significantly, and LDLR protein expression increased. Compared with the model group, the body weight of Guifu Dihuangtang group decreased significantly after 21 days of treatment with traditional Chinese medicine (P<0.01), the spleen index of Liuwei Dihuangtang group decreased significantly (P<0.01), expressions of Cyp11a1, Cyp21a1, Acat1 and Lipe genes in Guifu Dihuangtang group were significantly increased (P<0.05, P<0.01), expressions of Ldlr and Scarb1 genes were significantly decreased (P<0.05), and expressions of Ldlr, Acat1 and Lipe genes in Liuwei Dihuangtang group were significantly decreased (P<0.05, P<0.01), and expressions of CYP11A1 and SRBI proteins in Liuwei Dihuangtang and Guifu Dihuangtang increased. Conclusion:Guifu Dihuangtang can promote the recovery of adrenal function in mice with drug-induced deficiency syndrome by correcting expression of adrenal cortex synthase.
ABSTRACT
Objective: To study the effects of different Chinese materia medica combinations on the regulation of adrenal function. Methods Mice with 1.65 mg/(kg•d) hydrocortisone for 14 d to induce drug-induced syndrome was treated synchronously with assissting yang and dissipating cold (Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex), nourishing yin and reducing fire (Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex), tonifying qi and promoting fluid production (Ginseng Radix et Rhizoma-Astragali Radix), and tonifying blood and benefiting spirit (Rehmanniae Radix Praeparata-Corni Fructus). The body mass of mice was dynamically observed every day, and the characteristic information such as body mass, holding power, axillary temperature, activity degrees of open field, and infrared temperature were detected by the experimental methodology of mice syndrome differentiation; The mice were sacrificed whose spleen, thymus was weighed and the organ index was calculated. Serum corticosterone, ACTH, adrenaline, and noradrenaline were measured by ELISA. The gene expressions of Star, Cyp11a1, Cyp21a1, Cyp11b1, Cyp11b2, Dbh, Ddc, Pnmt, and Th in the adrenal were detected by real-time fluorescent quantitative PCR. The protein’s expressions of LDLR, SRB1, and StAR were detected by Western blotting. Results:The effects of four different treatments on the reduction of body weight and body temperature caused by hydrocortisone were not significant. The holding power of mice in Aconm Lateralis Radix Praeparaia-Cinnamomi Cortex group was significantly higher than that of hydrocortisone group (P < 0.05). Compared with the hydrocortisone group, the serum corticosterone content of mice in Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex group increased significantly (P < 0.01); The genes expression of Star, Cyp11b1, and Cyp11b2 was significantly up-regulated (P < 0.05, 0.01); The proteins expression of SRBI and StAR were significantly up-regulated. Conclusion: The nourishing yin and purging fire treatment group (Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex) is the best to correct and protect the adrenal cortex function in mice with hydrocortisone induced syndrome at low dose.