ABSTRACT
ObjectiveTo explore the effects and possible mechanisms of Jianpi Bushen Formula （健脾补肾方） on radiation-induced immune function damage of mice. MethodsFifty mice were randomly divided into four groups： normal group， model group， thymosin group， high- and low-dose groups of Jianpi Bushen Formula， with 10 mice in each group. Except for the normal group， the mice in the other groups were irradiated with a single whole-body dose of 6.0 Gy X-rays to establish a radiation-injured mouse model. After the successful modeling， the low- and high-dose groups of the Jianpi Bushen Formula were given respectively 13 g/（kg·d）、 26 g/（kg·d） of the formula by gavage， while the thymosin group was given 11.7 mg/（kg·d） of thymosin by gavage， and the normal group and model group were given 0.1 ml/（10g·d） of 0.9% sodium chloride solution by gavage. Each group was administered once a day for 7 consecutive days. After the last gavage， the mice were weighed， and their spleens were separated and weighed to calculate the spleen index. The levels of interferon gamma （IFN-γ） and interleukin 2 （IL-2） in the spleen tissue were detected by enzyme linked immunosorbent assay （ELISA）. The autophagosomes in the spleen were observed by transmission electron microscopy. The mRNA expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， and mammalian target of rapamycin （mTOR） in the spleen were detected by real-time fluorescent quantitative PCR. The protein expression of PI3K， Akt， and mTOR in the spleen， as well as the expression of autophagy-related proteins microtubule-associated light chain protein 3 （LC3）， Beclin1， and p62 were detected by Western blot. ResultsCompared with the normal group， the model group showed significant decreases in body weight， spleen index， and levels of IFN-γ and IL-2 in the spleen （P＜0.01）； the mRNA and protein expression levels of PI3K， Akt， and mTOR in the spleen were also significantly reduced （P＜0.01）； the expression of Beclin1 protein， the ratio of LC3-II/LC3-I significantly increased （P＜0.01）， while the level of p62 protein expression significantly decreased （P＜0.01）. And transmission electron microscopy showed a significant increase in the number of autophagosomes in the spleen and severe cell structure damage in the model group. Compared with the model group， all the above indicators in each medication group were significantly improved （P＜0.05 or P＜0.01）. In the high-dose Jianpi Bushen Formula group， partial intact cristae were visible in the fine mitochondria of the spleen， and there were more autophagosomes. In the low-dose Jianpi Bushen Formula group and thymosin group， the structure of the fine mitochondria in the spleen was relatively intact， and there were fewer autophagosomes. The improvement effect of the low-dose Jianpi Bushen Formula group was better than that of the high-dose group （P＜0.05 or P＜0.01）， and there was no significant difference between the low-dose group and the thymosin group in terms of each indicator （P＞0.05）. ConclusionJianpi Bushen Formula may alleviate the structural damage of the spleen， promote the recovery of immune function， and achieve a best effect at a low dose by enhancing the PI3K/Akt/mTOR signaling pathway in the spleen and inhibiting the over-activation of autophagy induced by radiation.
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Objective To investigate the mechanism of Chinese herbal medicine compound Tang Mai Ping (TMP) capsule to type 2 diabet in rats. Methods The models of type 2 diabetic rats were established by injecting lower dosage of streptozotocin (STZ, 35 mg/kg) to belly, and fed high sucrose and high fat diet. Modle rats were randomly divided into model group, TMP group, and Erjiashuanggu (EJS) group, at the same time normal control. The content of plasma insulin was detected by radio-immunicy, mRNA expressing of frame muscle of Glucose Transporter 4 (GluT4) gene was detected with hyvirdism in situ. Results After built model, levels of insulin were obviously ascended in rats, but the expression of GluT4 mRNA lower significantly (P
ABSTRACT
BACKGROUND: Chinese herb digan oral liquid promotes manufacture of hematopoietic cells. During irradiative injury, how are the therapeutic effects of intervention with Chinese herbal compound on expressions of stromal cellular adherent molecule (SCAM) and cyclin gene that are closely related to hematopoietic cells?OBJECTIVE: Mouse model with irradiative injury was established to assay the effects of Chinese herbal compound-digan oral lipid on the expression of medullary cyclinD1mRNA and the expressions of hematopoietic stem cells (HSCs), stromal cellular adherent molecular CD45 (SCAMCD45) and vascular cellular adherent moldecular-1 (VCAM-1).DESIGN: Randomized controlled experiment was designed.SETTING: Department of Integrated Traditional Chinese and Western Medicine, Wuhan General Hospital of Guangzhou Military Area Command of Chinese PLA; Institute of Hematology of Tongji Medical College and Institute of Hepatology of Hubei College of Traditional Chinese Medicine.MATERIALS: The experiment was performed in Institute of Hematology of Tongji Medical College and Institute of Hepatology of Hubei College of Traditional Chinese Medicine from January 2001 to June 2002, in which,48 outbred Kunming mice were employed, aged varied from 8 to 12weeks, mass-weighted varied from 18 to 22 g, of either sex. Drugs: 100% digan oral lipid is composed of shudihuang (radix rehmanniae preparata) and zhigancao (radix glycyrrhizae preparata). At ratio of 2/1, water decoction was prepared and concentrated as 100% oral liquid (containing raw herb 1 g/mL). METHODS: Totally 48 mice were randomized into 4 groups, 12 mice in each one. Normal group: no any management. Oral lipid group: Digan oral lipid was applied for gastric perfusion in model with irradiative injury,0.02 ml/g, twice a day. Model group: After modeling, physiological saline was used for gastric perfusion with the same dosage as the above. Batilol group: After modeling, batilol suspension was used for gastric perfusion (0.1 g/kg). Gastric perfusion was done continuously for 9 days in each group. On the 10th day, hybridization in situ was used to assay cyclinD1 mRNA expression of medullary mononuclear cell (MNC), Immuniohisto chemical SABC method was used to assay medullary VCAM-1 expression and flow cytometry (FCM) was used to assay CD45 content.MAIN OUTCOME MEASURES: CyclinD1 mRNA expression of medullary mononuclear cell (MNC) and expression of medullary stromal cellular VCAM-1 and CD45 antigenRESULTS: After modeling, there were 12 mice in normal group and 10 mice in rest groups. In result analysis, 5 mice were randomized in each group. ① In oral liquid group, the expression of medullary cellular cyclinD1 mRNA and VCAM-1 were higher significantly than model group[oralliquid group: (27.34±4.49), (53.94±8.67)% and model group: (16.06±4.57),(31.92±5.01)%, P＜ 0.01]. ② Expression of CD45 antigen iroral liquid group was higher significantly than model group [(41.38±5.76), (19.24±4.98)%, P ＜ 0.01]. Both digan oral liquid and batilol improved the expressions of cyclinD1,VCAM-1 and CD45 antigen.CONCLUSION: Chinese herbal compound-digan oral liquid enhances the expression of adherent molecule of hematopoietic stem cells, promotes the expression of stroma] cellular adherent molecule and improves the transcription of medullary cellular cyclinD1, which induces hematopoietic cells to be differentiated more maturely and to be transported peripherally so as to supplement blood.