ABSTRACT
ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.
ABSTRACT
ObjectiveTo observe the effect of Hedysari Radix polysaccharide （HRP） on the Janus kinase 2 （JAK2）/signal transducer and activator of transcription protein 3 （STAT3） signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group， irbesartan group （irbesartan suspension， 22.75 mg·kg-1）， and high-， medium-， and low-dose HRP groups （HRP suspension， 200， 100， 50 mg·kg-1） according to the body weight， with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage， while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid （UA）， triglycerides （TG）， and total cholesterol （TC） were detected. Periodic acid-Schiff （PAS） staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and mRNA expression levels of JAK2， STAT3， suppressor of cytokine signaling 3 （SOCS3）， and tumor necrosis factor-α （TNF-α） in the kidney. ResultAfter 12 weeks of treatment， compared with the normal group， the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA， TG， and TC levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA， TG， and TC levels （P<0.05， P<0.01）. Compared with the normal group， the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.05， P<0.01）. ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent， and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.
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Objective: To investigate the protective effect of Hedysari Radix polysaccharide (HRP) on liver fibrosis in mice and the correlation between the hepatic fibrosis and bone loss. Methods: The mice were sc injected with 40% carbon tetrachloride (CCl4) dissolved in olive oil solution (0.1 mL/kg, once every 5 days ), while given high-, mid-, and low-dose (20, 10, and 5 g crude drug/kg) HRP and colchicines (0.6 mg/kg) daily. Continuous 35 d later, the activity of alanine transarninase (ALT) and aspartate transferase (AST), contents of total protein (TP) and albumin (ALB) were investigated by related reagent kit, and then, calculated the albumin/globulin (A/G) ratio, liver and thymus indexes and liver histological of tissue pathology were determined. The serum level of acid phosphatase resistant to acid phosphatase (TRACP), alkaline phosphatase (ALP), Ca, P, and the bone level of hydroxyproline (Hyp) were investigated by related reagent kit to observe bone loss. Results: The model group compared with the Sham group, the levels of ALT, AST, and TP increase, showed obvious liver pathological damage, liver and thymus index increase, levels of TRACP, ALP, Ca, and P decrease, and Hyp increase. HRP could significantly improve the hepatic fibrosis and bone loss induced by CCl4 in mice. Conclusion: The liver fibrosis in mice could also cause bone loss and be the primary cause of bone loss in this experiment. The certain market value of this Chinese materia medica is developed for its significant effect and contact between hepatic fibrosis and bone loss is initially investigated, which lays the foundation to further discuss the mechanism of the above two.
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Objective To investigate the improving effect of Hedysari radix polysaccharide on the behavior and its influence on cerebral central neurotransmitters in senescence accelerated mouse prone 8 (SAMP 8).Methods The senescence accelerated mice 8 were randomly divided into a model group,a Hedysari radix polysaccharide (HRP) group and a aricept group.SAMRl were served as the control group.After intragastrical treatment for 3 months,the behavior changes and the levels of norepinephrine (NE),dopamine (DA),5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were observed respectively by applying Morris water maze and high performance liquid chromatography (HPLC).Results In the SAMP8 model group,the hidden platform test showed that the latency to find the hidden platform was remarkably prolonged,the latency range from third day to fifth day was (55.31 ±9.62) s,(58.67± 10.89) s,(58.45± 10.53) s,respectively.The level ofNE、DA、5-HT and 5-HIAA was (4.97± 1.74) mmol/L,(1.63±0.57)mmol/L,(4.29±0.94)mmol/L,(2.91±0.85)mmol/L respectively,significantly reduced (P＜0.01).Compared with model group,the latency of hidden platform test was (44.73 ± 9.83) s,(40.53±8.37)s,(44.75±9.16)s respectively,which were remarkably shortened.The expression level ofNE,DA,5-HT and 5-HIAA in the HRP treatment group was (7.58± 1.62) mmol/L,(3.25 ±0.70) mmol/L,(7.04±0.95) mmol/L,(4.62±0.79) mmol/L respectively,which obviously increased (P＜ 0.05).Conclusion HRP could improve learning and memory ability of SAMP8 and the effect may related to up-regulate the expression level of cerebral Central neurotransmitters.