1.Study on the extraction process of Hongqijin granules based on standard decoction
Lingrui QIN ; Xutong WU ; Yicheng SUN ; Kaiping ZOU ; Liyuan TIAN ; Xiying TAN ; Jingxian XUE ; Chenguang ZHAO ; Shun LIU
China Pharmacy 2026;37(15):1954-1959
OBJECTIVE To study the extraction process of Hongqijin granules with the standard decoction of Hongqijin formula as the reference. METHODS Fifteen batches of Hongqijin formula standard decoction were prepared. The reference characteristic chromatogram was established, and the chromatographic similarity was calculated. The average extraction yield and average transfer rates of calycosin-7-glucoside, salidroside, curcumin, and emodin of the 15 batches of standard decoction were determined. Taking soaking time, water addition amount, and decoction time as influencing factors, and extraction yield, transfer rates of marker components, as well as chromatographic similarity with the reference characteristic chromatogram as evaluation indexes, orthogonal test combined with principal component analysis was adopted to optimize the extraction process of Hongqijin granules, followed by process verification. RESULTS The average extraction yield of 15 batches of Hongqijin formula standard decoction was 32.55%. The average transfer rates of calycosin-7-glucoside, salidroside, curcumin, and emodin were 70.68%, 74.35%, 2.16%, and 4.62%, respectively. The optimal extraction process of Hongqijin granules was confirmed as follows: extraction was carried out twice; the first extraction used 8-fold water for 60 min, and the second extraction adopted 6.5-fold water for 45 min. In three validation tests, the average extraction yield was 33.01%. The average transfer rates of the four marker components were 79.51%, 75.54%, 1.84%, and 4.30%, with all RSD values less than 2%. The chromatographic similarities between the extracting solution and reference characteristic chromatogram were all higher than 0.990. CONCLUSIONS The optimized extraction process of Hongqijin granules exhibits good stability, and the material basis of the extracting solution is consistent with that of Hongqijin formula standard decoction.
2.Effect of estradiol on the gene expression of peroxisome proliferator-activated receptor gamma-2 in rat bone marrow stromal cells
Chengliang YUAN ; Xiaolian JIN ; Shigang WEI ; Xiying ZOU
Chinese Journal of Tissue Engineering Research 2007;11(3):596-600
BACKGROUND: Whehter estradiol (E2) can change the expression of peroxisome proliferator-activated receptor gamma-2(PPAR-γ2) during differentiation of bone marrow stroma cells should be studied further.OBJECTIVE: To investigate the effects of 17β-estradiol (E2) on the gene expression of PPAR-γ2 mRNA and PPAR-γ2 protein.DESIGN: Contrast observational trial.SETTING: Laboratory Department of People's Hospital of Deyang City; Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech Limited Company.MATERTALS: A 3-month-old female SD rat (200±20) g used to isolate bone marrow stromal cells was obtained from Animal center of Chengdu Chinese Medicine University (Medical Experimental Animal Number 11). Dulbecco's mimimum essential medium (DMEM) was obtained from Hyctone Compnay. 1α, 25(OH)2D3, dexamethasome (DEX) and E2 were purchased from Sigma Company. Total RNA kits were obtained from Omga. One step RNA PCR kit (AMV) was obtained from Takara Shuzo Co, Ltd. Northern direct HRP labeling and detection kit was purchased from PIERCE. Western blotting luminol reagent was obtained from Santa cruz.METHODS: The experiment was carried out from April 2001 to July 2002 in the Laboratory Department of People's Hospital of Deyang City, Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech (0, 0.1, 10, 1 000 n) was used to interfere cell differentiation for 3 days. Cultured cells were crushed with Tris-Triton X-100 PBS; activity of alkaline phosphatase was detected with Beckman CX-7 biochemical analytical device; effect of 100 g/L formalin, stained with Weigert-hematoxylin for 10 minutes, rinsed with water, differentiated with 5 g/L hydrochloric ethanol, stained with Van Gieson, desiccated with ethanol of the fractional volume of 0.95, cleared with dimethylbenzene with RT-PCR, Northern blot and Western blot during cell differentiation.protein.proliferated within 24-72 hours. Cells shaped as triangle, multiple angles and fusiform. Three days later, volume of adherent cells was increased and colony, and 10 days later, they confluenced. Bone marrow stroma cells in many generalight red to yellow). Red plasma presented synthesis of collagen. The deeper the red was, the more the collagens were.pression of PPAR-γ2 mRNA was (4.0±0.4)%, (1.7±0.2)% and (2.8±0.2)% (t=6.1, 7.2, 11.5, P< 0.01), which was higher crease the expression of PPAR-γ2 protein. When concentration of E2 was 0.1, 10 and 1 000 nmol/L, expression of PPAR-γ2 protein was (2.2±0.2)%, (2.6±0.2)% and (4.1 ±0.2)%, which was higher than that in 0 nmol/L E2 group [(1.2±0.10)%, t=6.6, 8.5, 13.2, P<0.01].CONCLUSTON: E2 can inhibit expression of alkaline phosphatase and promote differentiation of bone marrow stromal cells and expression of PPAR-γ2 mRNA and PPAR-γ2 protein.
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