| Human placenta is a traditional Chinese medicine that has the effects of tonifying the kidney,benefiting essence,reinforcing blood,benefiting qi.However,there are few reports about pig placenta related research.Pigs are mammals,pig placenta is easily available,efficacy should be similar and comparable with human placenta.In this experiment,we conducted a basic study on the pharmacodynamics of pig placenta to organize pig placenta protein(PPP)extracted by freezing and thawing,prepared pig placenta protein gel(PPPG)to examine its basic properties and efficacy.Provides basis for the use of pig placenta waste.The main research contents and results are as follows:1 Extraction of pig placental protein and study on its material basisThe fresh pig placenta tissue was first crushed by mechanical comminution,and then frozen and thawed,homogenized,filtered to obtain PPP,stored as freeze-dried powder.The total protein content was measured and the components of PPP was analyzed by thin layer chromatography(TLC),sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),etc.The results showed that PPP was a light yellow transparent solution and the lyophilized powder was milky white flocculent powder.Seven spots were detected by TLC.The R/values were 0.06,0.08,0.23,0.27,0.34,0.47,0.54,respectively;the UV spectrum scan showed that the largest absorption wavelength is at 260 nm;the trace element analysis result is:PPP solution is rich in essential nutrients such as Ca,P,K,Mg,Zn,Fe and so on;infrared spectrum scan results show that There are obvious vibration peaks nearat 3400 cm-1 and 1630 cm-1;seven electrophoresis bands were obtained by SDS-PAGE,and identified by MALDI-TOF MS.In descending order of molecular weight as:78 kDa glucose-regulated protein,heat shock 70 kDa protein 8,protein disulfide-isomerase A3 precursor,prolyl 4-hydroxylase beta polypeptide,cytoskeletal beta actin partial,aldose reductase and hemoglobin subunit beta.2 Effect of porcine placental protein on DA content in miceNormal mice and D-galactose-induced aging mice were intragastrically administered with different doses of PPP(high concentration:1 g kg-1·d-1?medium concentration:0.5 g·kg-1·d-1,low Concentration:0.25 g·kg-1·d-1).After 35 days of administration,brain tissue and blood samples at the same time,The levels of dopamine(DA)in brain tissue and blood of mice in each group were detected by enzyme-linked immunosorbent assay(ELISA)and their effects on DA neurons in vivo were examined.The experimental data showed that compared with the blank control group of normal mice,the level of DA in the normal control group,high,middle and low concentrations of PPP groups were significantly increased by 58.0%(p<0.01)and 59.0%(p<0.01),37.6%(p<0.01)and 30.2%(p<0.05),respectively.The DA content in aging mice was significantly reduced.Compared with the model group DA levels in the positive control group and the high,middle,and low concentration groups of PPP were significantly increased by 95.5%(p<0.01),101.3%(p<0.01),79%(p<0.01),and 67.1%(p<0.01),respectively.Compared with the blank control group of normal mice the levels of DA in blood of positive control group and PPP high,middle and low concentration groups increased by 11.9%(p<0.05),10.5%(p<0.05),and 6.2%(p>0.05)and 3.2%(p>0.05),respectively.Compared with the model group,DA levels in blood of mice with positive control group and high,middle and low concentrations of PPP were increased by 30.1%(p<0.01),15.7%(p<0.05),8.8%(p<0.05)and 5.5%(p>0.05),respectively.3 Preparation and efficacy of porcine placental protein gel12 g of hydrochloric acid chitosan was added to 25 mL of purified water,and it was swollen and dissolved to form a gelling solution;25 mL of purified water was added to 132 g of PPP lyophilized powder to form a lysis solution;the gel solution and the lysis solution were homogenized for 2 min then 6g of glycerol was added and homogenize for 2 min to obtain a PPP gell.The viscosity,pH,total protein concentration of the PPP gel were measured and the structure of gel was observed by scanning electron microscopy(SEM).The PPP gel was smeared on D-galactose-induced aging mice skin and deep ? degree scalded rat skin.After 35 days of administration,the effect of PPP gel on epidermal morphology and thickness in skin tissues of each group was examined by HE staining.At the same time,RT-PCR was used to detect the expression of type I procollagen mRNA in skin tissue.The results showed that the prepared PPP gel was a homogeneous and delicate yellow semi-solid material with a viscosity of(4.5±0.1)Pa.s,a pH of 6.6?6.9,total protein concentration was 5%.It was easy to apply and Clean,meets gel formulation requirements.The skin tissue sections showed that PPP gel can increase the thickness of skin epidermis in the whole and wounded areas of aging and burned animals;RT-PCR showed that the expression of type I procollagen mRNA in skin tissue of PPP gel group was significantly increased(p<0.05).This experiment extracted and identified protein or peptide substances in the placenta of pigs that can enhance cell viability and human immune function,confirming that it is related to the obvious antioxidant and anti-aging effects of pig placenta extract and highlighting its application in health care and medicine.The value and application prospects are worth learning from. |
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