| Metallothionein(MT)is a class of low molecular weight metal binding proteins which is rich in of cysteine.It has many biological functions,such as antioxidation,radioresistance,detoxicate,and so on.At present,the commercialized MT is mainly produced by inducing metal zinc to animal livers.Shortcomings such as sophisticated technology,low yield and high production costs greatly limit the development and application of MT.The Metallothionein-like proteins induced in C.vugularis are more conducive to industrial production with features like short extraction cycle,convenient production technology and low cost.Based on this,this article studied detoxification tests of MT on cadmium poisoning mice.And we developed two skincare products to investigate its antioxidant properties.The main research work is described as follows:The first part presented the research on detoxification with MT in cadmium poisoning mice.According to Bliss method,the LD50 value of CdCl2 for acute toxicity in mice was 11.5 mg/kg·bw.A chronic cadmium poisoning model of mice was established by continuous intraperitoneal injection of 1 mg/kg·bw CdCl2 for 7 days.Acute cadmium poisoning mice model were established by intraperitoneal injection of 5 mg/kg·bw CdCl2.After the intraperitoneal injection,we detoxified those mice with different concentrations of metallothionein.The results showed that after treated with 1 mg/kg of MT-like for 14 days,the liver cadmium removal rate reached 55.07%and the kidney cadmium removal rate reached 64.89%in the chronic cadmium poisoning model.In the acute cadmium poisoning mice,the cadmium removal rate was 66.72%and the kidney removal rate was 59.30%.In the meantime,the cadmium content in the organs was significantly reduced and showed a dose-dependent relationship.The pathological slices of mice liver showed that cadmium had obviouslys damaged to the liver tissue of mice,and the structure of mouse liver cells basically recovered after the treatment.The result showed that the MT-like could effectively promote the cadmium excretion and the gradual recovery of the damaged tissues.The second part is the application of MT-like in skincare products.The antioxidation and anti UV activity of metallothionein were investigated in the research and two products of skin care water and skin cream were developed.The physicochemical and skin irritation indexes and the microbiological experiments results showed that both two products were safe and non-irritating.After the proliferation activity of HaCat cells and the investigation of tyrosinase activity and melanin content in B16 cells,it was found that the MT-like skincare water has almost no toxicity to HaCat cells.The MT-like skincare water can also significantly inhibit tyrosinase activity in B16 cells and it can reduce the content of melanin(p<0.05).The MT-like skin cream can significantly inhibit the tyrosinase activity of B16 cells and reduce the melanin content of B16 cells.However,there is no correlation between those characteristics and the MT-like dose.In addition,it was found that MT-like skincare water can effectively restored the cell proliferation activity of HSF irradiated by UVA,and significantly inhibited the increase of ROS content in HSF cells.However,the MT-like skincream significantly lower the cell proliferation activity of the HSF cells after UVA irradiation,which has not significantly reduced of the ROS level.Therefore,the MT-like skincare water has good antioxidant capacity,and is expected to be further studied and developed. |
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