The positive and negative externalities of environmental regulations affects the development of high-tech industry clusters through the location of enterprises. Using the panel data of the Yangtze River Economic Belt from 2011 to 2017, empirically, the impact of environmental regulations in the Yangtze River Economic Belt and its upper, middle and lower reaches on the agglomeration of high-tech industries was analyzed. The results show that whether it is sub-regional or as a whole, there is a "U"-shaped nonlinear relationship between the formal environmental regulations of the Yangtze River Economic Belt and the agglomeration of high-tech industries. Except for the middle reaches of the Yangtze River Economic Belt, the upper and lower reaches and the whole of the Yangtze River Economic Belt are basically still in the "U" shape. The left-hand declining stage of the U"-shaped curve means that there is a reverse relationship between formal environmental regulation and the agglomeration of high-tech industries. In informal environmental regulation, the level of income is an important factor, which affects the level of agglomeration of high-tech industries. Each area should be divided into the intensity of environmental regulations according to the actual situation, in order to reach the inflection point of the "U"-shaped curve. At the same time, all regions should also pay attention to guiding consumer preferences towards green and healthy development, forcing enterprises to innovate in science and technology, and attracting high-tech industry clusters.